Craterilacuimicrobium aquaticum gen. nov. sp. nov., a novel member of the order Neisseriales isolated from crater lake

Craterilacuimicrobium aquaticum gen. nov. sp. nov., a novel member of the order Neisseriales isolated from crater lake | 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 Craterilacuimicrobium aquaticum gen. nov. sp. nov., a novel member of the order Neisseriales isolated from crater lake You-Jun Liao, Xuan Zhang, Zi-Xuan Liu, Ai-Hua Li This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7595832/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 9 You are reading this latest preprint version Abstract Strain TC5R-5 T , a white rod-shaped bacterium, was isolated from Aershan Tianchi crater lake in northern China. It is Gram-stain-negative, catalase- and oxidase- positive. Optimal growth occurs at 20–30°C, pH of 6.0–7.0 and in the presence of 0-0.5% (w/v) NaCl. Based on 16S rRNA gene sequences analysis, strain TC5R-5 T exhibits the highest similarity (94.3%) to Chromobacterium alkanivorans IITR71 T . The genomic DNA G + C content of strain TC5R-5 T is 51.4% according to the genome sequence. The predominant cellular fatty acids are summed feature 3 (comprising C 16:1 ω 7 c /C 16:1 ω 6 c ) and C 16:0 . The predominant quinone is ubiquinone Q-8, with Q-7 present in moderate amounts. The polar lipid profile comprises phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), an unidentified aminophospholipid (APL), and an unidentified phospholipid (PL). Through polyphasic taxonomy and molecular phylogenetic analysis, strain TC5R-5 T is proposed to represent a novel species of a new genus of the family Chromobacteriaceae within the order Neisseriales , for which the name Craterilacuimicrobium aquaticum gen. nov. sp. nov. is proposed. The type strain is TC5R-5 T (= CGMCC 1.17058 T = KCTC 72736 T ). crater lake Craterilacuimicrobium Da Hinggan mountain Neisseriales Figures Figure 1 Figure 2 Figure 3 Introduction The order Neisseriales was reorganized by Adeolu et al . in 2013, dividing it into the families Neisseriaceae and Chromobacteriaceae based on phylogenomic and molecular signatures [Adeolu and Gupta, 2013 ]. Chen et al. classified 37 genera within the order Neisseriales , encompassing 122 species, into two revised families Neisseriaceae and Chromobacteriaceae , along with three new families namely Aquaspirillaceae fam. nov., Chitinibacteraceae fam. nov., and Leeiaceae fam. nov. [Chen et al., 2021 ]. The family Chromobacteriaceae contained the type genus Chromobacterium [De Ley et al., 1978 ] and other genera like Aquitalea [Lau et al., 2006 ], Craterilacuibacter [Liu et al., 2020 ], Crenobacter [Dong et al., 2015 ], Gulbenkiania [Vaz-Moreira et al., 2007 ], Paludibacterium [Kwon et al., 2008 ], Pseudogulbenkiania [Lin et al., 2008 ] and Vogesella [Grimes et al., 1997 ]. Strains of these genera were predominantly isolated from aqueous and soil habitats, such as soil, rice field and water samples. Aquitalea magnusonii [Lau et al., 2006 ], Gulbenkiania indica [Jyoti et al., 2010 ], Gulbenkiania mobilis [Vaz-Moreira et al., 2007 ], Vogesella lacus [Chou et al., 2009 ], and Pseudogulbenkiania subflava [Lin et al., 2008 ] were isolated from various aquatic environments, such as humic lake, sulfur spring, cold spring and pond. During the exploration of bacterial diversity of the Aershan Tianchi crater lake, strain TC5R-5 T was isolated from a water sample. Due to its low 16S rRNA similarity (94.3%) with the currently valid published species, a series of experiments were conducted to determine the taxonomic status of strain TC5R-5 T . Using polyphasic taxonomy, strain TC5R-5 T was classified as a novel species of a new genus within the order Neisseriales. Despite minor topological discrepancies among phylogenomic trees generated through different analytical methods, the collective evidence strongly supports its provisional assignment to the family Chromobacteriaceae as the most coherent and biologically plausible placement. Material and Methods Strain Isolation Strain TC5R-5 T was isolated from water samples collected from the surface water at a depth of 0.5 m in Aershan Tianchi crater lake, September 2015, located on Da Hinggan Ling Mountain in northern China (120.41°E, 47.32°N). The water temperature was 12°C. The samples were diluted from 10 − 1 to 10 − 5 gradient with sterile 0.85% NaCl (w/v) and cultured on R2A agar medium (Difco) at 25°C for 7 days. Single colonies were picked up and purified by streaking on R2A plate once again. The strains were preserved by lyophilization and in liquid nitrogen, respectively. 16S rRNA Gene Sequencing and Phylogenetic Analysis After being cultivated on R2A agar at 25℃, the genomic DNA of strain TC5R-5 T was extracted using the Genomic DNA Rapid Isolation Kit for bacterial cells (BioDev-Tech. Beijing, China) and the 16S rRNA gene of strain TC5R-5 T was amplified with the universal primers 27F and 1492R [Lane and Goodfellow, 1991 ]. An almost complete 16S rRNA gene sequence were obtained and analysed using the GenBank databases ( https://blast.ncbi.nlm.nih.gov/Blast.cgi ) and EzBioCloud ( https://www.ezbiocloud.net/ ) [Yoon et al., 2017 ]. Sequences of related taxa were downloaded and aligned using Clustal W [Larkin et al., 2007 ]. Phylogenetic trees were constructed using neighbor-joining, maximum-likelihood and maximum-parsimony methods in MEGA7.0 [Felsenstein, 1981 ; Fitch, 1971 ; Saitou and Nei, 1987 ; Tamura et al., 2011 ]. The evolutionary distance matrices were calculated using Kimura’s two parameter model [Kimura, 1980 ], and bootstrap values were determined based on 1000 replications [Felsenstein, 1985 ]. Genome Features and Phylogenomic Analysis The genome of strain TC5R-5 T was sequenced using the Illumina HiSeq X platform at Majorbio Co., Ltd. The qualified samples were pair-end sequenced, and raw sequencing data were acquired. Quality control used SeqPrep v0.20.0, the genome sequence was de novo assembled using SOAP v2.04. Contigs