Genome-wide transcriptomic analysis of n-caproic acid production in Ruminococcaceae bacterium CPB6 with lactate supplementation

Genome-wide transcriptomic analysis of n-caproic acid production in Ruminococcaceae bacterium CPB6 with lactate supplementation | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research article Genome-wide transcriptomic analysis of n -caproic acid production in Ruminococcaceae bacterium CPB6 with lactate supplementation Yong Tao, Shaowen Lu, Yi Wang, Cuicui Wei, Hong Jin, Xiaohong He This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-76088/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 12 You are reading this latest preprint version Abstract Background n -Caproic acid (CA) is gaining increased attention due to its high value as a chemical feedstock. Ruminococcaceae bacterium strain CPB6 is an anaerobic mesophilic bacterium that is highly prolific in its ability to perform chain elongation of lactate to CA. However, little is known about the genome-wide transcriptional analysis of strain CPB6 for CA production triggered by the supplementation of exogenous lactate. Results In this study, 0.5% lactate was supplemented into the fermentation with Ruminococcaceae bacterium CPB6 for CA production. Compared to the control (without lactate supplementation), lactate supplementation led to earlier CA production and higher final CA titer and productivity. Transcriptional analysis was carried out using RNA-Seq for the culture with and without lactate supplementation (two groups) at growth and stationary phases, respectively. It has been indicated that 295 genes whose changes in expression patterns were substrate and/or growth dependent. These genes cover crucial functional categories. Specifically, 5 genes responsible for the reverse β-oxidation pathway, 11 genes encoding ATP-binding cassette (ABC) transporters, 6 genes encoding substrate-binding protein (SBP) and 4 genes encoding phosphotransferase system (PTS) transporters were strikingly upregulated in response to the addition of lactate. These genes would be candidates for future studies aiming at understanding the regulatory mechanism of lactate conversion into CA, as well as for the improvement of CA production in strain CPB6. Conclusions This study suggested that lactate supplementation can promote CA production by altering the expression patterns of genes involved in the essential metabolic pathways, such as central pyruvate metabolism, the reverse β-oxidation pathway, ABC and PTS transports. The findings presented herein reveal unique insights into the biomolecular effects of lactate on CA production at the gene transcriptional level. Laboratory Diagnostics n-Caproic acid Lactate Chain elongation Transcriptome RNA-Seq Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Background The increasing demand for fuels and chemicals and the scarcity of fossil resources necessitate the development of sustainable and innovative strategies for the industrial production. Organic residual streams (e.g., food waste and brewery wastewater) have great potential to be employed as feedstock for the production of biofuels and biochemicals [ 1 , 2 ]. n -Caproic acid (CA) is a medium chain fatty acid of 6-carbon, which has recently gained considerable attention due to its high value as a fuel and chemical feedstock [ 3 ]. Biosynthesis of CA can be achieved in some anaerobic bacteria (e.g., Clostridium kluyveri ) via the chain elongation pathway with ethanol as reduced substrate (electron donor) [ 4 , 5 ], in which the oxidation of ethanol provides energy and acetyl-CoA for the reverse β-oxidation pathway [ 6 ]. Many studies show that the addition of ethanol during the acidification of wastes can promote chain elongation, and lead to higher volumetric production rates and a high CA selectivity [ 7 , 8 ], indicating that ethanol is an efficient reduced substrate for CA production. In addition, researchers have explored other chemicals used as electron donors for CA production, including hydrogen [ 9 ], methanol [ 10 ], propanol [ 11 ], and D -galactitol [ 12 ]. Recently, lactate was showed to be a potential alternative to ethanol for the production of CA by reactor microbiome systems [ 13 , 14 ]. More recently, our study demonstrates that CA is efficiently produced from lactate by the pure culture of a Ruminococcaceae bacterium CPB6 isolated from a fed-batch reactor microbiome [ 15 ]. Strain CPB6 is an anaerobic mesophilic bacterium that is highly prolific in its ability to perform chain elongation of lactate to caproate, and can produce CA (C6) from lactate (as electron donor) with C2-C4 carboxylic acids as electron acceptors, and heptoic acid (C7) with C3-C5 carboxylic acids [ 16 ]. The phylogenic analysis based on 16S rRNA sequences and the whole genome showed that strain CPB6 might belong to a new clade (genus) of the family Ruminococcaceae, it is thus tentatively christened with the name Ruminococcaceae bacterium CPB6 [ 15 , 17 ]. Furthermore, we identified sets of genes correlated with the chain elongation pathway by sequencing and annotating the whole genome of the CPB6 strain. However, to date, very little information is available on genome-wide transcriptomic analysis of the CPB6 strain for CA production using lactate as the electron donor, which is essential to understand the effect of lactate on the metabolic pathway shift for CA production at the molecular level and thus elucidate proper strategies for further strain improvement. RNA-Seq is a powerful method to elucidate the metabolism and identify specific genes/enzymes for the particular interesting metabolic pathways [ 18 , 19 ]. In the present study, RNA-Seq for the transcriptomic analysis of strain CPB6 was carried out to investigate the effect of lactate supplementation on cell growth, CA production and other essential metabolisms. The identified key genes herein provide candidates for the metabolic engineering in the future in order to develop more robust strains based on CPB6 for enhanced production of CA and relevant biochemicals. Results And Discussion Growth and the production of CA As shown in Fig. 1 a, cells took approximately 18 h to grow to the stationary phase. Although the maximum OD 600 of the culture with lactate supplementation was slightly higher than that of the control (without lactate supplementation) at the stationary phase (1.25 vs 1.16), both cultures showed similar growth kinetics, indicating that lactate supplementation had little impact on cell growth. The CA production was started to be observed in the culture with lactate supplementation after 6 h of cultivation, and the CA titer continued to increase and reached 1717.2 mg L − 1 at 21 h (Fig. 1 b), while CA production was not detected until 15 h of cultivation in the control, the CA titer of which only reached 618 mg L − 1 at 21 h. In both cultures, while more significantly in the culture with lactate supplementation, the main endproduct was CA together with small amount of butyrate, suggesting that the carbon flux of acetyl-CoA may be inclined to be channelled to CA synthesis, especially in the presence of lactate. This is consistent with our previous studies [ 15 ]. In sum, compared to the control without lactate supplementation, the lactate supplementation had little effect on the cell growth, but led to earlier initiation for CA production (6 vs 15 h), higher final CA titer (1717 vs 618 mg L − 1 ) and higher CA productivity (81.8 vs 29.4 mg L − 1 h − 1 ). RNA-Seq statistics Samples were taken for RNA-Seq analysis from both the growth (12 h) and stationary (18 h) phases for both the culture with lactate supplementation and the control. For each culture, independent biological triplicates (a, b, and c) were included (Table 1 ). Therefore, a total of twelve samples were taken for cDNA libraries construction and sequencing on the Illumina HiSeq platform (Illumina, San Diego, USA). The number of raw reads generated from the sequencing for each library was from 15.7 to 23.5 million (Table S1 in Additional files). A total of 224 Mb sequence reads from 12 cDNA libraries were mapped to the genome of strain CPB6. Only those reads that mapped unambiguously to the CPB6 genome were used for further analysis. Table 1 Summary of RNA-Seq sequencing and data analysis results Sample Name L1 L2 C1 C2 a b c a b c a b c a b c Total reads 22982792 18614536 18087962 16881648 16844058 15343814 18684112 19108148 19099082 19909258 23032120 18662552 No. of read mapped 22684858 18390709 17865180 16713319 16606726 15098988 18437261 18893873 18905208 19631686 22572873 18285805 Ratio of reads mapped (%) 98.7 98.8 98.77 99 98.59 98.4 98.68 98.88 98.98 98.61 98.01 97.98 No. of unique reads mapped 22339068 17915954 17559600 16540060 16324768 14862898 18050316 18505697 18508661 19247256 22187272 17856892 No. of genes with detectable expression 1969 1968 1968 1968 1969 1968 1968 1968 1968 1968 1969 1969 Range in expression levels (TPM) 8.3–2.7 × 10 4 8.0-1.7 × 10 4 11.5–1.7 × 10 4 3.2–5.3 × 10 4 26.4–3.7 × 10 4 6.6–7.3 × 10 4 4.3-2.0 × 10 4 3.8–1.8 × 10 4 4.1–1.9 × 10 4 5.2–2.4 × 10 4 10.8–1.8 × 10 4 24.5–1.9 × 10 4 L1: cell culture with lactate supplementation from the growth phase; L2: cell culture with lactate supplementation from the stationary phase; C1: Control culture without lactate supplementation from the growth phase; C2: Control culture without lactate supplementation from the stationary phase. a, b and c represented the biological triplicate samples. Overall, out of reads derived from all the samples, 15.1 to 22.7 million reads were unambiguously mapped to the CPB6 genome, and over 98% reads were mapped (Table 1 ). A total of 1968/1969 out of 2045 protein-coding genes had detectable expression in both conditions (with or without lactate supplementation) (Table 1 ), indicating that the RNA-Seq analysis in this study achieved comprehensive coverage of the CPB6 transcriptome. The transcription levels (the number of transcripts per million, TPM) of most active protein-coding genes were in the range of 3.2 × 10 4 –7.3 × 10 4 . As illustrated in Fig. 2 , the gene expression could be classified into four levels: low (TPM 1000). The number of genes at some specific expression levels was significantly different for the two cultures (with or without lactate supplementation). For the growth phase, there were slightly more genes in the moderate and high and very high expression level in the culture with lactate supplementation than in the control, but lowly expressed genes were significantly decreased. While for the stationary phase, the culture with lactate supplementation had more genes in the moderate expression level, but fewer genes in the high and very high expression level. These results suggested that more changes in gene expression are triggered by the addition of lactate at the stationary phases. Functional annotation and classification In the transcriptome of strain CPB6, a total of 1122 expressed genes were allocated into three primary Gene Ontology (GO) categories (Fig. 3 ), including the category of biological process (601 genes), cellular component (524 genes), and molecular function (916 genes). In each category, the genes were further assigned into 28 functional groups, such as metabolic process (478 genes), cellular process (440 genes), cell part (307 genes), membrane part (297 genes), catalytic activity (654 genes), binding (561genes), and etc. The analysis of the genes based on the KEGG annotation identified a total of 1046 unigenes allocated into six primary KEGG categories including 35 subcategories (Figure. S1 in Additional files). The top 5 categories of genes were: carbohydrate metabolism, amino acid metabolism, membrane transport, translation, and metabolism of cofactors and vitamins, respectively. The top 10 enriched pathways included ABC transporters (58 genes), Amino sugar and nucleotide sugar metabolism (27 genes), Starch and sucrose metabolism (26 genes), Glycolysis / Gluconeogenesis (24 genes), Purine metabolism (40 genes), Pyrimidine metabolism (37 genes), Peptidoglycan biosynthesis, Aminoacyl-tRNA biosynthesis (28 genes), Ribosome (52 genes), and Quorum sensing (26 genes). The analysis based on the Clusters of Orthologous Groups (COGs) showed that 1785 unigenes were allocated to four primary COG categories containing 20 COG functional clusters. The top 5 annotated genes corresponding to the KEGG pathways were Replication recombination and repair (155 genes), Translation, ribosomal structure and biogenesis (142 genes), Amino acid transport and metabolism (126 genes), Carbohydrate transport and metabolism (110 genes), and Inorganic ion transport and metabolism (Figure S2 in Additional files). Differential expression of global genes The correct identification of differentially expressed genes (DEGs) between specific conditions is a key in the understanding phenotypic variation of organisms under environmental stress. As shown in Table 2 , there were only 34 differentially expressed genes (DEGs, FC ≥ 2 or ≤ 0.5 with P-value < 0.05) identified during the growth phase between two culture conditions, of which 15 genes were upregulated, and 19 genes were downregulated more than two-fold. In addition, a total of 245 DEGs were identified at the stationary phase, of which 123 genes were significantly upregulated and 122 genes were downregulated (Table S2 in Additional files). It suggested that the addition of lactate led to differences in gene expression between the two cultures (with and without lactate supplementation) during different growth phases. Table 2 The differentially expressed genes in culture with/without lactate supplementation during the growth phase No. Gene_ID Gene name Gene description TPM FC (L1/C1) P-value C1 L1 15 upregulated genes (FC ≥ 2.0); all statistically significant (P < 0.05) 1 B6259_RS06365 atoB acetyl-CoA C-acetyltransferase 1224 5204 3.45 7.9E-39 2 B6259_RS06360 crt enoyl-CoA hydratase 795 3434 3.46 2.4E-33 3 B6259_RS06355 hbd 3-hydroxybutyryl-CoA dehydrogenase 1418 6306 3.49 2.3E-27 4 B6259_RS07830 pta phosphate acetyltransferase 271 666 2.09 4.3E-24 5 B6259_RS00440 - methionine ABC transporter ATP-binding protein 51 504 5.25 6.2E-18 6 B6259_RS00450 - metal ABC transporter substrate-binding protein 30 699 5.69 5.9E-15 7 B6259_RS00445 - ABC transporter permease 27 446 5.17 6.9E-14 8 B6259_RS08190 cysK cysteine synthase A 390 7426 4.07 6.4E-10 9 B6259_RS08440 - unknown function 1048 3598 2.33 5.2E-06 10 B6259_RS06010 - hypothetical protein 21 89 2.52 8.1E-06 11 B6259_RS07140 - hypothetical protein 154 470 2.17 3.0E-05 12 B6259_RS01720 cadA cadmium-translocating P-type ATPase 22 66 2.16 3.9E-05 13 B6259_RS06870 - Hsp20/alpha crystallin family protein 315 1102 2.21 1.6E-04 14 B6259_RS00455 pepT peptidase T 37 576 2.26 2.0E-04 15 B6259_RS02585 bdh butanol dehydrogenase 82 242 2.04 3.1E-04 19 downregulated genes (FC ≤ 0.5); all statistically significant (P < 0.05) 1 B6259_RS08515 - peptide ABC transporter substrate-binding protein 98 53 0.48 1.7E-23 2 B6259_RS09280 - PTS glucose transporter subunit IIA 1200 484 0.37 5.5E-23 3 B6259_RS09735 ilvH acetolactate synthase small subunit 564 302 0.48 9.4E-19 4 B6259_RS06995 - hypothetical protein 276 43 0.20 2.7E-18 5 B6259_RS08565 - hypothetical protein 143 79 0.50 2.3E-13 6 B6259_RS07000 - sugar ABC transporter permease 113 31 0.30 9.2E-13 7 B6259_RS01525 - unknown function 2683 1010 0.37 8.4E-12 8 B6259_RS03200 - unknown function 2683 1010 0.37 8.4E-12 9 B6259_RS07010 tag glycosylase 144 46 0.34 9.9E-11 10 B6259_RS07005 - carbohydrate ABC transporter permease 90 33 0.37 1.1E-09 11 B6259_RS01865 - DUF2520 domain-containing protein 260 85 0.36 6.7E-09 12 B6259_RS01880 panD aspartate 1-decarboxylase 444 156 0.37 9.5E-09 13 B6259_RS01870 panB 3-methyl-2-oxobutanoate hydroxymethyltransferase 314 105 0.37 1.1E-08 14 B6259_RS01875 panc pantoate-beta-alanine ligase 350 115 0.37 1.2E-08 15 B6259_RS01760 - hypothetical protein 820 369 0.44 8.9E-08 16 B6259_RS02315 - basic amino acid ABC transporter substrate-binding protein 147 79 0.50 1.8E-07 17 B6259_RS00100 fruK 1-phosphofructokinase 1256 276 0.35 1.7E-06 18 B6259_RS00095 - PTS fructose transporter subunit IIC 1273 372 0.37 1.8E-06 19 B6259_RS00105 - DeoR/GlpR transcriptional regulator 1304 278 0.36 3.6E-06 L1: lactate-supplemented cells at growth phase C1: no-lactate-supplemented cells (controls) at growth phase The COG distribution of the DEGs at both the growth phase and the stationary phase was illustrated in Fig. 4 . It revealed the potential genes related to the pathways and bioprocesses for the utilization of lactate for CA production. At the growth phase, predominant number of DEGs belong to the ‘inorganic ion transport and metabolism, [P]’ group and the ‘carbohydrate transport and metabolism, [G]’ group. While at stationary phase, most upregulated genes belong to the ‘carbohydrate transport and metabolism, [G]’ group, which play important roles for the degradation and utilization of carbohydrate substrates [ 20 , 21 ]. It was worth noting that a large number of DEGs (for both upregulated and downregulated ones) fall into the ‘function unknown, [S]’ group.), meaning that their functions are unknown. This may be because CPB6 belongs to a novel species or clade ( Clostridium cluster IV) of the family Ruminococcaceae , sharing low 16S rRNA sequence similarity (< 92.6%) with the other organisms in GenBank and RDP [ 15 , 17 ]. Cluster analysis of the DEGs between the culture with lactate supplementation and the control was showed in Fig. 5 . The results showed that the gene expression of the triplicate (a, b and c) of each sample demonstrated very similar expression patterns. According to Fig. 5 , L1 (with lactate supplementation at the growth phase) cluster was most closely with C1 (without lactate supplementation at the growth phase) cluster, indicating the presence of a small amount of DEGs caused by the addition of lactate at this phase. However, the distinct difference of gene expression was observed between the two cultures at the stationary phase (L2 vs C2), Further, venn diagram showed that 295 genes whose changes in expression patterns were substrate and/or growth dependent, of which 31 genes were substrate (lactate) dependent, 228 genes were growth dependent, and 36 genes were substrate-growth dependent (Fig. 6 a). Specifically, 11 and 20 lactate-dependent genes were significantly upregulated and downregulated, as well as 98 and 130 growth-dependent genes were significantly upregulated and downregulated, respectively (Fig. 6 b). It was suggested that the differences in gene expression are stronger for stationary phase vs growth phase than for plus/minus lactate. Similar results was observed for Clostridium thermocellum , in which growth rate had stronger effects on gene expression than substrate type (insoluble cellulose vs soluble cellobiose) [ 22 ]. These DEGs were described in detail in later section. Expression of glycolysis genes An overview of the metabolic pathway in strain CPB6 and the expression levels of genes involved in key metabolic processes with their fold change (FC) were shown in Fig. 6 and Table 3 . Generally, in clostridia, glucose is converted into pyruvate via glycolysis, and the produced pyruvate is further converted into acetyl-CoA for the production of acetate and butyrate at acidogenic phase [ 23 – 25 ]. In the present study, the expression of genes for glycolysis was detected in the CPB6 transcriptome, which reinforced its genome annotation [ 17 ]. Most glycolytic genes were expressed at a relatively high level (TPM > 150) between