exceeding 400 bp in length were retained for gene prediction. CDS prediction was carried out using Glimmer v3.02 ( http://ccb.jhu.edu/software/glimmer/index.shtml ) and GeneMarkS v4.3 ( http://topaz.gatech.edu/GeneMark ). The rRNA and tRNA were predicted using Barrnap v0.8; ( https://github.com/tseemann/barrnap/ ) and tRNA-scan-SE v2.0 ( http://trna.ucsc.edu/software/ ), respectively. Average Nucleotide Identity (ANI) was determined through the OrthoANI algorithm on Ezbiocloud [Yoon et al., 2017 ]. The dDDH values were calculated using the TYGS with the GBDP method [Meier-Kolthoff and Göker, 2019 ]. Amino acid identity (AAI) was calculated using an AAI calculator ( http://enve-omics.ce . gatech.edu/aai/index) [Luo et al., 2014 ]. The ANI, dDDH and AAI values were compared with the thresholds established in literature [Chun et al., 2018 ; Richter and Rosselló-Móra, 2009 ]. The genes were classified into subsystems and clusters of orthologous groups (COGs) using the Rapid Annotation using Subsystem Technology (RAST) server [Aziz et al., 2008 ]. Genomic annotation was carried out using the NR ( ftp://ftp.ncbi.nlm.nih.gov/blast/db/ ), KEGG ( http://www.genome.jp/kegg/ ) and eggNOG ( http://eggnogdb.embl.de/#/app/home ) algorithms, respectively. The software antiSMASH 6.0 was utilized to predict the genes responsible for the secondary metabolites [Kai et al., 2021 ]. To confirmed the phylogenetic position of strain TC5R-5 T , three genome-based phylogenetic trees were reconstructed using different methods: one used the TYGS platform [Meier-Kolthoff and Göker, 2019 ], another was reconstructed with IQ-TREE v2.1.2 and based on single copy genes using MFP as the model and 1000 ultrafast bootstrap repeats [Minh et al., 2020 ], and the third phylogenomic tree was generated based on core genes using UBGC2.0 on EzBiocloud. Morphological and Physiological Analyses The morphological and physiological characteristics were determined through the following experiments. Cell morphology was observed using phase-contrast microscope and transmission electron microscope (JEM1400; JEOL) after 2 days of growth on R2A agar at 25℃ [Winey et al., 2014 ]. Cells were stained with uranium acetate before observation. Motility was assessed by inoculating strain TC5R-5 T into semi-solid R2A agar medium (0.5% agar) and observing its growth trace in the medium. A single colony of strain TC5R-5 T was circular, smooth, and white-colored. Cells were 0.8–0.9 µm in width and 3.0–4.0 µm in length. The optimal growth conditions of strain TC5R-5 T , including temperature range, pH and NaCl concentrations were confirmed as described below. Strain TC5R-5 T was cultured in R2A broth with NaCl concentrations ranging from 0 to 5.0% (w/v, at intervals of 0.5%). The pH variations (4.0, 5.0, 6.0, 6.5, 7.0, 7.5, 8.0, 9.0, 10.0) was achieved using specific buffers: 100mM acetate buffer (for pH 4.0–5.0), 100mM phosphate buffer (pH 6.0–8.0) or 100mM NaHCO 3 /Na 2 CO 3 buffer (pH 9.0–10.0). Growth at different temperatures (4, 15, 20, 25, 30, 35, 37, 40 and 45°C) was evaluated in R2A broth under the optimal NaCl concentration. Gram-staining, anaerobic growth and DNase activity were measured following previously described methods [Gerhardt et al., 1994 ]. The catalase and oxidase activities were assessed using 3% H 2 O 2 solution and 1% tetra-methyl- p -phenylenediamine (bioMérieux), respectively. Strain TC5R-5 T and its related reference strains, such as Chromobacterium alkanivorans IITR-71 T , Chromobacterium violaceum ATCC 12472 T , Aquitalea magnusonii TRO-001DR8 T , Paludibacterium yongneupense 5YN8-15 T , Pseudogulbenkiania subflava DSM 22618 T and Gulbenkiania mobilis E4FC31 T , were cultured on R2A agar at 25°C for 2 days for comparison. Craterilacusibacter sinensis B2N2-7 T was cultivated in PYG medium. Enzymatic and physiological activities were determined using a set of enzymatic tests API ZYM and 20NE (BioMérieux), and acids production was tested using API 50CH (BioMérieux) strips. Carbon sources utilization was measured using the GEN III MicroPlate system (Biolog) following the manufacturer’s protocol. Chemotaxonomic Analyses Strain TC5R-5 T was cultured in R2A broth at 25°C with 180 rpm for 48h and harvested during the late exponential phase. Cells were collected for further analysis of fatty acids, respiratory quinones, and polar lipids. Fatty acids were extracted following the standard protocol by Miller and Kuykendall and analysed using the Sherlock Microbial Identification System (MIDI) [Miller, 1982 ; Sasser, 1990 ]. The collected cells were freeze-dried for chemotaxonomic analysis. Polar lipids were extracted as per the protocol outlined by Komagata and Suzuki ( 1987 ) [Komagata and Suzuki, 1987 ] and separated using two-dimensional silica gel thin layer chromatography. The first direction involved a mixture of chloroform: methanol:water (65:25:4, v/v/v), while the second direction used a mixture of chloroform : acetic acid : methanol : water (80:15:12:4, v/v/v/v). Finally, molybdatophosphoric acid, molybdenum blue, ninhydrin and anisaldehyde reagent were utilized to detect total polar lipid, phospholipids, aminolipids, and glycolipids [Minnikin et al., 1993 ]. The specific components of respiratory quinones were extracted, separated, purified and analysed by HPLC following the previously described approach [ Collins, 1985 ; Wu et al., 1989 ]. Results and Discussion Phylogenetic and phylogenomic analysis Based on 16S rRNA gene sequence comparisons, strain TC5R-5 T exhibited the highest similarity (94.3%) to Chromobacterium alkanivorans IITR-71 T [Bajaj et al., 2016 ]. Phylogenetic analysis using the maximum-likelihood method with the GTR + G + I nucleotide substitution model placed strain TC5R-5 T within a clade containing the genera Chromobacterium , Aquitalea , Gulbenkiania , and Pseudogulbenkiania (Fig. 1 ). Consistent with this, phylogenetic trees reconstructed using neighbor-joining and maximum-parsimony methods also supported the close affiliation of strain TC5R-5 T with members of the family Chromobacteriaceae (order Neisseriales ). The topological structures of the neighbor-joining and maximum-parsimony trees were congruent with that of the maximum-likelihood tree (see Supplementary Material Figure S1 ). To further clarify the phylogenetic position of strain TC5R-5T, phylogenomic trees were reconstructed using TYGS, IQ-TREE, and UBGC. The maximum-likelihood tree generated with IQ-TREE v2.1.2 under the MFP model and 1000 ultrafast bootstrap replicates, with selected species from Neisseriales and Betaproteobacteria as outgroups, placed strain TC5R-5 T within a cluster comprising Chromobacterium species. This branch was neighbored by the genera Aquitalea , Gulbenkiania , Paludibacterium , and Pseudogulbenkiania , all belonging to the family Chromobacteriaceae (Fig. 2 a). A similar phylogenetic placement was supported by the bacterial core gene-based tree (Fig. 2 b), in which strain TC5R-5 T reliably grouped with members of the Chromobacteriaceae , including species of Paludibacterium , Aquitalea , Chromobacterium , Vogesella , and Gulbenkiania , among others. These results suggest that strain TC5R-5 T may represent a novel species within this family. In contrast, the TYGS-based phylogenomic tree revealed a divergent topology (Fig. 2 c). Here, strain TC5R-5 T clustered with ‘ Halomonas humidisoli ’, Vreelandella glaciei , and Andreprevotia sp . However, based on physiological and ecological characteristics, the strain shows no clear affiliation with Halomonas species. Notably, it shares certain similarities with Andreprevotia chitinilytica JS11-7 T , a member of the family Chitinibacteraceae . The ANI and dDDH values between strain TC5R-5 T and its closest relative, Chromobacterium alkanivorans IITR-71 T , were 73.5% and 13.6%, respectively—both well below the established species demarcation thresholds of 95–96% for ANI and 70% for dDDH. Additionally, the AAI value between the two strains was 52.8%, which fell beneath the proposed genus boundary range of 60–80% [Luo et al., 2014 ]. To further evaluate its taxonomic position, ANI values were compared between strain TC5R-5 T and several reference strains: the phylogenetically closest relatives within the Chromobacteriaceae (based on 16S rRNA and IQ-TREE/UBGC phylogenomic trees), Chromobacterium alkanivorans IITR-71 T and Aquitalea palustris CCM7557 T , as well as the top hits from the TYGS-based tree, including Andreprevotia chitinilytica DSM 18519 T , Halomonas humidisoli WN018 T , and Vreelandella glaciei DD39 T (Table S1 ). The ANI values between strain TC5R-5 T and the Chromobacteriaceae members C. alkanivorans IITR-71 T and A. palustris CCM7557 T (73.48%, 72.49%) were notably higher than those with A. chitinilytica DSM 18519 T , H. humidisoli WN018 T , and V. glaciei DD39 T (68.49%, 64.26%, 64.73%). These comparisons were also supported by substantially greater genome coverage (26.78% and 25.03% vs. 9.92%, 5.01%, and 4.91%, respectively). Together, these results corroborate that strain TC5R-5 T is phylogenetically more closely affiliated with the Chromobacteriaceae . In summary, integrated phylogenetic analyses based on both 16S rRNA gene and whole-genome sequences support the classification of strain TC5R-5 T as a novel species of a new genus within the order Neisseriales . Although some discrepancies were observed among phylogenomic trees constructed using different methods, current evidence indicates that its placement within the family Chromobacteriaceae is the most appropriate and rational provisional assignment. Genome Features and Biosynthetic Gene Cluster The complete genome of strain TC5R-5 T comprises 3.44 Mb with a total of 3408 genes, including 3274 protein-coding sequences (CDS). The genomic DNA G + C content, derived from sequencing data, was determined to be 51.4%. A comparative summary of genomic features between strain TC5R-5 T and its phylogenetic relatives is provided in Table 1 . Annotation via RAST revealed that 29% of the chromosomal genes could be assigned to functional categories, distributed across 291 subsystems within 27 broader functional groups. The most abundant subsystems were ‘Amino Acids and Derivatives’, ‘Protein Metabolism’, and ‘Cofactors, Vitamins, Prosthetic Groups, Pigments’ (Figure S2). AntiSMASH analysis identified a biosynthetic gene cluster (BGC) located on scaffold 7, spanning 24,974 bp, which showed high similarity to the rhizomide BGC from Paraburkholderia rhizoxinica HKI 454 (NC_014718.1; Figure S3). According to MIBiG annotation, this cluster is predicted to encode a nonribosomal peptide synthetase (NRPS) consisting of seven modules and containing a starter condensation domain, suggesting the potential production of a lipopeptide-class compound. Table 1 Comparison of genome features between strain TC5R-5 T and the type species of genus in the family Strains: 1. TC5R-5 T (JACCFC000000000); 2. Chromobacterium alkanivorans IITR-71 T (JAFHKL000000000); 3. Chromobacterium violaceum ATCC 12472 T (AE016825); 4. Aquitalea magnusonii DSM 25134 T (QJKC00000000); 5. Paludibacterium yongneupense 5YN8-15 T (AUGZ00000000); 6. Pseudogulbenkiania subflava DSM 22618 T (FXAG00000000); 7. Gulbenkiania mobilis E4FC31 T (SMDA00000000); 8. Craterilacuibacter sinensis B2N2-7 T (WSSB00000000); 9. Crenobacter luteus YIM 78141 T (SLXG00000000); 10. Vogesella indigofera DSM 3303 T (RBID00000000). 