the culture with lactate supplementation and the control, but there was no significant difference between them at the growth phase. Three glycolytic genes exhibited different expression patterns at the stationary phase. Gene encoding phosphofructokinase (PFK, B6259_RS06095) was significantly downregulated, while genes encoding glucose-1-phosphate adenylyltransferase (GlgC, B6259_RS09035) and 1, 4-alpha-glucan branching enzyme (GlgB, B6259_RS09040) were upregulated by 4.58 and 3.42-fold in the culture with lactate supplementation than in the control, respectively. GlgB and GlgC are typically associated with glycogen synthesis, why expression of these genes be affected by lactate supplementation remains unclear. Overall, the addition of lactate has little impact on the expression of glycolytic genes. Table 3 The differentially expressed genes within the important metabolic pathways in culture with/without lactate supplementation Functional description Gene_ID TPM of genes from culture with lactate supplementation a TPM of genes from the Control a RNA relative fold change (Treatment/Control) 12 h 18 h 12 h 18 h 12 h 18 h Glycolysis PTS-Glc-EIIA, PTS glucose transporter subunit IIA B6259_RS09280 484 260 1200 517 0.37 b 0.62 GlgC, glucose-1-phosphate adenylyltransferase B6259_RS09035 153 1323 236 241 0.57 4.58 c GlgB, 1,4-alpha-glucan branching enzyme B6259_RS09040 194 745 236 201 0.72 3.42 c sugar phosphate isomerase/epimerase B6259_RS06500 181 175 150 233 1.05 0.88 PGM, phosphoglucomutase B6259_RS09200 95 189 127 113 0.66 1.80 GPI, glucose-6-phosphate isomerase B6259_RS04825 2015 1789 1833 1818 0.96 1.12 PFK, phosphofructokinase B6259_RS06095 426 97 580 516 0.66 0.23 b ALDO, fructose-bisphosphate aldolase B6259_RS00415 749 402 800 891 0.83 0.57 TPI, triose-phosphate isomerase B6259_RS09105 224 229 315 493 0.65 0.58 GapA, glyceraldehyde phosphate dehydrogenase B6259_RS09050 5322 4284 4790 7732 0.98 0.70 PGK, phosphoglycerate kinase B6259_RS09100 523 524 705 1029 0.67 0.65 gpmI, 2,3-bisphosphoglycerate-independent phosphoglycerate mutase B6259_RS09110 203 200 284 469 0.66 0.55 ENO, phosphopyruvate hydratase B6259_RS04810 41 65 30 52 1.14 1.46 PK, pyruvate kinase B6259_RS02335 254 102 293 228 0.77 1.46 Central pyruvate metabolism PpdK, pyruvate phosphate dikinase B6259_RS00120 1301 823 1163 1535 0.99 0.69 Pfor, pyruvate: ferredoxin (flavodoxin) oxidoreductase B6259_RS09135 4329 4382 2044 1225 1.83 3.26 c PCK, phosphoenolpyruvate carboxykinase B6259_RS09255 368 159 554 1031 0.62 0.23 b PflD, formate C-acetyltransferase B6259_RS09900 98 188 107 471 0.83 0.54 ADH, alcohol dehydrogenase B6259_RS03100 200 116 163 159 1.07 0.84 Incomplete TCA cycle CS, citrate synthase, citrate lyase B6259_RS03360 936 187 543 642 1.42 0.39 b ACO, aconitate hydratase B6259_RS05795 227 162 153 201 1.27 0.94 IDH, isocitrate dehydrogenase B6259_RS05805 237 232 197 291 1.04 0.93 FUM, fumarate hydratase B6259_RS07270 310 186 260 437 1.04 0.49 b PCK, phosphoenolpyruvate carboxykinase B6259_RS09255 368 159 554 1031 0.62 0.23 b Hydrogen production HydE, [FeFe] hydrogenase H-cluster B6259_RS02550 113 73 174 44 1.44 2.24 c HydF, [FeFe] hydrogenase H-cluster B6259_RS09690 67 40 50 24 1.43 1.17 Lactate fermentation pathway D-ldh, D-lactate dehydrogenase B6259_RS06770 76 88 58 108 1.14 0.95 L-ldh, L-lactate dehydrogenase B6259_RS09845 79 111 119 295 0.59 0.44 b Acetate fermentation pathway PTA, phosphate acetyltransferase B6259_RS07830 666 697 271 321 2.09 c 2.23 c ACK, acetate kinase B6259_RS03430 290 297 288 233 0.88 1.41 The reverse β-oxidation pathway AtoB, acetyl-CoA C-acetyltransferase B6259_RS06365 5204 9909 1224 1077 3.45 c 6.31 c HBD, 3-hydroxybutyryl-CoA dehydrogenase B6259_RS06355 6306 13975 1418 1022 3.49 c 8.59 c CRT, enoyl-CoA hydratase B6259_RS06360 3434 7348 795 647 3.46 c 7.34 c BCD1, butyryl-CoA dehydrogenase B6259_RS01790 3278 3104 3787 3014 0.76 1.19 BCD2, butyryl-CoA dehydrogenase B6259_RS02600 42 313 41 66 0.90 4.49 c EtfA, electron transfer flavoprotein subunit alpha B6259_RS01785 2657 2968 3175 2572 0.73 1.31 EtfB, electron transfer flavoprotein subunit beta B6259_RS01780 3996 4830 4357 4169 0.71 1.31 CAT, butyryl-CoA: acetate CoA-transferase B6259_RS06345 521 1497 283 330 1.55 4.01 c Fructose fermentation pathway PPF, 1-phosphofructokinase B6259_RS00100 276 2174 1256 239 0.35 7.33 Starch and sucrose metabolism PYG, glycogen phosphorylase B6259_RS00300 90 163 121 103 0.66 1.71 MalQ, 4-alpha-glucanotransferase B6259_RS07805 53 270 55 61 0.85 4.34 PGM, Phosphoglucomutase B6259_RS09200 95 189 127 113 0.66 1.80 Energy conservation energy-coupling factor transporter ATPase B6259_RS02790 141 104 117 159 1.04 0.76 electron transport complex protein RnfA B6259_RS06245 230 162 357 362 0.58 0.52 Sporulation stage 0 sporulation protein B6259_RS00205 379 279 233 252 0.97 0.82 stage II sporulation protein D B6259_RS09065 98 59 96 53 0.97 1.29 stage III sporulation protein AD B6259_RS03910 126 54 87 26 1.67 1.27 stage IV sporulation protein A B6259_RS04975 65 30 58 16 1.42 1.58 stage V sporulation protein AC B6259_RS09190 89 46 77 40 0.99 1.42 stage V sporulation protein AD B6259_RS09195 69 41 66 34 1.05 1.57 stage V sporulation protein AE B6259_RS00500 292 226 200 167 1.15 1.02 sporulation transcription factor Spo0A B6259_RS05505 127 115 83 106 0.94 0.94 sporulation transcriptional regulator SpoIIID B6259_RS01550 213 188 140 207 0.79 1.01 sporulation protein YtfJ B6259_RS04885 291 183 145 159 1.00 0.65 Transporter genes ABC transporter permease B6259_RS00445 446 274 27 235 5.17 c 1.27 metal ABC transporter B6259_RS00450 699 628 30 457 5.69 c 1.52 ABC transporter permease B6259_RS02670 296 130 441 387 0.60 0.40 b ABC transporter permease B6259_RS02665 180 96 258 231 0.62 0.48 b carbohydrate ABC transporter permease B6259_RS07005 33 124 90 41 0.37 b 3.51 c carbohydrate ABC transporter permease B6259_RS07905 71 744 71 40 0.90 12.71 c carbohydrate ABC transporter permease B6259_RS07810 39 229 40 45 0.85 5.48 c carbohydrate ABC transporter permease B6259_RS02030 26 71 16 39 1.35 2.14 c sugar ABC transporter permease B6259_RS07910 82 1175 88 50 0.86 14.74 c sugar ABC transporter permease B6259_RS03335 39 401 26 61 1.30 5.61 c sugar ABC transporter permease B6259_RS07815 36 197 37 49 0.85 4.34 c sugar ABC transporter permease B6259_RS07000 31 135 113 38 0.30 b 3.48 c iron ABC transporter permease B6259_RS00320 53 1278 77 89 0.62 10.05 c ABC transporter ATP-binding protein B6259_RS00440 504 277 51 239 5.25 c 1.39 ABC transporter ATP-binding protein B6259_RS00325 60 2032 94 100 0.58 11.14 c ABC transporter ATP-binding protein B6259_RS08900 153 682 233 214 0.58 3.13 ABC transporter ATP-binding protein B6259_RS07940 190 40 259 94 0.66 0.42 carbohydrate ABC transporter substrate-binding protein B6259_RS07915 216 3434 203 103 0.93 14.51 maltose ABC transporter substrate-binding protein B6259_RS03345 30 501 22 37 1.15 7.65 ABC transporter substrate-binding protein B6259_RS07820 372 1913 451 344 0.73 4.63 sugar ABC transporter substrate-binding protein B6259_RS02005 30 93 29 48 0.92 2.29 peptide ABC transporter substrate-binding protein B6259_RS08515 53 78 98 369 0.48 0.28 peptide ABC transporter substrate-binding protein B6259_RS02685 1385 819 1442 2222 0.85 0.50 ABC transporter ATP-binding protein B6259_RS02660 238 119 369 320 0.58 0.45 ABC transporter ATP-binding protein B6259_RS07940 190 58 259 166 0.66 0.42 PTS fructose transporter subunit IIC B6259_RS00095 372 2117 1273 485 0.37 3.87 PTS glucose transporter subunit IIA B6259_RS09280 484 260 1200 517 0.37 0.62 PTS β-glucoside transporter subunit IIABC B6259_RS01415 81 760 134 141 0.54 4.70 PTS mannitol transporter subunit IICBA B6259_RS00370 29 89 19 44 1.26 2.34 ferrous iron transport protein B B6259_RS03880 471 389 531 150 0.81 2.72 a, Data presented as mean of independent triplicates b, Significantly upregulated (FC ≥ 2.0, p < 0.05) c, Significantly downregulated (FC ≤ 0.5, p < 0.05) Expression of butyrate- and CA-producing genes The bioproduction of CA is a well-known chain elongation process from acetate (C2) to caproate (C6) via the reverse β-oxidation pathway, in which an acetyl-CoA (from ethanol) unit is combined with another acetyl-CoA (from acetate), and consequently C2 is elongated to C4, and further C4 is elongated to C6 [ 4 ]. Thus, acetyl-CoA is a key intermediate of flux distribution for the chain elongation. The conversion of pyruvate into acetyl-CoA is mainly catalyzed by the pyruvate: ferredoxin (flavodoxin) oxidoreductase (Pfor) that is a flavodoxin- and NADPH-dependent enzyme [ 6 , 26 ]. Here, the Pfor gene (B6259_RS09135) maintained at very high expression level (TPM > 4000) in the culture with lactate supplementation (Table 3 ), which were upregulated by 1.83- and 3.26-fold than that in the control in the growth and stationary phase, respectively (Fig. 7 ). High-level expression of the Pfor gene would result in increased acetyl-CoA, which provides the high amount of acetyl-CoA for chain elongation from acetate to butyrate and CA. Key enzymes involved in the butyrate formation include acetyl-CoA C-acetyltransferase (AtoB), 3-hydroxybutyryl-CoA dehydrogenase (HBD), enoyl-CoA hydratase (CRT), NAD-dependent butyryl-CoA dehydrogenase (BCD), Electron transfer flavoprotein (Etf) and butyryl-CoA: acetate CoA transferase (CAT) [ 6 , 17 ]. Here, genes encoding AtoB (B6259_RS06365), CRT (B6259_RS06360) and HBD (B6259_RS06355) were identified from the transcriptomes of strain CPB6, whose expression levels maintained at very high levels (TPM > 3000) in the culture with lactate supplementation throughout the growth and stationary phases, and were upregulated by 3.5–8.6 folds compared to the control. It suggested that the high-level expression of the three genes can be induced by supplemented lactate. In addition, two BCD genes (B6259_RS01790 and _RS02600) and EtfAB (B6259_RS01785 and _RS01780) responsible for the conversion of crotonyl-CoA to butyryl-CoA showed different expression profiles. B6259_RS01790 was expressed at a very high level (TPM > 3000) throughout the fermentation phases, but showing no change in expression in the two cultures. B6259_RS02600 was expressed at relatively low level at the growth phase, but its expression was induced to high levels in the culture with lactate supplementation at the stationary phase, which was > 4.4-fold higher than that in the control. More research is needed to determine which BCD gene plays the key role in acidogenesis in the CPB6. EtfAB showed no significant change in the two cultures during the growth and stationary phases. A CAT gene (B6259_RS06345) was detected in the CPB6 transcriptome, and its expression was markedly upregulated by 4-fold in the culture with lactate supplementation than that in the control in the stationary phase, and kept at very high expression levels. CAT is key enzyme responsible for catalysing the last step of the butyrate formation [ 26 ]. High-level expression of CAT gene should theoretically result in a high concentration of butyric acid in the culture with lactate supplementation. Nevertheless, significant accumulation of CA instead of butyric acid was observed in the CPB6 culture with lactate supplementation, suggesting that the CAT is likely more intent to transforming caproy-CoA to caproate instead of converting butyryl-CoA to butyrate. Up to now, little is known about the key functional genes responsible for CA synthesis from butyryl-CoA. Genes involved in butyrate synthesis via the reverse β-oxidation (e.g., AtoB, CRT, HBD, BCD and CAT) are assumed to have the function in the caproyl-CoA and CA formation [ 6 ]. However, Clostridium sp. BPY5 and C . tyrobutyricum , which contains these genes, only produce butyric acid instead of CA [ 26 , 27 ], while C . kluyveri and Ruminococcaceae bacterium CPB6, which contain these genes, can further elongate butyric acid to CA [ 6 , 16 ]. It suggested that there may be differences in structure and function between these genes from different organisms. Therefore, the further study needs to be performed to explore the functions of these genes in strain CPB6. Lactate is a major endproduct of glycolysis in the absence of oxygen [ 28 ]. Its formation or conversion requires lactate dehydrogenase (LDH) with the regeneration of NADH to NAD+. There are two LDH genes in the CPB6 genome [ 17 ], encoding L-lactate dehydrogenase (L-LDH) and D-lactate dehydrogenase (D-LDH), respectively. In this study, the expression of LDH genes (L-LDH, B6259_RS09845; D-LDH, B6259_RS06770) were also detected in the transcripts of strain CPB6. However, the two genes showed relatively low expression levels in both the culture with lactate supplementation and the control, except that slightly higher expression level was observed in the control at the stationary phase (Table 3 ). LDH catalyzes the reaction converts pyruvate to lactate or the reverse reaction that converts lactate to pyruvate [ 29 ]. This conversion is essential in hypoxic and anaerobic conditions when ATP production by oxidative phosphorylation is disrupted. The recent study showed that lactate can be transformed into CA in either mixed microbiome [ 2 , 13 , 14 ], or in the pure culture where the conversion of lactate to acetyl-CoA coupling with the reverse β-oxidation is speculated to result into chain elongation [ 15 ]. Interestingly, the lactate supplementation in this study did not lead to increased expression levels of LDHs in either growth or stationary phases, indicating that the expression of LDHs might be uncoupled from the utilization of lactate. It warrants further investigation concerning the function of LDH in the conversion of lactate to CA in the CPB6 strain. The gene encoding phosphate acetyltransferase (PTA, B6259_RS07830), one important enzyme involved in acetate formation, was remarkably upregulated in the culture with lactate supplementation than in the control. However, the expression of acetate kinase (ACK, B6259_RS03430) showed no change in response to the addition of lactate. By including the production of H 2 and CO 2 into the loop, it could provide a whole picture for carbon balance for the substrate utilization and cell biomass production. Unfortunately, the production of H 2 and CO 2 was not monitored in this study. In the future studies, this should be taken into consideration for improvement. Expression of putative ABC transporter and sporulation genes Strain CPB6, affiliated with Clostridium cluster IV of the family Ruminococcaceae in the order of Clostridiales, is a spore-forming, obligate anaerobic bacterium that can produce CA from lactate (Zhu et al. 2017). As shown in Table 3 , sporulation genes showed similar expression patterns in both groups, e.g., spo 0, spo IIID, spo V, spo YtfJ, were induced to high expression under both conditions (with or without lactate supplementation) at the growth and stationary phases, while spo IID, spo IIIAD, spo IVA, spo VAC, spo VAD spo VAE were expressed at low or moderate levels. Some bacteria, such as bacilli and clostridia, develop into highly resistant spores to protect their genome and cell from certain doom when living conditions become intolerable [ 30 ]. It ensures bacterial survival under adverse environmental conditions. Sporulation in Clostridium spp. is ordinarily not triggered by starvation but by cessation of growth in the presence of excess carbon source or exposure to oxygen [ 31 ]. The two most critical factors involved in the shift to solventogenesis, a decrease in external pH and accumulation of acidic fermentation products, are generally assumed to be associated with the initiation of sporulation in Clostridium spp., to some extent [ 32 ]. But recent studies showed that the sporulation events were uncoupled from the induction of solventogenesis in C. beijerinckii [ 23 ]. In this study, the sporulation genes showed no significant difference between the culture with lactate supplementation and the control, indicating that the sporulation events are not associated with the production of CA in the CPB6 until the stationary phase. This may be because low concentrations of CA (1717 mg/L) are not sufficient to initiate sporulation for the CPB6. It will be investigated in the future whether high concentrations of CA trigger sporulation. In the transcriptome of strain CPB6, most genes encoding ATP-binding cassette (ABC) transporters and substrate-bind proteins (SBP) maintained at low expression levels in the control, but were induced to relatively high expression levels (particularly upregulated by 2–14 folds at the stationary phase) in the culture with lactate supplementation. ABC transporters are ubiquitous membrane proteins that couple the transport of diverse substrates across cellular membranes to the hydrolysis of ATP [ 33 ]. ABC transporters are generally divided into importers and exporters on the basis of the polarity of solute movement. ABC importers are found mostly in bacteria and are crucial in mediating the uptake of solutes including sugar, metal ions and vitamins [ 34 ]. In the present study, most genes for ABC transporter and substrate-binding protein (SBP) were no significant change in the two cultures, except two ABC transporter genes (B6259_RS00445, B6259_RS00450), and one SBP gene (B6259_RS00440) which were upregulated by more than 2-fold in the culture with lactate supplementation than in the control. It was suggested that the three genes might be related to the intake and use of lactic acid. In addition, nine ABC transporter genes and six SBP genes were markedly upregulated at the stationary phase. Specially, B6259_RS07905, _RS07910, _RS00320, _RS00325 and B6259_RS07915 were increased over 10-fold in the culture with lactate supplementation than in the control, demonstrating that these genes are associated with the extrusion of CA from the cell and the maintenance of osmotic homeostasis in cytoplasm [ 35 ]. In addition, two phosphotransferase system (PTS) transporter genes (B6259_RS01415 and B6259_RS00370) and one ferrous iron transporter gene (B6259_RS03880) were upregulated by 2- to 4-fold in the culture with lactate supplementation than in the control. PTS is a multiple-component carbohydrate uptake system that drives specific saccharides across the bacterial inner membrane while simultaneously catalyzing sugar phosphorylation [ 36 ]. Five distinct subfamilies of proteins related to PTS have been identified within the glucose superfamily: the lactose family, the glucose family, the β-glucoside family, the mannitol family, and the fructose family [ 37 ]. In this study, four PTS transporter genes were detected in the transcriptome of strain CPB6, including PTS fructose transporter subunit IIC (B6259_RS00095), PTS glucose transporter subunit IIA (B6259_RS09280), PTS β-glucoside transporter subunit IIABC (B6259_RS01415), and PTS mannitol transporter subunit IICBA (B6259_RS00370). Genes encoding PTS fructose and glucose transporters were highly expressed under both conditions, but the two genes were significantly downregulated at the growth phase in the culture with lactate supplementation than in the control, indicating that PTS transporter-mediated sugar transport in membrane vesicles in CPB6 is inhibited by the lactate supplementation. However, at the stationary phase, genes encoding PTS fructose, β-glucoside and mannitol transporters were all strikingly upregulated. Overall, the addition of lactate caused distinct changes in gene expression of the CPB6 strain, especially at stationary phase. The detailed mechanism remains to be further studied. Conclussions This study showed that lactate supplementation induced