1 2 3 4 5 6 7 8 9 10 Genome size (Mb) 3.5 5.2 4.8 4.4 4.3 4.4 2.9 3.2 2.9 3.6 GC content (%) 51.4 64.5 64.5 60.5 62.2 63.4 63.5 59.5 68.3 64.5 Number of genes 3408 4839 4529 4198 4125 4064 2805 3153 2845 3454 Number of CDS 3274 4733 4407 4099 4010 3979 2717 3041 2760 3333 Phenotypic and Physiological Characteristics Colonies of strain TC5R-5 T were white and non-transparent. The cells were rod-shaped, measuring approximately 0.8–0.9 µm in width and 3.0–4.0 µm in length, and exhibited a single polar flagellum (Fig. 3 ). Cells were Gram-stain-negative, catalase- and oxidase-positive, but did not display DNase activity. Growth occurred only under aerobic conditions. The strain tolerated NaCl concentrations ranging from 0 to 0.5% (w/v), and was capable of growth at temperatures between 15–30°C and pH 6.0–8.0, with optimal growth observed at 20–30°C and pH 6.0–7.0. Substrate degradation assays were performed on R2A agar supplemented with various compounds (5.0% casein, 1.0% aesculin, 1.0% Tween 20, 1.0% Tween 80, 1.0% starch; w/v). Strain TC5R-5 T demonstrated limited carbon source utilization and acid production capabilities. It also tested negative for numerous enzymatic activities and biochemical reactions when compared to type strains of related genera within the family Chromobacteriaceae . A summary of its morphological and physiological characteristics is provided in Table 2 . Table 2 Differential characteristics between strain TC5R-5 T and most related phylogenetic neighbors Strains: 1. TC5R-5 T ; 2. Chromobacterium alkanivorans IITR-71 T ; 3. Chromobacterium violaceum ATCC 12472 T ; 4. Aquitalea magnusonii TRO-001DR8 T ; 5. Paludibacterium yongneupense 5YN8-15 T ; 6. Pseudogulbenkiania subflava DSM 22618 T ; 7. Gulbenkiania mobilis E4FC31 T ; 8. Craterilacusibacter sinensis B2N2-7 T ; 9. Crenobacter luteus YIM 78141 T ; 10. Vogesella indigofera DSM 3303 T . +, Positive; -, negative; w, weakly positive. All of the data are acquired from this experiment except where indicated otherwise. a,b,c,d,e,f Data from corresponding literatures [Liu et al., 2020 ; Dong et al., 2015 ; Subhash et al., 2013 ; Sedláček et al., 2016 ; Sheu et al., 2014 ; Vaz-Moreira et al., 2007 ]. 1 2 3 4 5 6 7 8 a, 9 b 10 c Reactions (API 20E&20NE) Fermentation of glucose - + + + + - - - + - Arginine dihydrolase - + + + - - + + - + Hydrolysis of gelatin - + + - - - - - - - Enzyme activity (API ZYM ) α -Chymotrypsin - - + - + + - - - - N-Acetyl- β - glucosaminidase - + + - w - - - - - Utilization of substrates (Biolog GENIII) D-Trehalose - w w - + + - - + - Inosine - + w - - - - - - - Glycerol - + w + - - - - - + Gelatin - + + - - - - - - - L-Arginine - w w + - + + w + + Pectin w - - - + + - w + - Mucic acid w - - - w - - - - - α –Keto-glutaric acid + w - + w - - w - - Formic acid + - w w - + - w - + Acids production (API 50CH) D-Glucose + - + + + + w - + w D- Mannose - - + - - - - - - - Trehalose - + + - + + - - - - Respiratory quinone Q-8, Q-7 Q-8, Q-7 ND Q-8 d Q-8 e Q-8 e Q-8, Q-9 f Q-8 Q-8 ND Polar Lipid PE + + ND + d + [e] ND ND + + + c PG + + ND + + ND ND + - + DPG + + ND + + ND ND + + + APL 1 1 ND 1 1 ND ND 2 - 3 AL - 4 ND - - ND ND - - - PL 1 - ND 2 5 ND ND - 3 1 L - - ND 2 - ND ND 3 - - AGP - - ND - - ND ND - 1 - Chemotaxonomic Characteristics The cellular fatty acid profile of strain TC5R-5 T was presented in Table 3 . The predominant component was summed feature 3 (C 16:1 ω 7 c /C 16:1 ω 6 c ; 51.6%), which was notably higher than those observed in related reference strains. In contrast, the content of summed feature 8 (C 18:1 ω 7c/C 18:1 ω 6c) in strain TC5R-5 T was less than half of that detected in Chromobacterium alkanivorans IITR-71 T . Overall, the principal fatty acids in strain TC5R-5 T and all type strains of the family Chromobacteriaceae were summed were summed feature 3 (C 16:1 ω 7 c /C 16:1 ω 6 c ) and C 16:0 . The major polar lipids of strain TC5R-5 T were phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), one unidentified aminophospholipid (APL1), and one unidentified phospholipid (PL1) (Figure S4). For comparative classification, the polar lipid profile of Chromobacterium alkanivorans IITR-71 T was also examined. The results indicated the presence of four aminolipids and one aminophospholipid in Chromobacterium alkanivorans IITR-71 T , which were absent in TC5R-5 T . Moreover, the comparison between strain TC5R-5 T and the type species of related genera in the family Chromobacteriaceae were also conducted. As shown in Table 2 , polar lipids of most of type strains, except Crenobacter luteus YIM 78141 T , consisted of PE, PG and DPG. The predominant respiratory quinone of strain TC5R-5 T was ubiquinone 8 (Q-8), being in line with the quinone profile of members of the family Chromobacteriaceae . Table 3 Whole-cell fatty acid profiles of the novel isolates and the type strains of related species of genera 1.strain TC5R-5 T ; 2. Chromobacterium alkanivorans IITR-71 T ; 3. Chromobacterium violaceum ATCC 12472 T ; 4. Aquitalea magnusonii TRO-001DR8 T ; 5. Paludibacterium yongneupense 5YN8-15 T ; 6. Pseudogulbenkiania subflava DSM 22618 T ; 7. Gulbenkiania mobilis E4FC31 T ; 8. Craterilacusibacter sinensis B2N2-7 T ; 9. Crenobacter luteus YIM 78141 T ; 10. Vogesella indigofera DSM 3303 T . All data were from this study unless otherwise indicated. *Summed features are groups of two acids that cannot be separated by GLC using the MIDI system. a Data from Lau et al., b Data from Liu et al., c Data from Dong et al. 1 2 3 a 4 5 6 7 8 b 9 c 10 b C 12:0 7.1 10.5 5.6 7.2 2.9 6.5 3.6 2.0 5.1 3.8 C 14:0 2.2 3.1 1.7 1.3 3.8 0.8 0.8 3.6 - 0.5 C 16:0 26.7 20.5 23.9 30.5 29.8 28.7 24.9 33.8 27.2 24.9 C 10:0 3-OH 3.6 4.6 5.2 3.6 - 3.3 3.2 2.9 2.6 7.2 C 12:0 3-OH 2.7 2.2 5.0 2.2 3.5 2.6 3.2 3.3 2.3 - Summed feature 3* 51.6 44.8 35.8 40.8 34.4 38.2 43.5 31.1 44.0 45.6 Summed feature 8* 5.1 12.1 15.0 10.7 13.2 11.8 9.1 1.0 - 7.9 * Summed Features are fatty acids that cannot be resolved reliably from another fatty acid using the chromatographic conditions chosen. The MIDI system groups these fatty acids together as one feature with a single percentage of the total. Summed feature 3, C 16:1 ω 7 c /C 16:1 ω 6 c ; Summed feature 8, C 18:1 ω 7 c /C 18:1 ω 6 c . Data from corresponding literatures. Taxonomic Conclusion We conducted comprehensive phylogenetic analysis based on the 16S rRNA gene and whole genome alignments to determine the phylogenetic position of strain TC5R-5 T . Phylogenetic trees reconstructed from the 16S rRNA gene consistently placed the strain within the family Chromobacteriaceae . This assignment was further supported by phylogenomic analyses performed with IQ-TREE using single-copy genes and with UBGC based on core genes. In contrast, the phylogenomic tree generated with the TYGS online tool showed that strain TC5R-5ᵀ formed a cluster with Halomonas humidisoli WN018ᵀ, Vreelandella glaciei DD39ᵀ, and Andreprevotia chitinilytica JS11-7ᵀ. Subsequently, we calculated the Average Nucleotide Identity (ANI) and genome coverage between strain TC5R-5 T with its phylogenetic neighbors identified in the phylogenomic trees (Table S1 ). The results showed that strain TC5R-5 T shared higher ANI values with Chromobacterium alkanivorans IITR-71 T and Aquitalea palustris CCM7557 T (73.48%, 72.49%) than with Andreprevotia chitinilytica DSM 18519 T , Halomonas humidisoli WN018 T , and Vreelandella glaciei DD39 T (68.49%, 64.26%, 64.73%). Genome coverage was also notably greater with the former group (26.78% and 25.03%, respectively) compared to the latter (9.92%, 5.01%, and 4.91%, respectively). These findings further corrobrorated that strain TC5R-5ᵀ is phylogenetically affiliated with the family Chromobacteriaceae . In summary, comprehensive phylogenetic analyses based on the 16S rRNA gene and whole-genome sequences consistently support the classification of strain TC5R-5 T as representing a novel genus and species within the order Neisseriales . Although minor topological variations were observed among phylogenomic trees constructed using different analytical methods, the overall evidence strongly supported its provisional classification within the family Chromobacteriaceae as the most coherent and biologically plausible placement. Description of Craterilacuimicrobium gen. nov. Craterilacuimicrobium gen. nov. (Cra.te.ri.la.cu.i.mi.cro'bium. L. masc. n. crater , a crater; L. masc. n. lacus , a lake; N.L. neut. n. microbium , a microbe; N.L. neut. n. Craterilacuimicrobium , a microbe from a crater lake) Cells rod-shaped, motile, Gram-stain-negative and strictly aerobic. Catalase- and oxidase-positive. The major fatty acids consist of summed feature 3 (C 16:1 ω 7 c /C 16:1 ω 6 c ). The predominant ubiquinone is Q-8, with a small amount of Q-7. The main polar lipids include phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), one unidentified aminophospholipid (APL), and one unidentified phospholipid (PL). The type species is Craterilacuimicrobium aquaticum . Description of Craterilacuimicrobium aquaticum sp. nov. Craterilacuimicrobium aquaticum sp. nov. (a.qua'ti.cum. L. neut. adj. aquaticum , living, growing or found in or by water, aquatic) Cells are Gram-stain-negative, strictly aerobic, motile, and short rod-shaped. Colonies are circular, smooth, white-colored after incubating for 48 h at 25°C on R2A agar. Growth occurs at 15–30°C (optimum, 20–30°C), at pH 6.0–8.0 (optimum, 6.0–7.0) and with 0-0.5% NaCl (optimum, 0-0.5%, w/v). Catalase- and oxidase-activity present. DNase activity is not present. Positive for hydrolyzation of Tween 20, but negative for degradation of casein, cellulose, starch, aesculin and Tween 80. In the API 20 NE and 20E tests, strain TC5R-5 T could reduce nitrate to nitrite. Voges-Proskauer-test and fermentation of glucose are positive. Citrate not utilized, indole not produced, no arginine dihydrolase activity, no hydrolysis of gelatin and esculin. Does not produce H 2 S. The activities of ornithine decarboxylase, lysine decarboxylase, tryptophane deaminase, urease, ortho-nitrophenyl- β -D-galactopyranosidase and para-nitrophenyl- β -D-galactopyranosidase are negative. In the API ZYM test, positive for alkaline phosphatase, leucine arylamidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase, but negative for esterase (C4), esterase lipase (C8), lipase (C14), valine arylamidase, cystine arylamidase, trypsin, α -chymotrypsin, α -galactosidase, β -galactosidase, β -glucuronidase, α -glucosidase, β -glucosidase, N -acetyl- β -glucosaminidase, α -mannosidase and α -fucosidase. Acids could be produced from glucose in API 50CH, but not other carbon sources. In the BIOLOG GEN III MicroPlate system, α -D-glucose, D-glucose-6-PO 4 , L-alanin, L-glutamic acid, L-histidine, L-serine, methyl pyruvate, L-lactic acid, α -keto-glutaric acid, γ -amino-butrytic acid, β –hydroxyl-D, L-butyric acid, acetoacetic acid, formic acid are utilized as sole carbon resource, and dextrin, D-frucose, D-fucose, L-fucose, D-fructose-6-PO 4 , D-serine, glycyl-L-proline, L-aspartic acid, L-pyroglutamic acid, pectin, D-galacturonic acid, D-glucuronic acid, glucuronamide, mucic acid, D-lactic acid metyl ester, citric acid, D-malic acid, L-malic acid, Tween 40, α -hydroxy-butyric acid, propionic acid, acetic acid are weakly utilized. The genomic DNA G + C content is 51.4% according to the genomic sequencing data. The major fatty acids (> 10%) are summed feature 3 (C 16:1 ω 7 c /C 16:1 ω 6 c ) and C 16:0 . The predominant respiratory quinone is Q-8 with a trace amount of Q-7. The main polar lipids are phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), one unidentified aminophospholipid (APL) and one unidentified phospholipid (PL). The type strain TC5R-5 T (= CGMCC 1.17058 T = KCTC 72736 T ) was isolated from water of Aershan Tianchi crater lake in the Inner Mongolia Autonomous Region of China. The GenBank/EMBL/DDBJ accession number for 16S rRNA gene of strain TC5R-5 T is OK487579 and the DDBJ/ENA/GenBank accession number of the Whole Genome Shotgun project of TC5R-5 T is JACCFC000000000. Abbreviations ANI, average nucleotide identity; APL, unidentified aminophospholipid; CDS, coding sequences; DPG, diphosphatidylglycerol; PE, phosphatidylethanolamine; PG, phosphatidylglycerol; PL, unidentified phospholipid; AGP, aminoglycophospholipid; L, unidentified polar lipid; NRPS, nonribosomal peptide synthetase. Declarations Conflicts of interest: The authors declare that there are no conflicts of interest. Ethical approval: The data reported in this article is not