earlier CA production, higher CA titer and productivity, but had little impact on cell growth, suggesting that the supplemented lactate is more involved in secondary metabolism (e.g., the production of CA) than the carbon assimilation. The gene transcriptional profiles based on RNA-Seq demonstrated that supplemented lactate promoted CA production by altering the expression patterns of genes involved in crucial metabolic pathways. Specifically, the Pfor gene involved in central pyruvate metabolism, 5 genes (AtoB, HBD, CRT, BCD and CAT) involved in the reverse β-oxidation pathway, 11 genes encoding ABC transporter, 6 SBP genes, and 4 PTS transporter genes showed sharp upregulation in response to the addition of lactate. The findings presented herein provide unique insights into the metabolic effects of lactate on CA production at the gene regulation level. Methods Microorganisms, media and fermentation experiment Ruminococcaceae bacterium CPB6 (GDMCC No.60133) was routinely cultured at 37 o C anaerobically in a modified tryptone-glucose-yeast extract (mTGY) medium containing the following compounds (g L − 1 , pH 6.0): tryptone, 5.0; glucose, 2.0; yeast extract, 5.0; sodium acetate, 3.5; K 2 HPO 4 ·3H 2 O, 0.41; KH 2 PO 4 , 0.23; NH 4 Cl, 0.25; MgSO 4 ·7H 2 O, 0.20; NaHCO 3 , 2.5; L-cysteine, 0.25; Na 2 S·9H 2 O, 0.25; resazurin, 0.0005; and 1 ml of trace element solution SL-10 and 1 ml of vitamin solution [ 14 ]. The suspension of activated strain CPB6 was inoculated with a 5% ratio into the same medium as described above and incubated for 12–15 h until the OD 600 of the culture reached 0.8-1.0. Then the culture would be used as the seed inoculum (5% ratio, v/v) for batch experiments. To investigate the effect of lactate on cell growth and CA production, 5 g L − 1 sodium lactate was supplemented into the modified TGY medium (mTGYL). Batch experiments were performed in 250 mL serum bottles containing 100 mL of modified mTGY or mTGYL media. The headspace of the bottle was filled with highly pure N 2 . Each fermentation was performed in triplicate. The fermentation was carried out at 37 o C for 24 h in an E500 anaerobic workstation (Gene Science, USA) under N 2 : CO 2 : H 2 (volume ratio of 80:10:10) atmosphere. Samples were taken at specific times and processed for cell concentration determination and HPLC analysis. Samples for RNA isolation were taken at the growth and stationary phases of the cell growth. Analytical methods Culture growth was monitored by measuring the optical density at 600 nm (OD 600 ) using a TU-1810 UV/Vis Spectrophotometer (Puxi Instrument Co. Ltd., Beijing). Lactic acid, acetic acid, butyric acid, and caproic acid were quantified using an HPLC system (Agilent 1260 Infinity, USA) equipped with a differential refraction detector (RID) and a Agilent Hi-Plex H column (300 × 6.5 mm) following the procedure as previously described [ 16 ]. RNA isolation, library construction, and sequencing In preparation for RNA isolation, 10 ml cell culture was harvested at each time point, and centrifuged at 8,000 × g for 10 min at 4 °C. Cells were then frozen in liquid nitrogen prior to storage at -80 °C. The RNA was extracted and purified using a RNA extraction kit (DP430, Tiangen Biotech, Beijing, China) following the manufacturer's protocol. RNA quality and quantity were characterized using a NanoDrop2000 (NanoDrop Technologies, Wilmington, DE), agarose gel electrophoresis (RNA integrity detection) and Agilent 2100 (RIN value measurement). Only the RNA samples with high-quality (≥ 5 µg; ≥200 ng/µL; OD260/280 = 1.8 ~ 2.2) were used for the cDNA library construction and sequencing. Before library construction, rRNAs were removed with the Ribo-Zero rRNA Removal Kit (Epicentre, San Diego, CA) following the manufacturer’s protocol. The enriched mRNA was fragmented into short fragments (approximately 200 bp) randomly using metal ions. Then the first strand cDNA was synthesized using the random hexamer-primer with the mRNA fragment as the template. After synthesizing the second strand cDNA using DNA polymerase I and RNase H, double-stranded cDNA was further end repaired, A-tailed, and indexed adapters ligated. The final cDNA library was constructed using the mRNA-Seq library construction kit (Illumina Inc., San Diego, CA), and then sequenced by Illumina platform (Illumina Inc., San Diego, CA) with 2 × 150 bp. RNA-Seq data analysis High quality reads in each sample were aligned to the CPB6 genome using Bowtie2, and those that did not align uniquely to the genome were discarded using the default quality parameters. Each base was assigned a value based on the number of mapped sequence coverage. Gene expression levels were defined using the number of transcripts per million (TPM), which is proportional to the quantity of cDNA fragments derived from the gene transcripts. The quantitative gene expression values between samples were identified by calculating the number of unambiguous tags for each gene and then normalizing this to TPM, which was calculated following the method reported by Parto et al., [ 38 ]. The gene expression results were visualized using heatmap plots. Statistical analysis Significant differences of the gene expression between the culture from fermentation with lactate supplementation vs. the control were determined using ANOVA in R software (version 3.5.2). TPM values were first transformed to log10-scale. The log10-transformed TPM values were then properly centered for better representation of the data using the heatmap plots. Fold changes (FCs) as the ratio of the TPM values were calculated following the method reported by Love et al. [ 39 ], and were used to compare the differentially expressed genes (DEGs) between the culture from fermentation with lactate supplementation and the control. Declarations Availability of data and materials The RNA-Seq sequencing data have been deposited in the NCBI Sequence Read Archive (SRA) under the accession number PRJNA564589 Ethics approval and consent to participate Not applicable. Competing interests The authors declare that they have no competing interests. Funding This work has been supported by the Natural Science Foundation of China (31770090), the Open-foundation project of CAS Key Laboratory of Environmental and Applied Microbiology (KLCAS-2017-01). 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Supplementary Files TableS2.xlsx TableS1.xlsx FigS2.jpg FigS1.jpg Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Major revision 27 Jan, 2021 Review # 2 received at journal 04 Jan, 2021 Review # 3 received at journal 04 Jan, 2021 Reviewer # 3 agreed at journal 13 Dec, 2020 Reviewer # 2 agreed at journal 13 Dec, 2020 Review # 1 received at journal 05 Dec, 2020 Reviewers invited by journal 23 Oct, 2020 Reviewer # 1 agreed at journal 23 Oct, 2020 Editor assigned by journal 15 Sep, 2020 Submission checks completed at journal 14 Sep, 2020 Editor invited by journal 14 Sep, 2020 First submitted to journal 10 Sep, 2020 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-76088","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research article","associatedPublications":[],"authors":[{"id":2529290,"identity":"af91b240-a094-47d6-8f44-922f9447aab4","order_by":0,"name":"Yong Tao","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAwUlEQVRIiWNgGAWjYBADOQZmHiKVwtQZk64lsYGBWC327IePSXzcUZve3857gOFHDYO8OUFbeNLSJGeeOZ474zBfAmPPMQbDnQ0EHZZjJs3bdix3AzOPAQNvA0OCwQFCWvjfgLWkGwC1MP4lSosE2JaaBJAWZuJsufEs2XJm2wFDkF8OyxyTMNxASAt7f/LBGx/b6uT5+88efPimxkaeoC1AwCLBwHAYzAIqliCsHgiYPzAw1BGlchSMglEwCkYoAABJuTksCIDGnAAAAABJRU5ErkJggg==","orcid":"","institution":"CIB CAS: Chengdu Institute of Biology","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Yong","middleName":"","lastName":"Tao","suffix":""},{"id":2529291,"identity":"a6b7bb4e-2146-496d-bb5c-90b9d74f33b3","order_by":1,"name":"Shaowen Lu","email":"","orcid":"","institution":"Chengdu Institute of Biology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Shaowen","middleName":"","lastName":"Lu","suffix":""},{"id":2529292,"identity":"e26ea589-0342-4c3a-9d96-932fb83a39b9","order_by":2,"name":"Yi Wang","email":"","orcid":"","institution":"Auburn University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yi","middleName":"","lastName":"Wang","suffix":""},{"id":2529293,"identity":"cccebe0e-5bda-4fbf-b0d7-882dfbe5864c","order_by":3,"name":"Cuicui Wei","email":"","orcid":"","institution":"Chengdu Institute of Biology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Cuicui","middleName":"","lastName":"Wei","suffix":""},{"id":2529294,"identity":"d2144286-9f75-42ac-b636-0b36d3574fb0","order_by":4,"name":"Hong Jin","email":"","orcid":"","institution":"Chengdu Medical College","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hong","middleName":"","lastName":"Jin","suffix":""},{"id":2529295,"identity":"7b27e42d-b7cd-4b2a-a978-9af0035a95e9","order_by":5,"name":"Xiaohong He","email":"","orcid":"","institution":"Chengdu Institute of Biology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Xiaohong","middleName":"","lastName":"He","suffix":""}],"badges":[],"createdAt":"2020-09-11 10:24:32","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-76088/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-76088/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":2537751,"identity":"6b265e42-2272-4b86-a203-fef742b29e88","added_by":"auto","created_at":"2020-09-22 15:45:31","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":484622,"visible":true,"origin":"","legend":"Fermentation kinetics of Ruminococcaceae bacterium CPB6. (a) Cell growth profiles. Time points for taking samples subjected to RNA-Seq are indicated with red vertical arrows. L: fermentation with lactate supplementation; C: control fermentation without lactate supplementation; (b) Sugar consumption and metabolites production during the fermentation with the supplementation of lactate; (c) Sugar consumption and metabolites production during the control fermentation (without the supplementation of lactate). Values represent the mean of the biological triplicates and error bars represent the standard deviations.","description":"","filename":"Figure1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-76088/v1/Figure1.jpg"},{"id":2537752,"identity":"c075bb06-f892-46eb-9cf5-16c35e4160f0","added_by":"auto","created_at":"2020-09-22 15:45:32","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":223046,"visible":true,"origin":"","legend":"Frequency histogram of transcripts from the RNA-Seq results. L: cell culture with lactate supplementation; C: control culture without lactate supplementation; Number 1 and 2 represented the growth phase and stationary phase, respectively. The diagram shows the distribution of the number of genes expressed at different transcripts per million (TPM) levels. The percentage value above each bar indicates the genes at the specific expression level accounting for the proportion of the total number of genes. The ‘∗’ mark indicates that significantly different frequencies (i.e., numbers of genes) were observed between the two RNA-Seq data sets from lactate-supplemented fermentation (L) vs. the control (C), respectively. ","description":"","filename":"Figure2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-76088/v1/Figure2.jpg"},{"id":2537753,"identity":"83817fdc-3fd6-48ad-8a36-27d946fe41b3","added_by":"auto","created_at":"2020-09-22 15:45:32","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":284465,"visible":true,"origin":"","legend":"Annotation of genes using Gene Ontology (GO) in the transcriptome of strain CPB6. Left axis: the proportion of genes falling into each GO category; right axis: the number of genes falling into each GO category.","description":"","filename":"Figure3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-76088/v1/Figure3.jpg"},{"id":2537754,"identity":"879e3a5a-1e42-4f54-ba9c-6c0fde84da82","added_by":"auto","created_at":"2020-09-22 15:45:32","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":486060,"visible":true,"origin":"","legend":"The number of differentially expressed genes (DEGs) between the culture with lactate supplementation and the control falling into each Clusters of Orthologous Groups (COG) categories during the growth phase (a) and the stationary phase (b), respectively. Note that since COG categories overlap, the sum of all COG annotated genes are larger than the total number of all up- and down-regulated genes analyzed.","description":"","filename":"Figure4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-76088/v1/Figure4.jpg"},{"id":2537755,"identity":"33a5248a-a257-41ae-bc5a-031d44ba1cd4","added_by":"auto","created_at":"2020-09-22 15:45:32","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":308977,"visible":true,"origin":"","legend":"Clustering of differentially expressed genes (DEGs) between the culture with lactate supplementation (L) and the control (C) at the growth and stationary phases. Values of log10TPM were conducted at normalized transformation before clustering. Red indicates genes with high expression, and blue indicates genes with low expression. Each column represented a sample, and each row represented a gene. The left was the tree diagram of clustering gene, and the upper part was the tree diagram of sample clustering, and the bottom was the name of each sample (L: the culture with lactate supplementation, C: the control; Number 1 and 2 represented growth phase and stationary phase, respectively; a, b and c represented the biological triplicate samples).","description":"","filename":"Figure5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-76088/v1/Figure5.jpg"},{"id":2537756,"identity":"2947b107-b125-4674-8e0f-9c124d6fc9bf","added_by":"auto","created_at":"2020-09-22 15:45:32","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":371227,"visible":true,"origin":"","legend":"Venn diagram of the numbers of differentially expressed genes (DEGs) trigger by substrate type (plus/minus lactate) vs growth stage (stationary phase vs growth phase) (a), the numbers of DEGs trigger by substrate type vs growth stage (b). The overlap of circles was defined as genes affected by both substrate type and growth stage.","description":"","filename":"Figure6.jpg","url":"https://assets-eu.researchsquare.com/files/rs-76088/v1/Figure6.jpg"},{"id":2537757,"identity":"4a979e37-3a39-49db-96ae-3f96b90f00d8","added_by":"auto","created_at":"2020-09-22 15:45:32","extension":"jpg","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":4841767,"visible":true,"origin":"","legend":"The overview of the central metabolic pathway in strain CPB6 with fold changes (FCs) of the expression of genes based on the RNA-Seq results. The number in parentheses represents the FC of TPM between the culture with lactate supplementation vs. the control; the first and the second numbers in the same parentheses represent the gene expression FC in the growth and stationary phases, respectively. Black solid lines symbolize enzymatic reactions. Red dashed lines mark enzymatic reactions for which corresponding enzymes are probably not encoded in strain CPB6.","description":"","filename":"Figure7.jpg","url":"https://assets-eu.researchsquare.com/files/rs-76088/v1/Figure7.jpg"},{"id":15669045,"identity":"d599e1a5-bb08-4dbb-a647-7493fb2503ed","added_by":"auto","created_at":"2021-11-18 13:51:25","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1633827,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-76088/v1/643bb22c-0d0b-4ba1-b0a3-7cc056903049.pdf"},{"id":2537759,"identity":"1cc111a0-771c-4194-a856-04b6d5be4d36","added_by":"auto","created_at":"2020-09-22 15:45:33","extension":"xlsx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":31717,"visible":true,"origin":"","legend":"","description":"","filename":"TableS2.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-76088/v1/TableS2.xlsx"},{"id":2537760,"identity":"16040b6b-d39c-40a4-b3f2-917a695937f1","added_by":"auto","created_at":"2020-09-22 15:45:33","extension":"xlsx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":10669,"visible":true,"origin":"","legend":"","description":"","filename":"TableS1.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-76088/v1/TableS1.xlsx"},{"id":2537761,"identity":"31e14390-1ff1-463b-849d-a76b78dc6034","added_by":"auto","created_at":"2020-09-22 15:45:33","extension":"jpg","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":5267335,"visible":true,"origin":"","legend":"","description":"","filename":"FigS2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-76088/v1/FigS2.jpg"},{"id":2537762,"identity":"903fc4d3-ce6a-4099-9057-bc7b841045e3","added_by":"auto","created_at":"2020-09-22 15:45:34","extension":"jpg","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":9585027,"visible":true,"origin":"","legend":"","description":"","filename":"FigS1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-76088/v1/FigS1.jpg"}],"financialInterests":"","formattedTitle":"\u003cp\u003eGenome-wide transcriptomic analysis of \u003cem\u003en\u003c/em\u003e-caproic acid production in \u003cem\u003eRuminococcaceae\u003c/em\u003e bacterium CPB6 with lactate supplementation\u003c/p\u003e","fulltext":[{"header":"Background","content":" \u003cp\u003eThe increasing demand for fuels and chemicals and the scarcity of fossil resources necessitate the development of sustainable and innovative strategies for the industrial production. Organic residual streams (e.g., food waste and brewery wastewater) have great potential to be employed as feedstock for the production of biofuels and biochemicals [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cem\u003en\u003c/em\u003e-Caproic acid (CA) is a medium chain fatty acid of 6-carbon, which has recently gained considerable attention due to its high value as a fuel and chemical feedstock [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Biosynthesis of CA can be achieved in some anaerobic bacteria (e.g., \u003cem\u003eClostridium kluyveri\u003c/em\u003e) via the chain elongation pathway with ethanol as reduced substrate (electron donor) [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e], in which the oxidation of ethanol provides energy and acetyl-CoA for the reverse β-oxidation pathway [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Many studies show that the addition of ethanol during the acidification of wastes can promote chain elongation, and lead to higher volumetric production rates and a high CA selectivity [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e], indicating that ethanol is an efficient reduced substrate for CA production. In addition, researchers have explored other chemicals used as electron donors for CA production, including hydrogen [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e], methanol [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e], propanol [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e], and \u003cem\u003eD\u003c/em\u003e-galactitol [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eRecently, lactate was showed to be a potential alternative to ethanol for the production of CA by reactor microbiome systems [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. More recently, our study demonstrates that CA is efficiently produced from lactate by the pure culture of a \u003cem\u003eRuminococcaceae\u003c/em\u003e bacterium CPB6 isolated from a fed-batch reactor microbiome [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Strain CPB6 is an anaerobic mesophilic bacterium that is highly prolific in its ability to perform chain elongation of lactate to caproate, and can produce CA (C6) from lactate (as electron donor) with C2-C4 carboxylic acids as electron acceptors, and heptoic acid (C7) with C3-C5 carboxylic acids [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. The phylogenic analysis based on 16S rRNA sequences and the whole genome showed that strain CPB6 might belong to a new clade (genus) of the family Ruminococcaceae, it is thus tentatively christened with the name Ruminococcaceae bacterium CPB6 [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Furthermore, we identified sets of genes correlated with the chain elongation pathway by sequencing and annotating the whole genome of the CPB6 strain. However, to date, very little information is available on genome-wide transcriptomic analysis of the CPB6 strain for CA production using lactate as the electron donor, which is essential to understand the effect of lactate on the metabolic pathway shift for CA production at the molecular level and thus elucidate proper strategies for further strain improvement.