derived from any studies or tests on human volunteers or animals. Informed consent: All the authors have read and approved the final manuscript for submission. Funding information This work was financially supported by the National Natural Science Foundation of China (31970004), the Strategic Biological Resources Capacity Building Project of Chinese Academy of Sciences, China (KFJ-BRP-017-73) and the Key Research and Development Project of Xizang Autonomous Region, China (XZ202401ZY0061). Author Contribution You-Jun Liao: Conceptualization, Resources, Data curation, Methodology; Xuan Zhang : Investigation, Formal analysis; Zi-Xuan Liu: Visualization, Writing—Original Draft and Editing; Ai-Hua Li: Funding acquisition, Project administration, Writing—Reviewing. Acknowledgement The authors gratefully acknowledge the help from Prof. Teun Boekhout (College of Sciences, King Saud University, Saudi Arabia) to improve the English text, the help from Dr. Jing-Nan Liang for the TEM observation, and the assistance from Prof. Stefano Ventura (Research institute on Terrestrial Ecosystems, Firenze Unit, Italy) for his advice on bacterial nomenculature. Data Availability The GenBank/EMBL/DDBJ accession number for 16S rRNA gene of strain TC5R-5T is OK487579. The Whole Genome Shotgun project of strain TC5R-5T has been deposited at DDBJ/ENA/GenBank under the accession number JACCFC000000000. This strain is available at China General Microbiology Culture Collection Center (CGMCC) under the accession no. CGMCC 1.17058T. References Adeolu M, Gupta RS (2013) Phylogenomics and molecular signatures for the order Neisseriales : proposal for division of the order Neisseriales into the emended family Neisseriaceae and Chromobacteriaceae fam. nov. Antonie Van Leeuwenhoek 104:1–24. https://doi.org/10.1007/s10482-013-9920-6 . Aziz RK, Bartels D, Best AA et al (2008) The rast server: rapid annotations using subsystems technology. BMC Genomics 9:75–75. https://doi.org/10.1186/1471-2164-9-75 . Bajaj A, Kumar A, Yadav S, Kaur G, Bala M et al. (2016) Isolation and characterization of a novel Gram-negative bacterium Chromobacterium alkanivorans sp. nov., strain IITR-71 T degrading halogenated alkanes. Int J Syst Evol Microbiol 66:5228–5235. https://doi.org/10.1099/ijsem.0.001500 . Chen S, Rudra B, Gupta RS (2021) Phylogenomics and molecular signatures support division of the order Neisseriales into emended families Neisseriaceae and Chromobacteriaceae and three new families Aquaspirillaceae fam. nov., Chitinibacteraceae fam. nov., and Leeiaceae fam. nov. Syst Appl Microbiol 44:126251. https://doi.org/10.1016/j.syapm.2021.126251 Chun J, Oren A, Ventosa A et al (2018) Proposed minimal standards for the use of genome data for the taxonomy of prokaryotes. Int J Syst Evol Microbiol 68:461–466. https://doi.org/10.1099/ijsem.0.002516 Chou JH, Chou YJ, Arun AB, et al (2009) Vogesella lacus sp. nov., isolated from a soft-shell turtle culture pond. Int J Syst Evol Microbiol 59:2629–2632. https://doi.org/10.1099/ijs.0.009266-0 Collins MD (1985) Isoprenoid quinone analysis in classification and identification. In: Goodfellow M and Minnikin DE (ed). Chemical Methods in Bacterial Systematics . Academic Press, London, pp 267–287. De Ley J, Segers P, Gillis M. (1978) Intra- and Intergeneric Similarities of Chromobacterium and Janthinobacterium Ribosomal Ribonucleic Acid Cistrons. Int. J. Syst. Bacteriol. 1978; 28:154–168. https://doi.org/10.1099/00207713-28-2-154 . Dong L, Ming H, Zhou EM, et al (2015) Crenobacter luteus gen. nov., sp. nov., isolated from a hot spring. Int J Syst Evol Microbiol 65:214–219. https://doi.org/10.1099/ijs.0.060996-0 Felsenstein J (1981) Evolutionary trees from DNA sequences: a maximum likelihood approach. J Mol Evol 17:368–376. https://doi.org/10.1007/BF01734359 Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791. https://doi.org/10.1111/j.1558-5646.1985.tb00420.x Fitch WM (1971) Toward defining the course of evolution: Minimum change for a specific tree topology. Syst Zool 20:406–416. https://doi.org/10.1093/sysbio/20.4.406 . Gerhardt P, Murray RGE, Wood WA, Krieg NR (1994) Methods for General and Molecular Bacteriology. Amer Soc Microbiol.Washington DC. Grimes DJ, Woese CR, MacDonell MT, Colwell RR (1997) Systematic study of the genus Vogesella gen. nov. and its type species, Vogesella indigofera comb. nov. Int J Syst Bacteriol 47:19–27. https://doi.org/10.1099/00207713-47-1-19 Jyoti V, Narayan KD, Das SK (2010) Gulbenkiania indica sp. nov., isolated from a sulfur spring. Int J Syst Evol Microbiol 60:1052–1055. https://doi.org/10.1099/ijs.0.014035-0 Kai B, Shaw S, Kloosterman AM, Charlop-Powers Z, Weber T (2021) Antismash 6.0: improving cluster detection and comparison capabilities. Nucleic Acids Res. 49(W1): W29-W35. https://doi.org/10.1093/nar/gkab335 Kimura M (1980) A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16:111–120. https://doi.org/10.1007/BF01731581 Komagata K, Suzuki K (1987) Lipid and cell-wall analysis in bacterial systematic. Methods Microbiol 19:161–207. https://doi.org/10.1016/S0580-9517(08)70410-0 Kwon SW, Kim BY, Kim WG, et al (2008) Paludibacterium yongneupense gen. nov., sp. nov., isolated from a wetland, Yongneup, in Korea. Int J Syst Evol Microbiol 58:190–194. https://doi.org/10.1099/ijs.0.64831-0 Lane DJ, Goodfellow M (1991) 16S/23S rRNA sequencing. In: E. Stackebrandt. Chichester (ed). Nucleic Acid Techniques in Bacterial Systematics, Wiley, New York, pp 115–175. Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA et al (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23:2947–2948. https://doi.org/10.1093/bioinformatics/btm404 Lau HT, Faryna J, Triplett EW (2006) Aquitalea magnusonii gen. nov., sp. nov., a novel Gram-negative bacterium isolated from a humic lake. Int J Syst Evol Microbiol 56:867–871. https://doi.org/10.1099/ijs.0.64089-0 Lin MC, Chou JH, Arun AB, Young CC, Chen WM (2008) Pseudogulbenkiania subflava gen. nov., sp. nov., isolated from a cold spring. Int J Syst Evol Microbiol 58:2384–2388. https://doi.org/10.1099/ijs.0.65755-0 Liu ZX, Phurbu D, Liu HC, Zhou YG, Li AH (2020) Craterilacuibacter sinensis gen. nov. sp. nov., isolated from a crater lake in China. Int J Syst Evol Microbiol 70:4831–4837. https://doi.org/10.1099/ijsem.0.004353 Luo C, Rodriguez-R LM, Konstantinidis KT (2014) MyTaxa: an advanced taxonomic classifier for genomic and metagenomic sequences. Nucleic Acids Res 42:e73. https://doi.org/10.1093/nar/gku169 Meier-Kolthoff JP, Göker M. TYGS is an automated high-throughput platform for state-of-the-art genome-based taxonomy. Nat Commun. 2019;10:2182. https://doi.org/10.1038/s41467-019-10210-3 Miller LT (1982) Single derivatization method for routine analysis of bacterial whole-cell fatty acid methyl esters, including hydroxy acids. J Clin Microbiol 16:584–586. https://doi.org/10.1128/jcm.16.3.584-586.1982 Minh BQ, Schmidt HA, Chernomor O, et al (2020) IQ-TREE 2: New Models and Efficient Methods for Phylogenetic Inference in the Genomic Era. Mol Biol Evol. 37:2461. https://doi.org/10.1093/molbev/msaa131 Minnikin DE, Bolton RC, Hartmann S et al. (1993) An integrated procedure for the direct detection of characteristic lipids in tuberculosis patients. Ann Soc Belg Med Trop 73 Suppl 1:13–24. Richter M, Rosselló-Móra R. (2009) Shifting the genomic gold standard for the prokaryotic species definition. Proc Natl Acad Sci USA 106:19126–19131. https://doi.org/10.1073/pnas.0906412106 Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425. https://doi.org/10.1093/oxfordjournals.molbev.a040454 Sasser M (1990) Identification of bacteria by gas chromatography of cellular fatty acids, MIDI Technical Note 101. NewYork: MIDI Inc Sedláček I, Kwon SW, Švec P, Mašlanŏvá I, Kýrová K, et al (2016) Aquitalea pelogenes sp. nov., isolated from mineral peloid. Int J Syst Evol Microbiol. 66:962–967. https://doi.org/10.1099/ijsem.0.000819 Sheu SY, Chen ZH, Young CC, Chen WM (2014) Paludibacterium paludis sp. nov., isolated from a marsh. Int J Syst Evol Microbiol 64:2497–2502. https://doi.org/10.1099/ijs.0.060954-0 Subhash Y, Tushar L, Sasikala C, Ramana CV (2013) Vogesella alkaliphila sp. nov., isolated from an alkaline soil, and emended description of the genus Vogesella . Int J Syst Evol Microbiol. 63:2338–2343. https://doi.org/10.1099/ijs.0.046300-0 Tamura K, Peterson D, Peterson N, Stecher G, Nei M et al (2011) MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. Molecular Biology and Evolution 10:2731–2739. https://doi.org/10.1093/molbev/msr121 Vaz-Moreira I, Nobre MF, Nunes OC, Manaia CM (2007) Gulbenkiania mobilis gen. nov., sp. nov., isolated from treated municipal wastewater. Int J Syst Evol Microbiol 57:1108–1112. https://doi.org/10.1099/ijs.0.64726-0 Winey M, Meehl JB, O'Toole ET, Giddings TH Jr. (2014) Conventional transmission electron microscopy. Mol Biol Cell 25:319–23. https://doi.org/10.1091/mbc.E12-12-0863 Wu C, Lu X, Qin M et al (1989) Analysis of menaquinone compound in microbial cells by HPLC. Microbiology [English translation of Microbiology (Beijing)] 16:176–178. Yoon SH, Ha SM, Kwon S et al. (2017) Introducing EzBioCloud: a taxonomically united database of 16S rRNA gene sequences and whole-genome assemblies. Int J Syst Evol Microbiol 67:1613–1617. https://doi.org/10.1099/ijsem.0.001755 . Additional Declarations No competing interests reported. Supplementary Files supplementaryofTC5R5T2025.9.12.pdf Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 05 Feb, 2026 Reviews received at journal 26 Jan, 2026 Reviewers agreed at journal 20 Nov, 2025 Reviews received at journal 13 Oct, 2025 Reviewers agreed at journal 07 Oct, 2025 Reviewers invited by journal 06 Oct, 2025 Editor assigned by journal 12 Sep, 2025 Submission checks completed at journal 12 Sep, 2025 First submitted to journal 11 Sep, 2025 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. We do this by developing innovative software and high quality services for the global research community. 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06:22:12","extension":"html","order_by":15,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":160707,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7595832/v1/ed5f936f54450db8d3b86729.html"},{"id":93825371,"identity":"b6e51567-efe3-4c25-be36-f6922ef81102","added_by":"auto","created_at":"2025-10-18 06:14:11","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":90857,"visible":true,"origin":"","legend":"\u003cp\u003eMaximum-likelihood phylogenetic tree based on 16S rRNA gene sequences of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e. Genbank accession numbers are shown in parentheses. Bootstrap values are expressed as percentages of 1000 replications and shown at branching nodes (\u0026gt;50%). The filled circles indicate nodes recovered using the Neighbor-joining and Maximum-parsimony methods. Bar 0.02 substitution per nucleotide position.\u003c/p\u003e","description":"","filename":"Picture1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7595832/v1/53095401710c57c0a2b5c5c5.jpg"},{"id":93825372,"identity":"940d62dd-cff6-486b-9964-b1ce5929fbd7","added_by":"auto","created_at":"2025-10-18 06:14:11","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":328711,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic trees based on the genome sequences showing the phylogenetic position of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e in the within the order\u003cem\u003e Neisseriales.\u003c/em\u003e (a) the phylogenomic tree reconstructed by IQ-TREE based on single copy gene with MFP as model used and 1000 ultrafast bootstrap repeats; Bootstrap values of 100 have been removed from the nodes; (b) the phylogenomic tree based on core genes drawn by using the UBCG 2.0; (c) the phylogenomic tree obtained using the TYGS platform. Branch lengths are scaled in GBDP distance. The numbers above branches were GBDP pseudo-bootstrap support values of \u0026gt;50% from 1000 replications.