\u003c/p\u003e \u003cp\u003eRNA-Seq is a powerful method to elucidate the metabolism and identify specific genes/enzymes for the particular interesting metabolic pathways [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. In the present study, RNA-Seq for the transcriptomic analysis of strain CPB6 was carried out to investigate the effect of lactate supplementation on cell growth, CA production and other essential metabolisms. The identified key genes herein provide candidates for the metabolic engineering in the future in order to develop more robust strains based on CPB6 for enhanced production of CA and relevant biochemicals.\u003c/p\u003e "},{"header":"Results And Discussion","content":" \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eGrowth and the production of CA\u003c/h2\u003e \u003cp\u003eAs shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ea, cells took approximately 18\u0026nbsp;h to grow to the stationary phase. Although the maximum OD\u003csub\u003e600\u003c/sub\u003e of the culture with lactate supplementation was slightly higher than that of the control (without lactate supplementation) at the stationary phase (1.25 vs 1.16), both cultures showed similar growth kinetics, indicating that lactate supplementation had little impact on cell growth. The CA production was started to be observed in the culture with lactate supplementation after 6\u0026nbsp;h of cultivation, and the CA titer continued to increase and reached 1717.2\u0026nbsp;mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e at 21\u0026nbsp;h (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eb), while CA production was not detected until 15\u0026nbsp;h of cultivation in the control, the CA titer of which only reached 618\u0026nbsp;mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e at 21\u0026nbsp;h. In both cultures, while more significantly in the culture with lactate supplementation, the main endproduct was CA together with small amount of butyrate, suggesting that the carbon flux of acetyl-CoA may be inclined to be channelled to CA synthesis, especially in the presence of lactate. This is consistent with our previous studies [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eIn sum, compared to the control without lactate supplementation, the lactate supplementation had little effect on the cell growth, but led to earlier initiation for CA production (6 vs 15\u0026nbsp;h), higher final CA titer (1717 vs 618\u0026nbsp;mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) and higher CA productivity (81.8 vs 29.4\u0026nbsp;mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e h\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eRNA-Seq statistics\u003c/h2\u003e \u003cp\u003eSamples were taken for RNA-Seq analysis from both the growth (12\u0026nbsp;h) and stationary (18\u0026nbsp;h) phases for both the culture with lactate supplementation and the control. For each culture, independent biological triplicates (a, b, and c) were included (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Therefore, a total of twelve samples were taken for cDNA libraries construction and sequencing on the Illumina HiSeq platform (Illumina, San Diego, USA). The number of raw reads generated from the sequencing for each library was from 15.7 to 23.5\u0026nbsp;million (Table S1 in Additional files). A total of 224\u0026nbsp;Mb sequence reads from 12 cDNA libraries were mapped to the genome of strain CPB6. Only those reads that mapped unambiguously to the CPB6 genome were used for further analysis.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eSummary of RNA-Seq sequencing and data analysis results\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"13\"\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eSample Name\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c4\" namest=\"c2\"\u003e \u003cp\u003eL1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c7\" namest=\"c5\"\u003e \u003cp\u003eL2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c10\" namest=\"c8\"\u003e \u003cp\u003eC1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c13\" namest=\"c11\"\u003e \u003cp\u003eC2\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003ea\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eb\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003ec\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003ea\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003eb\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003ec\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003ea\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003eb\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003ec\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u003cb\u003ea\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u003cb\u003eb\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cb\u003ec\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTotal reads\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e22982792\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e18614536\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e18087962\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e16881648\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e16844058\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e15343814\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e18684112\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e19108148\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e19099082\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e19909258\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e23032120\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e18662552\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo. of read mapped\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e22684858\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e18390709\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e17865180\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e16713319\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e16606726\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e15098988\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e18437261\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e18893873\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e18905208\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e19631686\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e22572873\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e18285805\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRatio of reads mapped (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e98.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e98.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e98.77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e98.59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e98.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e98.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e98.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e98.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e98.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e98.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e97.98\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo. of unique reads mapped\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e22339068\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17915954\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e17559600\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e16540060\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e16324768\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e14862898\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e18050316\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e18505697\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e18508661\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e19247256\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e22187272\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e17856892\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo. of genes with detectable expression\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1969\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1968\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1968\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1968\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1969\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1968\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1968\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e1968\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e1968\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e1968\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e1969\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e1969\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRange in expression levels (TPM)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.3\u0026ndash;2.7\u0026thinsp;\u0026times;\u0026thinsp;10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8.0-1.7\u0026thinsp;\u0026times;\u0026thinsp;10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e11.5\u0026ndash;1.7\u0026thinsp;\u0026times;\u0026thinsp;10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.2\u0026ndash;5.3\u0026thinsp;\u0026times;\u0026thinsp;10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e26.4\u0026ndash;3.7\u0026thinsp;\u0026times;\u0026thinsp;10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6.6\u0026ndash;7.3\u0026thinsp;\u0026times;\u0026thinsp;10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e4.3-2.0\u0026thinsp;\u0026times;\u0026thinsp;10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3.8\u0026ndash;1.8\u0026thinsp;\u0026times;\u0026thinsp;10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e4.1\u0026ndash;1.9\u0026thinsp;\u0026times;\u0026thinsp;10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e5.2\u0026ndash;2.4\u0026thinsp;\u0026times;\u0026thinsp;10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e10.8\u0026ndash;1.8\u0026thinsp;\u0026times;\u0026thinsp;10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e24.5\u0026ndash;1.9\u0026thinsp;\u0026times;\u0026thinsp;10\u003csup\u003e4\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"13\"\u003eL1: cell culture with lactate supplementation from the growth phase;\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"13\"\u003eL2: cell culture with lactate supplementation from the stationary phase;\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"13\"\u003eC1: Control culture without lactate supplementation from the growth phase;\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"13\"\u003eC2: Control culture without lactate supplementation from the stationary phase.\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"13\"\u003ea, b and c represented the biological triplicate samples.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eOverall, out of reads derived from all the samples, 15.1 to 22.7\u0026nbsp;million reads were unambiguously mapped to the CPB6 genome, and over 98% reads were mapped (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). A total of 1968/1969 out of 2045 protein-coding genes had detectable expression in both conditions (with or without lactate supplementation) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e), indicating that the RNA-Seq analysis in this study achieved comprehensive coverage of the CPB6 transcriptome. The transcription levels (the number of transcripts per million, TPM) of most active protein-coding genes were in the range of 3.2\u0026thinsp;\u0026times;\u0026thinsp;10\u003csup\u003e4\u003c/sup\u003e\u0026ndash;7.3\u0026thinsp;\u0026times;\u0026thinsp;10\u003csup\u003e4\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eAs illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, the gene expression could be classified into four levels: low (TPM\u0026thinsp;\u0026lt;\u0026thinsp;30), moderate (TPM: 30\u0026ndash;150), high (TPM: 150\u0026ndash;1000), and very high (TPM\u0026thinsp;\u0026gt;\u0026thinsp;1000). The number of genes at some specific expression levels was significantly different for the two cultures (with or without lactate supplementation). For the growth phase, there were slightly more genes in the moderate and high and very high expression level in the culture with lactate supplementation than in the control, but lowly expressed genes were significantly decreased. While for the stationary phase, the culture with lactate supplementation had more genes in the moderate expression level, but fewer genes in the high and very high expression level. These results suggested that more changes in gene expression are triggered by the addition of lactate at the stationary phases.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eFunctional annotation and classification\u003c/h2\u003e \u003cp\u003eIn the transcriptome of strain CPB6, a total of 1122 expressed genes were allocated into three primary Gene Ontology (GO) categories (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e), including the category of biological process (601 genes), cellular component (524 genes), and molecular function (916 genes). In each category, the genes were further assigned into 28 functional groups, such as metabolic process (478 genes), cellular process (440 genes), cell part (307 genes), membrane part (297 genes), catalytic activity (654 genes), binding (561genes), and etc. The analysis of the genes based on the KEGG annotation identified a total of 1046 unigenes allocated into six primary KEGG categories including 35 subcategories (Figure. S1 in Additional files). The top 5 categories of genes were: carbohydrate metabolism, amino acid metabolism, membrane transport, translation, and metabolism of cofactors and vitamins, respectively. The top 10 enriched pathways included ABC transporters (58 genes), Amino sugar and nucleotide sugar metabolism (27 genes), Starch and sucrose metabolism (26 genes), Glycolysis / Gluconeogenesis (24 genes), Purine metabolism (40 genes), Pyrimidine metabolism (37 genes), Peptidoglycan biosynthesis, Aminoacyl-tRNA biosynthesis (28 genes), Ribosome (52 genes), and Quorum sensing (26 genes). The analysis based on the Clusters of Orthologous Groups (COGs) showed that 1785 unigenes were allocated to four primary COG categories containing 20 COG functional clusters. The top 5 annotated genes corresponding to the KEGG pathways were Replication recombination and repair (155 genes), Translation, ribosomal structure and biogenesis (142 genes), Amino acid transport and metabolism (126 genes), Carbohydrate transport and metabolism (110 genes), and Inorganic ion transport and metabolism (Figure S2 in Additional files).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eDifferential expression of global genes\u003c/h2\u003e \u003cp\u003eThe correct identification of differentially expressed genes (DEGs) between specific conditions is a key in the understanding phenotypic variation of organisms under environmental stress. As shown in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, there were only 34 differentially expressed genes (DEGs, FC\u0026thinsp;\u0026ge;\u0026thinsp;2 or \u0026le;\u0026thinsp;0.5 with P-value\u0026thinsp;\u0026lt;\u0026thinsp;0.05) identified during the growth phase between two culture conditions, of which 15 genes were upregulated, and 19 genes were downregulated more than two-fold. In addition, a total of 245 DEGs were identified at the stationary phase, of which 123 genes were significantly upregulated and 122 genes were downregulated (Table S2 in Additional files). It suggested that the addition of lactate led to differences in gene expression between the two cultures (with and without lactate supplementation) during different growth phases.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe differentially expressed genes in culture with/without lactate supplementation during the growth phase\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eNo.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eGene_ID\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eGene name\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eGene description\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003eTPM\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eFC (L1/C1)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eP-value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003eC1\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003eL1\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"8\" nameend=\"c8\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003e15 upregulated genes (FC\u0026thinsp;\u0026ge;\u0026thinsp;2.0); all statistically significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS06365\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eatoB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eacetyl-CoA C-acetyltransferase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1224\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5204\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e7.9E-39\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS06360\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ecrt\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eenoyl-CoA hydratase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e795\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3434\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2.4E-33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS06355\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ehbd\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3-hydroxybutyryl-CoA dehydrogenase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1418\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6306\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2.3E-27\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07830\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003epta\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ephosphate acetyltransferase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e271\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e666\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e4.3E-24\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00440\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003emethionine ABC transporter ATP-binding protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e504\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e6.2E-18\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00450\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003emetal ABC transporter substrate-binding protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e699\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5.69\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e5.9E-15\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00445\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eABC transporter permease\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e446\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e6.9E-14\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS08190\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ecysK\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ecysteine synthase A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e390\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e7426\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e4.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e6.4E-10\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS08440\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eunknown function\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1048\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3598\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e5.2E-06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS06010\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ehypothetical protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e8.1E-06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07140\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ehypothetical protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e154\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e470\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.0E-05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS01720\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ecadA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ecadmium-translocating P-type ATPase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.9E-05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS06870\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eHsp20/alpha crystallin family protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e315\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1102\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.6E-04\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00455\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003epepT\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003epeptidase T\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e576\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2.0E-04\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS02585\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ebdh\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ebutanol dehydrogenase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e242\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.1E-04\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"8\" nameend=\"c8\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003e19 downregulated genes (FC\u0026thinsp;\u0026le;\u0026thinsp;0.5); all statistically significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS08515\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003epeptide ABC transporter substrate-binding protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.7E-23\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09280\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePTS glucose transporter subunit IIA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e484\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e5.5E-23\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09735\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eilvH\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eacetolactate synthase small subunit\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e564\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e302\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e9.4E-19\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS06995\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ehypothetical protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e276\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2.7E-18\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS08565\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ehypothetical protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e143\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e2.3E-13\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003esugar ABC transporter permease\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e113\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e9.2E-13\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS01525\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eunknown function\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2683\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1010\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e8.4E-12\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS03200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eunknown function\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2683\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1010\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e8.4E-12\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07010\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003etag\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eglycosylase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e144\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e9.9E-11\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ecarbohydrate ABC transporter permease\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.1E-09\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS01865\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eDUF2520 domain-containing protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e260\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e6.7E-09\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS01880\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003epanD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003easpartate 1-decarboxylase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e444\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e156\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e9.5E-09\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS01870\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003epanB\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3-methyl-2-oxobutanoate hydroxymethyltransferase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e314\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e105\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.1E-08\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS01875\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003epanc\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003epantoate-beta-alanine ligase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e350\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e115\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.2E-08\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS01760\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ehypothetical protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e820\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e369\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e8.9E-08\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS02315\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ebasic amino acid ABC transporter substrate-binding protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e147\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.8E-07\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003efruK\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1-phosphofructokinase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1256\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e276\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.7E-06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00095\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePTS fructose transporter subunit IIC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1273\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e372\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.8E-06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00105\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eDeoR/GlpR transcriptional regulator\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1304\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e278\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e3.6E-06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"8\"\u003eL1: lactate-supplemented cells at growth phase\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"8\"\u003eC1: no-lactate-supplemented cells (controls) at growth phase\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe COG distribution of the DEGs at both the growth phase and the stationary phase was illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e. It revealed the potential genes related to the pathways and bioprocesses for the utilization of lactate for CA production. At the growth phase, predominant number of DEGs belong to the \u0026lsquo;inorganic ion transport and metabolism, [P]\u0026rsquo; group and the \u0026lsquo;carbohydrate transport and metabolism, [G]\u0026rsquo; group. While at stationary phase, most upregulated genes belong to the \u0026lsquo;carbohydrate transport and metabolism, [G]\u0026rsquo; group, which play important roles for the degradation and utilization of carbohydrate substrates [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. It was worth noting that a large number of DEGs (for both upregulated and downregulated ones) fall into the \u0026lsquo;function unknown, [S]\u0026rsquo; group.), meaning that their functions are unknown. This may be because CPB6 belongs to a novel species or clade (\u003cem\u003eClostridium\u003c/em\u003e cluster IV) of the family \u003cem\u003eRuminococcaceae\u003c/em\u003e, sharing low 16S rRNA sequence similarity (\u0026lt;\u0026thinsp;92.6%) with the other organisms in GenBank and RDP [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eCluster analysis of the DEGs between the culture with lactate supplementation and the control was showed in Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e. The results showed that the gene expression of the triplicate (a, b and c) of each sample demonstrated very similar expression patterns. According to Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e, L1 (with lactate supplementation at the growth phase) cluster was most closely with C1 (without lactate supplementation at the growth phase) cluster, indicating the presence of a small amount of DEGs caused by the addition of lactate at this phase. However, the distinct difference of gene expression was observed between the two cultures at the stationary phase (L2 vs C2), Further, venn diagram showed that 295 genes whose changes in expression patterns were substrate and/or growth dependent, of which 31 genes were substrate (lactate) dependent, 228 genes were growth dependent, and 36 genes were substrate-growth dependent (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003ea). Specifically, 11 and 20 lactate-dependent genes were significantly upregulated and downregulated, as well as 98 and 130 growth-dependent genes were significantly upregulated and downregulated, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eb). It was suggested that the differences in gene expression are stronger for stationary phase vs growth phase than for plus/minus lactate. Similar results was observed for \u003cem\u003eClostridium thermocellum\u003c/em\u003e, in which growth rate had stronger effects on gene expression than substrate type (insoluble cellulose vs soluble cellobiose) [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. These DEGs were described in detail in later section.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eExpression of glycolysis genes\u003c/h2\u003e \u003cp\u003eAn overview of the metabolic pathway in strain CPB6 and the expression levels of genes involved in key metabolic processes with their fold change (FC) were shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e and Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. Generally, in clostridia, glucose is converted into pyruvate via glycolysis, and the produced pyruvate is further converted into acetyl-CoA for the production of acetate and butyrate at acidogenic phase [\u003cspan additionalcitationids=\"CR24\" citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. In the present study, the expression of genes for glycolysis was detected in the CPB6 transcriptome, which reinforced its genome annotation [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Most glycolytic genes were expressed at a relatively high level (TPM\u0026thinsp;\u0026gt;\u0026thinsp;150) between the culture with lactate supplementation and the control, but there was no significant difference between them at the growth phase. Three glycolytic genes exhibited different expression patterns at the stationary phase. Gene encoding phosphofructokinase (PFK, B6259_RS06095) was significantly downregulated, while genes encoding glucose-1-phosphate adenylyltransferase (GlgC, B6259_RS09035) and 1, 4-alpha-glucan branching enzyme (GlgB, B6259_RS09040) were upregulated by 4.58 and 3.42-fold in the culture with lactate supplementation than in the control, respectively. GlgB and GlgC are typically associated with glycogen synthesis, why expression of these genes be affected by lactate supplementation remains unclear. Overall, the addition of lactate has little impact on the expression of glycolytic genes.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe differentially expressed genes within the important metabolic pathways in culture with/without lactate supplementation\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eFunctional description\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eGene_ID\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003eTPM of genes from culture with lactate supplementation\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003eTPM of genes from the Control\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003eRNA relative fold change (Treatment/Control)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e12\u0026nbsp;h\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e18\u0026nbsp;h\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e12\u0026nbsp;h\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e18\u0026nbsp;h\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e12\u0026nbsp;h\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e18\u0026nbsp;h\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eGlycolysis\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePTS-Glc-EIIA, PTS glucose transporter subunit IIA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09280\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e484\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e260\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e517\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e0.37\u003c/b\u003e\u003csup\u003e\u003cb\u003eb\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.62\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGlgC, glucose-1-phosphate adenylyltransferase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09035\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e153\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1323\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e236\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e241\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e4.58\u003c/b\u003e\u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGlgB, 1,4-alpha-glucan branching enzyme\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09040\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e194\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e745\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e236\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e201\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e3.42\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003esugar phosphate isomerase/epimerase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS06500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e181\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e175\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e150\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e233\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.88\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePGM, phosphoglucomutase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e189\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e127\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e113\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.80\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGPI, glucose-6-phosphate isomerase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS04825\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2015\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1789\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1833\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1818\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.12\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePFK, phosphofructokinase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS06095\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e426\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e580\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e516\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.23\u003c/b\u003e \u003csup\u003e\u003cb\u003eb\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eALDO, fructose-bisphosphate aldolase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00415\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e749\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e402\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e800\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e891\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.57\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTPI, triose-phosphate isomerase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09105\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e224\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e229\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e315\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e493\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.65\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.58\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGapA, glyceraldehyde phosphate dehydrogenase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09050\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e5322\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4284\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e4790\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e7732\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.70\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePGK, phosphoglycerate kinase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e523\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e524\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e705\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1029\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.65\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003egpmI, 2,3-bisphosphoglycerate-independent phosphoglycerate mutase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09110\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e203\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e284\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e469\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.55\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eENO, phosphopyruvate hydratase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS04810\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e65\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.46\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePK, pyruvate kinase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS02335\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e254\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e102\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e293\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e228\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.46\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCentral pyruvate metabolism\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePpdK, pyruvate phosphate dikinase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00120\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1301\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e823\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1163\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1535\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.69\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePfor, pyruvate: ferredoxin (flavodoxin) oxidoreductase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09135\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e4329\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4382\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2044\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1225\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e3.26\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePCK, phosphoenolpyruvate carboxykinase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09255\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e368\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e159\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e554\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1031\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.23\u003c/b\u003e \u003csup\u003e\u003cb\u003eb\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePflD, formate C-acetyltransferase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09900\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e188\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e107\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e471\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.54\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eADH, alcohol dehydrogenase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS03100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e116\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e163\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e159\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.84\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eIncomplete TCA cycle\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCS, citrate synthase, citrate lyase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS03360\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e936\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e187\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e543\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e642\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.39\u003c/b\u003e \u003csup\u003e\u003cb\u003eb\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eACO, aconitate hydratase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS05795\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e227\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e162\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e153\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e201\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.94\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIDH, isocitrate dehydrogenase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS05805\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e237\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e232\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e197\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e291\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.93\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFUM, fumarate hydratase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07270\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e310\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e186\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e260\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e437\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.49\u003c/b\u003e \u003csup\u003e\u003cb\u003eb\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePCK, phosphoenolpyruvate carboxykinase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09255\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e368\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e159\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e554\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1031\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.23\u003c/b\u003e \u003csup\u003e\u003cb\u003eb\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eHydrogen production\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHydE, [FeFe] hydrogenase H-cluster\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS02550\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e113\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e174\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e2.24\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHydF, [FeFe] hydrogenase H-cluster\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09690\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.17\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLactate fermentation pathway\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eD-ldh, D-lactate dehydrogenase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS06770\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e108\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.95\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eL-ldh, L-lactate dehydrogenase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09845\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e111\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e119\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e295\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.44\u003c/b\u003e \u003csup\u003e\u003cb\u003eb\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAcetate fermentation pathway\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePTA, phosphate acetyltransferase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07830\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e666\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e697\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e271\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e321\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e2.09\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e2.23\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eACK, acetate kinase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS03430\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e290\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e297\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e288\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e233\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.41\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eThe reverse β-oxidation pathway\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAtoB, acetyl-CoA C-acetyltransferase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS06365\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e5204\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e9909\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1224\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1077\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e3.45\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e6.31\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHBD, 3-hydroxybutyryl-CoA dehydrogenase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS06355\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6306\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e13975\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1418\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1022\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e3.49\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e8.59\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCRT, enoyl-CoA hydratase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS06360\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3434\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e7348\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e795\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e647\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e3.46\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e7.34\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBCD1, butyryl-CoA dehydrogenase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS01790\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3278\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3104\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3787\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e3014\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.19\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBCD2, butyryl-CoA dehydrogenase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS02600\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e313\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e4.49\u003c/b\u003e\u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEtfA, electron transfer flavoprotein subunit alpha\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS01785\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2657\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2968\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3175\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2572\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.31\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEtfB, electron transfer flavoprotein subunit beta\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS01780\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3996\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4830\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e4357\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e4169\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.31\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCAT, butyryl-CoA: acetate CoA-transferase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS06345\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e521\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1497\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e283\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e330\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e4.01\u003c/b\u003e\u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eFructose fermentation pathway\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePPF, 1-phosphofructokinase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e276\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2174\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1256\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e239\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e7.33\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eStarch and sucrose metabolism\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePYG, glycogen phosphorylase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00300\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e163\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e121\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e103\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.71\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMalQ, 4-alpha-glucanotransferase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07805\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e270\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e4.34\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePGM, Phosphoglucomutase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e189\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e127\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e113\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.80\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eEnergy conservation\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eenergy-coupling factor transporter ATPase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS02790\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e141\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e104\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e117\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e159\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.76\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eelectron transport complex protein RnfA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS06245\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e230\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e162\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e357\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e362\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.52\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSporulation\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003estage 0 sporulation protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00205\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e379\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e279\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e233\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e252\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.82\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003estage II sporulation protein D\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09065\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.29\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003estage III sporulation protein AD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS03910\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e126\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e54\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.27\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003estage IV sporulation protein A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS04975\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e65\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.58\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003estage V sporulation protein AC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09190\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.42\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003estage V sporulation protein AD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09195\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e69\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.57\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003estage V sporulation protein AE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e292\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e226\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e167\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.02\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003esporulation transcription factor Spo0A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS05505\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e127\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e115\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e106\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.94\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003esporulation transcriptional regulator SpoIIID\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS01550\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e213\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e188\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e140\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e207\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003esporulation protein YtfJ\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS04885\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e291\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e183\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e145\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e159\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.65\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTransporter genes\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eABC transporter permease\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00445\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e446\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e274\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e235\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e5.17\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.27\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003emetal ABC transporter\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00450\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e699\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e628\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e457\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e5.69\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.52\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eABC transporter permease\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS02670\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e296\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e130\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e441\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e387\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.40\u003c/b\u003e \u003csup\u003e\u003cb\u003eb\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eABC transporter permease\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS02665\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e180\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e258\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e231\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.48\u003c/b\u003e \u003csup\u003e\u003cb\u003eb\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ecarbohydrate ABC transporter permease\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e124\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e0.37\u003c/b\u003e \u003csup\u003e\u003cb\u003eb\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e3.51\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ecarbohydrate ABC transporter permease\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07905\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e744\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e12.71\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ecarbohydrate ABC transporter permease\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07810\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e229\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e5.48\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ecarbohydrate ABC transporter permease\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS02030\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e2.14\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003esugar ABC transporter permease\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07910\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1175\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e88\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e14.74\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003esugar ABC transporter permease\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS03335\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e401\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e5.61\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003esugar ABC transporter permease\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07815\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e197\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e4.34\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003esugar ABC transporter permease\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e135\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e113\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e0.30\u003c/b\u003e \u003csup\u003e\u003cb\u003eb\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e3.48\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eiron ABC transporter permease\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00320\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1278\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e10.05\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eABC transporter ATP-binding protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00440\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e504\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e277\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e239\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e5.25\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e1.39\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eABC transporter ATP-binding protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00325\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2032\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e11.14\u003c/b\u003e \u003csup\u003e\u003cb\u003ec\u003c/b\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eABC transporter ATP-binding protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS08900\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e153\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e682\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e233\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e214\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e3.13\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eABC transporter ATP-binding protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07940\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e190\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e259\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.42\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ecarbohydrate ABC transporter substrate-binding protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07915\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e216\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3434\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e203\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e103\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e14.51\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003emaltose ABC transporter substrate-binding protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS03345\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e501\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e7.65\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eABC transporter substrate-binding protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07820\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e372\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1913\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e451\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e344\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e4.63\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003esugar ABC transporter substrate-binding protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS02005\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.92\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e2.29\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003epeptide ABC transporter substrate-binding protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS08515\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e369\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e0.48\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.28\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003epeptide ABC transporter substrate-binding protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS02685\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1385\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e819\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1442\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2222\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.50\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eABC transporter ATP-binding protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS02660\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e238\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e119\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e369\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e320\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.45\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eABC transporter ATP-binding protein\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS07940\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e190\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e259\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e166\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.42\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePTS fructose transporter subunit IIC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00095\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e372\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2117\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1273\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e485\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e0.37\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e3.87\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePTS glucose transporter subunit IIA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS09280\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e484\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e260\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e517\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e0.37\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.62\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePTS β-glucoside transporter subunit IIABC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS01415\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e81\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e760\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e134\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e141\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.54\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e4.70\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePTS mannitol transporter subunit IICBA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS00370\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e89\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e2.34\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eferrous iron transport protein B\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eB6259_RS03880\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e471\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e389\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e531\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e150\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.81\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e2.72\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"8\"\u003ea, Data presented as mean of independent triplicates\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"8\"\u003eb, Significantly upregulated (FC\u0026thinsp;\u0026ge;\u0026thinsp;2.0, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05)\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"8\"\u003ec, Significantly downregulated (FC\u0026thinsp;\u0026le;\u0026thinsp;0.5, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05)\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eExpression of butyrate- and CA-producing genes\u003c/h2\u003e \u003cp\u003eThe bioproduction of CA is a well-known chain elongation process from acetate (C2) to caproate (C6) via the reverse β-oxidation pathway, in which an acetyl-CoA (from ethanol) unit is combined with another acetyl-CoA (from acetate), and consequently C2 is elongated to C4, and further C4 is elongated to C6 [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Thus, acetyl-CoA is a key intermediate of flux distribution for the chain elongation. The conversion of pyruvate into acetyl-CoA is mainly catalyzed by the pyruvate: ferredoxin (flavodoxin) oxidoreductase (Pfor) that is a flavodoxin- and NADPH-dependent enzyme [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Here, the Pfor gene (B6259_RS09135) maintained at very high expression level (TPM\u0026thinsp;\u0026gt;\u0026thinsp;4000) in the culture with lactate supplementation (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e), which were upregulated by 1.83- and 3.26-fold than that in the control in the growth and stationary phase, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e). High-level expression of the Pfor gene would result in increased acetyl-CoA, which provides the high amount of acetyl-CoA for chain elongation from acetate to butyrate and CA.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eKey enzymes involved in the butyrate formation include acetyl-CoA C-acetyltransferase (AtoB), 3-hydroxybutyryl-CoA dehydrogenase (HBD), enoyl-CoA hydratase (CRT), NAD-dependent butyryl-CoA dehydrogenase (BCD), Electron transfer flavoprotein (Etf) and butyryl-CoA: acetate CoA transferase (CAT) [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Here, genes encoding AtoB (B6259_RS06365), CRT (B6259_RS06360) and HBD (B6259_RS06355) were identified from the transcriptomes of strain CPB6, whose expression levels maintained at very high levels (TPM\u0026thinsp;\u0026gt;\u0026thinsp;3000) in the culture with lactate supplementation throughout the growth and stationary phases, and were upregulated by 3.5\u0026ndash;8.6 folds compared to the control. It suggested that the high-level expression of the three genes can be induced by supplemented lactate. In addition, two BCD genes (B6259_RS01790 and _RS02600) and EtfAB (B6259_RS01785 and _RS01780) responsible for the conversion of crotonyl-CoA to butyryl-CoA showed different expression profiles. B6259_RS01790 was expressed at a very high level (TPM\u0026thinsp;\u0026gt;\u0026thinsp;3000) throughout the fermentation phases, but showing no change in expression in the two cultures. B6259_RS02600 was expressed at relatively low level at the growth phase, but its expression was induced to high levels in the culture with lactate supplementation at the stationary phase, which was \u0026gt;\u0026thinsp;4.4-fold higher than that in the control. More research is needed to determine which BCD gene plays the key role in acidogenesis in the CPB6. EtfAB showed no significant change in the two cultures during the growth and stationary phases. A CAT gene (B6259_RS06345) was detected in the CPB6 transcriptome, and its expression was markedly upregulated by 4-fold in the culture with lactate supplementation than that in the control in the stationary phase, and kept at very high expression levels. CAT is key enzyme responsible for catalysing the last step of the butyrate formation [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. High-level expression of CAT gene should theoretically result in a high concentration of butyric acid in the culture with lactate supplementation. Nevertheless, significant accumulation of CA instead of butyric acid was observed in the CPB6 culture with lactate supplementation, suggesting that the CAT is likely more intent to transforming caproy-CoA to caproate instead of converting butyryl-CoA to butyrate. Up to now, little is known about the key functional genes responsible for CA synthesis from butyryl-CoA. Genes involved in butyrate synthesis via the reverse β-oxidation (e.g., AtoB, CRT, HBD, BCD and CAT) are assumed to have the function in the caproyl-CoA and CA formation [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. However, \u003cem\u003eClostridium\u003c/em\u003e sp. BPY5 and \u003cem\u003eC\u003c/em\u003e. \u003cem\u003etyrobutyricum\u003c/em\u003e, which contains these genes, only produce butyric acid instead of CA [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e], while \u003cem\u003eC\u003c/em\u003e. \u003cem\u003ekluyveri\u003c/em\u003e and \u003cem\u003eRuminococcaceae\u003c/em\u003e bacterium CPB6, which contain these genes, can further elongate butyric acid to CA [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. It suggested that there may be differences in structure and function between these genes from different organisms. Therefore, the further study needs to be performed to explore the functions of these genes in strain CPB6.\u003c/p\u003e \u003cp\u003eLactate is a major endproduct of glycolysis in the absence of oxygen [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Its formation or conversion requires lactate dehydrogenase (LDH) with the regeneration of NADH to NAD+. There are two LDH genes in the CPB6 genome [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e], encoding L-lactate dehydrogenase (L-LDH) and D-lactate dehydrogenase (D-LDH), respectively. In this study, the expression of LDH genes (L-LDH, B6259_RS09845; D-LDH, B6259_RS06770) were also detected in the transcripts of strain CPB6. However, the two genes showed relatively low expression levels in both the culture with lactate supplementation and the control, except that slightly higher expression level was observed in the control at the stationary phase (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). LDH catalyzes the reaction converts pyruvate to lactate or the reverse reaction that converts lactate to pyruvate [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. This conversion is essential in hypoxic and anaerobic conditions when ATP production by oxidative phosphorylation is disrupted. The recent study showed that lactate can be transformed into CA in either mixed microbiome [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], or in the pure culture where the conversion of lactate to acetyl-CoA coupling with the reverse β-oxidation is speculated to result into chain elongation [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Interestingly, the lactate supplementation in this study did not lead to increased expression levels of LDHs in either growth or stationary phases, indicating that the expression of LDHs might be uncoupled from the utilization of lactate. It warrants further investigation concerning the function of LDH in the conversion of lactate to CA in the CPB6 strain.\u003c/p\u003e \u003cp\u003eThe gene encoding phosphate acetyltransferase (PTA, B6259_RS07830), one important enzyme involved in acetate formation, was remarkably upregulated in the culture with lactate supplementation than in the control. However, the expression of acetate kinase (ACK, B6259_RS03430) showed no change in response to the addition of lactate. By including the production of H\u003csub\u003e2\u003c/sub\u003e and CO\u003csub\u003e2\u003c/sub\u003e into the loop, it could provide a whole picture for carbon balance for the substrate utilization and cell biomass production. Unfortunately, the production of H\u003csub\u003e2\u003c/sub\u003e and CO\u003csub\u003e2\u003c/sub\u003e was not monitored in this study. In the future studies, this should be taken into consideration for improvement.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eExpression of putative ABC transporter and sporulation genes\u003c/h2\u003e \u003cp\u003eStrain CPB6, affiliated with \u003cem\u003eClostridium\u003c/em\u003e cluster IV of the family \u003cem\u003eRuminococcaceae\u003c/em\u003e in the order of Clostridiales, is a spore-forming, obligate anaerobic bacterium that can produce CA from lactate (Zhu et al. 2017). As shown in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, sporulation genes showed similar expression patterns in both groups, e.g., \u003cem\u003espo\u003c/em\u003e0, \u003cem\u003espo\u003c/em\u003eIIID, \u003cem\u003espo\u003c/em\u003eV, \u003cem\u003espo\u003c/em\u003eYtfJ, were induced to high expression under both conditions (with or without lactate supplementation) at the growth and stationary phases, while \u003cem\u003espo\u003c/em\u003eIID, \u003cem\u003espo\u003c/em\u003eIIIAD, \u003cem\u003espo\u003c/em\u003eIVA, \u003cem\u003espo\u003c/em\u003eVAC, \u003cem\u003espo\u003c/em\u003eVAD \u003cem\u003espo\u003c/em\u003eVAE were expressed at low or moderate levels. Some bacteria, such as bacilli and clostridia, develop into highly resistant spores to protect their genome and cell from certain doom when living conditions become intolerable [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. It ensures bacterial survival under adverse environmental conditions. Sporulation in \u003cem\u003eClostridium\u003c/em\u003e spp. is ordinarily not triggered by starvation but by cessation of growth in the presence of excess carbon source or exposure to oxygen [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. The two most critical factors involved in the shift to solventogenesis, a decrease in external pH and accumulation of acidic fermentation products, are generally assumed to be associated with the initiation of sporulation in \u003cem\u003eClostridium\u003c/em\u003e spp., to some extent [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. But recent studies showed that the sporulation events were uncoupled from the induction of solventogenesis in \u003cem\u003eC. beijerinckii\u003c/em\u003e [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. In this study, the sporulation genes showed no significant difference between the culture with lactate supplementation and the control, indicating that the sporulation events are not associated with the production of CA in the CPB6 until the stationary phase. This may be because low concentrations of CA (1717\u0026nbsp;mg/L) are not sufficient to initiate sporulation for the CPB6. It will be investigated in the future whether high concentrations of CA trigger sporulation.\u003c/p\u003e \u003cp\u003eIn the transcriptome of strain CPB6, most genes encoding ATP-binding cassette (ABC) transporters and substrate-bind proteins (SBP) maintained at low expression levels in the control, but were induced to relatively high expression levels (particularly upregulated by 2\u0026ndash;14 folds at the stationary phase) in the culture with lactate supplementation. ABC transporters are ubiquitous membrane proteins that couple the transport of diverse substrates across cellular membranes to the hydrolysis of ATP [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. ABC transporters are generally divided into importers and exporters on the basis of the polarity of solute movement. ABC importers are found mostly in bacteria and are crucial in mediating the uptake of solutes including sugar, metal ions and vitamins [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. In the present study, most genes for ABC transporter and substrate-binding protein (SBP) were no significant change in the two cultures, except two ABC transporter genes (B6259_RS00445, B6259_RS00450), and one SBP gene (B6259_RS00440) which were upregulated by more than 2-fold in the culture with lactate supplementation than in the control. It was suggested that the three genes might be related to the intake and use of lactic acid. In addition, nine ABC transporter genes and six SBP genes were markedly upregulated at the stationary phase. Specially, B6259_RS07905, _RS07910, _RS00320, _RS00325 and B6259_RS07915 were increased over 10-fold in the culture with lactate supplementation than in the control, demonstrating that these genes are associated with the extrusion of CA from the cell and the maintenance of osmotic homeostasis in cytoplasm [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eIn addition, two phosphotransferase system (PTS) transporter genes (B6259_RS01415 and B6259_RS00370) and one ferrous iron transporter gene (B6259_RS03880) were upregulated by 2- to 4-fold in the culture with lactate supplementation than in the control. PTS is a multiple-component carbohydrate uptake system that drives specific saccharides across the bacterial inner membrane while simultaneously catalyzing sugar phosphorylation [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. Five distinct subfamilies of proteins related to PTS have been identified within the glucose superfamily: the lactose family, the glucose family, the β-glucoside family, the mannitol family, and the fructose family [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. In this study, four PTS transporter genes were detected in the transcriptome of strain CPB6, including PTS fructose transporter subunit IIC (B6259_RS00095), PTS glucose transporter subunit IIA (B6259_RS09280), PTS β-glucoside transporter subunit IIABC (B6259_RS01415), and PTS mannitol transporter subunit IICBA (B6259_RS00370). Genes encoding PTS fructose and glucose transporters were highly expressed under both conditions, but the two genes were significantly downregulated at the growth phase in the culture with lactate supplementation than in the control, indicating that PTS transporter-mediated sugar transport in membrane vesicles in CPB6 is inhibited by the lactate supplementation. However, at the stationary phase, genes encoding PTS fructose, β-glucoside and mannitol transporters were all strikingly upregulated.\u003c/p\u003e \u003cp\u003eOverall, the addition of lactate caused distinct changes in gene expression of the CPB6 strain, especially at stationary phase. The detailed mechanism remains to be further studied.\u003c/p\u003e \u003c/div\u003e "},{"header":"Conclussions","content":" \u003cp\u003eThis study showed that lactate supplementation induced earlier CA production, higher CA titer and productivity, but had little impact on cell growth, suggesting that the supplemented lactate is more involved in secondary metabolism (e.g., the production of CA) than the carbon assimilation. The gene transcriptional profiles based on RNA-Seq demonstrated that supplemented lactate promoted CA production by altering the expression patterns of genes involved in crucial metabolic pathways. Specifically, the Pfor gene involved in central pyruvate metabolism, 5 genes (AtoB, HBD, CRT, BCD and CAT) involved in the reverse β-oxidation pathway, 11 genes encoding ABC transporter, 6 SBP genes, and 4 PTS transporter genes showed sharp upregulation in response to the addition of lactate. The findings presented herein provide unique insights into the metabolic effects of lactate on CA production at the gene regulation level.\u003c/p\u003e "},{"header":"Methods","content":" \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eMicroorganisms, media and fermentation experiment\u003c/h2\u003e \u003cp\u003e \u003cem\u003eRuminococcaceae\u003c/em\u003e bacterium CPB6 (GDMCC No.60133) was routinely cultured at 37 \u003csup\u003eo\u003c/sup\u003eC anaerobically in a modified tryptone-glucose-yeast extract (mTGY) medium containing the following compounds (g L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, pH 6.0): tryptone, 5.0; glucose, 2.0; yeast extract, 5.0; sodium acetate, 3.5; K\u003csub\u003e2\u003c/sub\u003eHPO\u003csub\u003e4\u003c/sub\u003e\u0026middot;3H\u003csub\u003e2\u003c/sub\u003eO, 0.41; KH\u003csub\u003e2\u003c/sub\u003ePO\u003csub\u003e4\u003c/sub\u003e, 0.23; NH\u003csub\u003e4\u003c/sub\u003eCl, 0.25; MgSO\u003csub\u003e4\u003c/sub\u003e\u0026middot;7H\u003csub\u003e2\u003c/sub\u003eO, 0.20; NaHCO\u003csub\u003e3\u003c/sub\u003e, 2.5; L-cysteine, 0.25; Na\u003csub\u003e2\u003c/sub\u003eS\u0026middot;9H\u003csub\u003e2\u003c/sub\u003eO, 0.25; resazurin, 0.0005; and 1\u0026nbsp;ml of trace element solution SL-10 and 1\u0026nbsp;ml of vitamin solution [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. The suspension of activated strain CPB6 was inoculated with a 5% ratio into the same medium as described above and incubated for 12\u0026ndash;15\u0026nbsp;h until the OD\u003csub\u003e600\u003c/sub\u003e of the culture reached 0.8-1.0. Then the culture would be used as the seed inoculum (5% ratio, v/v) for batch experiments. To investigate the effect of lactate on cell growth and CA production, 5\u0026nbsp;g L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e sodium lactate was supplemented into the modified TGY medium (mTGYL). Batch experiments were performed in 250\u0026nbsp;mL serum bottles containing 100\u0026nbsp;mL of modified mTGY or mTGYL media. The headspace of the bottle was filled with highly pure N\u003csub\u003e2\u003c/sub\u003e. Each fermentation was performed in triplicate. The fermentation was carried out at 37 \u003csup\u003eo\u003c/sup\u003eC for 24\u0026nbsp;h in an E500 anaerobic workstation (Gene Science, USA) under N\u003csub\u003e2\u003c/sub\u003e: CO\u003csub\u003e2\u003c/sub\u003e: H\u003csub\u003e2\u003c/sub\u003e (volume ratio of 80:10:10) atmosphere.\u003c/p\u003e \u003cp\u003eSamples were taken at specific times and processed for cell concentration determination and HPLC analysis. Samples for RNA isolation were taken at the growth and stationary phases of the cell growth.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eAnalytical methods\u003c/h2\u003e \u003cp\u003eCulture growth was monitored by measuring the optical density at 600\u0026nbsp;nm (OD\u003csub\u003e600\u003c/sub\u003e) using a TU-1810 UV/Vis Spectrophotometer (Puxi Instrument Co. Ltd., Beijing). Lactic acid, acetic acid, butyric acid, and caproic acid were quantified using an HPLC system (Agilent 1260 Infinity, USA) equipped with a differential refraction detector (RID) and a Agilent Hi-Plex H column (300\u0026thinsp;\u0026times;\u0026thinsp;6.5\u0026nbsp;mm) following the procedure as previously described [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eRNA isolation, library construction, and sequencing\u003c/h2\u003e \u003cp\u003eIn preparation for RNA isolation, 10\u0026nbsp;ml cell culture was harvested at each time point, and centrifuged at 8,000\u0026thinsp;\u0026times;\u0026thinsp;g for 10\u0026nbsp;min at 4\u0026nbsp;\u0026deg;C. Cells were then frozen in liquid nitrogen prior to storage at -80\u0026nbsp;\u0026deg;C. The RNA was extracted and purified using a RNA extraction kit (DP430, Tiangen Biotech, Beijing, China) following the manufacturer's protocol. RNA quality and quantity were characterized using a NanoDrop2000 (NanoDrop Technologies, Wilmington, DE), agarose gel electrophoresis (RNA integrity detection) and Agilent 2100 (RIN value measurement). Only the RNA samples with high-quality (\u0026ge;\u0026thinsp;5\u0026nbsp;\u0026micro;g; \u0026ge;200\u0026nbsp;ng/\u0026micro;L; OD260/280\u0026thinsp;=\u0026thinsp;1.8\u0026thinsp;~\u0026thinsp;2.2) were used for the cDNA library construction and sequencing. Before library construction, rRNAs were removed with the Ribo-Zero rRNA Removal Kit (Epicentre, San Diego, CA) following the manufacturer\u0026rsquo;s protocol. The enriched mRNA was fragmented into short fragments (approximately 200\u0026nbsp;bp) randomly using metal ions. Then the first strand cDNA was synthesized using the random hexamer-primer with the mRNA fragment as the template. After synthesizing the second strand cDNA using DNA polymerase I and RNase H, double-stranded cDNA was further end repaired, A-tailed, and indexed adapters ligated. The final cDNA library was constructed using the mRNA-Seq library construction kit (Illumina Inc., San Diego, CA), and then sequenced by Illumina platform (Illumina Inc., San Diego, CA) with 2\u0026thinsp;\u0026times;\u0026thinsp;150\u0026nbsp;bp.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eRNA-Seq data analysis\u003c/h2\u003e \u003cp\u003eHigh quality reads in each sample were aligned to the CPB6 genome using Bowtie2, and those that did not align uniquely to the genome were discarded using the default quality parameters. Each base was assigned a value based on the number of mapped sequence coverage. Gene expression levels were defined using the number of transcripts per million (TPM), which is proportional to the quantity of cDNA fragments derived from the gene transcripts. The quantitative gene expression values between samples were identified by calculating the number of unambiguous tags for each gene and then normalizing this to TPM, which was calculated following the method reported by Parto et al., [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. The gene expression results were visualized using heatmap plots.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eSignificant differences of the gene expression between the culture from fermentation with lactate supplementation vs. the control were determined using ANOVA in R software (version 3.5.2). TPM values were first transformed to log10-scale. The log10-transformed TPM values were then properly centered for better representation of the data using the heatmap plots. Fold changes (FCs) as the ratio of the TPM values were calculated following the method reported by Love et al. [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e], and were used to compare the differentially expressed genes (DEGs) between the culture from fermentation with lactate supplementation and the control.\u003c/p\u003e \u003c/div\u003e "},{"header":"Declarations","content":" \u003cp\u003e \u003ch2\u003eAvailability of data and materials\u003c/h2\u003e \u003cp\u003eThe RNA-Seq sequencing data have been deposited in the NCBI Sequence Read Archive (SRA) under the accession number PRJNA564589\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003ch2\u003eEthics approval and consent to participate\u003c/h2\u003e \u003cp\u003eNot applicable.\u003c/p\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eCompeting interests\u003c/strong\u003e \u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e \u003c/p\u003e \u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003eThis work has been supported by the Natural Science Foundation of China (31770090), the Open-foundation project of CAS Key Laboratory of Environmental and Applied Microbiology (KLCAS-2017-01).\u003c/p\u003e \u003ch2\u003eAuthors\u0026rsquo; contributions\u003c/h2\u003e \u003cp\u003eYT planned and analyzed the data, and wrote the manuscript. SL performed the experiments. YW participated in the planning and revision of manuscript, HJ participated in analysis of the data. CW and XH participated in the experiments. All authors discussed the results, read and approved the final manuscript.\u003c/p\u003e \u003ch2\u003eAcknowledgements\u003c/h2\u003e \u003cp\u003eWe thank Dr. Jun Feng for their assistance and advice for the study.\u003c/p\u003e "},{"header":"References","content":"\u003col\u003e\u003cli\u003e \u003cspan\u003eRoghair M, Liu Y, Adiatma JC, Weusthuis RA, Bruins ME, Buisman CJN, Strik D. Effect of n-Caproate Concentration on Chain Elongation and Competing Processes. ACS Sustain Chem Eng. 2018;6(6):7499\u0026ndash;506.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eNzeteu CO, Trego AC, Abram F, O'Flaherty V. Reproducible, high-yielding, biological caproate production from food waste using a single-phase anaerobic reactor system. 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Structural insight into the PTS sugar transporter EIIC. Biochim Biophys Acta. 2015;1850(3):577\u0026ndash;85.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eNguyen TX, Yen MR, Barabote RD, Saier MH Jr. Topological predictions for integral membrane permeases of the phosphoenolpyruvate:sugar phosphotransferase system. J Mol Microbiol Biotechnol. 2006;11(6):345\u0026ndash;60.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003ePatro R, Mount SM, Kingsford C. Sailfish enables alignment-free isoform quantification from RNA-seq reads using lightweight algorithms. Nat Biotechnol. 2014;32(5):462-U174.\u003c/span\u003e \u003c/li\u003e \u003cli\u003e \u003cspan\u003eLove MI, Huber W, Anders S. \u003cb\u003eModerated estimation of fold change and dispersion for RNA-seq data with DESeq2\u003c/b\u003e. \u003cem\u003eGenome Biol\u003c/em\u003e 2014, 15(12).\u003c/span\u003e \u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-genomics","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"gics","sideBox":"Learn more about [BMC Genomics](http://bmcgenomics.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/gics","title":"BMC Genomics","twitterHandle":"#BMCGenomics","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"n-Caproic acid, Lactate, Chain elongation, Transcriptome, RNA-Seq","lastPublishedDoi":"10.21203/rs.3.rs-76088/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-76088/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003e \u003cem\u003en\u003c/em\u003e-Caproic acid (CA) is gaining increased attention due to its high value as a chemical feedstock. \u003cem\u003eRuminococcaceae\u003c/em\u003e bacterium strain CPB6 is an anaerobic mesophilic bacterium that is highly prolific in its ability to perform chain elongation of lactate to CA. However, little is known about the genome-wide transcriptional analysis of strain CPB6 for CA production triggered by the supplementation of exogenous lactate.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eIn this study, 0.5% lactate was supplemented into the fermentation with \u003cem\u003eRuminococcaceae\u003c/em\u003e bacterium CPB6 for CA production. Compared to the control (without lactate supplementation), lactate supplementation led to earlier CA production and higher final CA titer and productivity. Transcriptional analysis was carried out using RNA-Seq for the culture with and without lactate supplementation (two groups) at growth and stationary phases, respectively. It has been indicated that 295 genes whose changes in expression patterns were substrate and/or growth dependent. These genes cover crucial functional categories. Specifically, 5 genes responsible for the reverse β-oxidation pathway, 11 genes encoding ATP-binding cassette (ABC) transporters, 6 genes encoding substrate-binding protein (SBP) and 4 genes encoding phosphotransferase system (PTS) transporters were strikingly upregulated in response to the addition of lactate. These genes would be candidates for future studies aiming at understanding the regulatory mechanism of lactate conversion into CA, as well as for the improvement of CA production in strain CPB6.\u003c/p\u003e\u003ch2\u003eConclusions\u003c/h2\u003e \u003cp\u003eThis study suggested that lactate supplementation can promote CA production by altering the expression patterns of genes involved in the essential metabolic pathways, such as central pyruvate metabolism, the reverse β-oxidation pathway, ABC and PTS transports. The findings presented herein reveal unique insights into the biomolecular effects of lactate on CA production at the gene transcriptional level.\u003c/p\u003e","manuscriptTitle":"Genome-wide transcriptomic analysis of n-caproic acid production in Ruminococcaceae bacterium CPB6 with lactate supplementation","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2020-09-22 15:45:30","doi":"10.21203/rs.3.rs-76088/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revision","date":"2021-01-28T00:00:00+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2021-01-05T00:00:00+00:00","index":2,"fulltext":"Recommendation: Reviewer's comments unavailable due to the journal's policy.\n"},{"type":"editorInvitedReview","content":"","date":"2021-01-05T00:00:00+00:00","index":3,"fulltext":"Recommendation: Reviewer's comments unavailable due to the journal's policy.\n"},{"type":"reviewerAgreed","content":"","date":"2020-12-14T01:00:00+00:00","index":3,"fulltext":""},{"type":"reviewerAgreed","content":"","date":"2020-12-14T00:00:00+00:00","index":2,"fulltext":""},{"type":"editorInvitedReview","content":"","date":"2020-12-06T00:00:00+00:00","index":1,"fulltext":"Recommendation: Reviewer's comments unavailable due to the journal's policy.\n"},{"type":"reviewersInvited","content":"","date":"2020-10-23T12:00:00+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2020-10-23T12:00:00+00:00","index":1,"fulltext":""},{"type":"editorAssigned","content":"","date":"2020-09-15T12:00:00+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2020-09-14T12:00:00+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2020-09-14T12:00:00+00:00","index":"","fulltext":""},{"type":"submitted","content":"","date":"2020-09-10T12:00:00+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-genomics","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"gics","sideBox":"Learn more about [BMC Genomics](http://bmcgenomics.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/gics","title":"BMC Genomics","twitterHandle":"#BMCGenomics","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"53ca9c6a-2e55-420a-93f1-33ead3c5343d","owner":[],"postedDate":"September 22nd, 2020","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":576785,"name":"Laboratory Diagnostics"}],"tags":[],"updatedAt":"2020-09-22T15:45:30+00:00","versionOfRecord":[],"versionCreatedAt":"2020-09-22 15:45:30","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-76088","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-76088","identity":"rs-76088","version":["v1"]},"buildId":"rHA-KDH7Qsr4HCuvH75dn","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}