\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7595832/v1/cb7ea43519cd90da91243697.jpg"},{"id":93825373,"identity":"3a7b17ea-bdc6-48cc-9a52-1d017d2682f1","added_by":"auto","created_at":"2025-10-18 06:14:11","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":76136,"visible":true,"origin":"","legend":"\u003cp\u003eTransmission electron micrograph of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e after being cultivated on R2A agar at 25 ℃ for 48h.\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7595832/v1/9aed088a2e2dc4e987212e46.jpg"},{"id":93826170,"identity":"837f7b65-3f06-4265-9bf9-815c25c7c6d6","added_by":"auto","created_at":"2025-10-18 06:38:12","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1774505,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7595832/v1/ab33e2f5-04b1-4749-831e-ec73393564b2.pdf"},{"id":93825376,"identity":"b47ae954-92fe-417d-9bd5-dce9c298bdb7","added_by":"auto","created_at":"2025-10-18 06:14:11","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":735638,"visible":true,"origin":"","legend":"","description":"","filename":"supplementaryofTC5R5T2025.9.12.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7595832/v1/d919bc00d3c044454c06511b.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Craterilacuimicrobium aquaticum gen. nov. sp. nov., a novel member of the order Neisseriales isolated from crater lake","fulltext":[{"header":"Introduction","content":"\u003cp\u003eThe order \u003cem\u003eNeisseriales\u003c/em\u003e was reorganized by Adeolu \u003cem\u003eet al\u003c/em\u003e. in 2013, dividing it into the families \u003cem\u003eNeisseriaceae\u003c/em\u003e and \u003cem\u003eChromobacteriaceae\u003c/em\u003e based on phylogenomic and molecular signatures [Adeolu and Gupta, \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2013\u003c/span\u003e]. Chen \u003cem\u003eet al.\u003c/em\u003e classified 37 genera within the order \u003cem\u003eNeisseriales\u003c/em\u003e, encompassing 122 species, into two revised families \u003cem\u003eNeisseriaceae\u003c/em\u003e and \u003cem\u003eChromobacteriaceae\u003c/em\u003e, along with three new families namely \u003cem\u003eAquaspirillaceae\u003c/em\u003e fam. nov., \u003cem\u003eChitinibacteraceae\u003c/em\u003e fam. nov., and \u003cem\u003eLeeiaceae\u003c/em\u003e fam. nov. [Chen et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2021\u003c/span\u003e]. The family \u003cem\u003eChromobacteriaceae\u003c/em\u003e contained the type genus \u003cem\u003eChromobacterium\u003c/em\u003e [De Ley et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1978\u003c/span\u003e] and other genera like \u003cem\u003eAquitalea\u003c/em\u003e [Lau et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2006\u003c/span\u003e], \u003cem\u003eCraterilacuibacter\u003c/em\u003e [Liu et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2020\u003c/span\u003e], \u003cem\u003eCrenobacter\u003c/em\u003e [Dong et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2015\u003c/span\u003e], \u003cem\u003eGulbenkiania\u003c/em\u003e [Vaz-Moreira et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2007\u003c/span\u003e], \u003cem\u003ePaludibacterium\u003c/em\u003e [Kwon et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2008\u003c/span\u003e], \u003cem\u003ePseudogulbenkiania\u003c/em\u003e [Lin et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2008\u003c/span\u003e] and \u003cem\u003eVogesella\u003c/em\u003e [Grimes et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e1997\u003c/span\u003e]. Strains of these genera were predominantly isolated from aqueous and soil habitats, such as soil, rice field and water samples. \u003cem\u003eAquitalea magnusonii\u003c/em\u003e [Lau et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2006\u003c/span\u003e], \u003cem\u003eGulbenkiania indica\u003c/em\u003e [Jyoti et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2010\u003c/span\u003e], \u003cem\u003eGulbenkiania mobilis\u003c/em\u003e [Vaz-Moreira et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2007\u003c/span\u003e], \u003cem\u003eVogesella lacus\u003c/em\u003e [Chou et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2009\u003c/span\u003e], and \u003cem\u003ePseudogulbenkiania subflava\u003c/em\u003e [Lin et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2008\u003c/span\u003e] were isolated from various aquatic environments, such as humic lake, sulfur spring, cold spring and pond.\u003c/p\u003e\u003cp\u003eDuring the exploration of bacterial diversity of the Aershan Tianchi crater lake, strain TC5R-5\u003csup\u003eT\u003c/sup\u003e was isolated from a water sample. Due to its low 16S rRNA similarity (94.3%) with the currently valid published species, a series of experiments were conducted to determine the taxonomic status of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e. Using polyphasic taxonomy, strain TC5R-5\u003csup\u003eT\u003c/sup\u003e was classified as a novel species of a new genus within the order \u003cem\u003eNeisseriales.\u003c/em\u003e Despite minor topological discrepancies among phylogenomic trees generated through different analytical methods, the collective evidence strongly supports its provisional assignment to the family \u003cem\u003eChromobacteriaceae\u003c/em\u003e as the most coherent and biologically plausible placement.\u003c/p\u003e"},{"header":"Material and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eStrain Isolation\u003c/h2\u003e\u003cp\u003eStrain TC5R-5\u003csup\u003eT\u003c/sup\u003e was isolated from water samples collected from the surface water at a depth of 0.5 m in Aershan Tianchi crater lake, September 2015, located on Da Hinggan Ling Mountain in northern China (120.41\u0026deg;E, 47.32\u0026deg;N). The water temperature was 12\u0026deg;C. The samples were diluted from 10\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e to 10\u003csup\u003e\u0026minus;\u0026thinsp;5\u003c/sup\u003e gradient with sterile 0.85% NaCl (w/v) and cultured on R2A agar medium (Difco) at 25\u0026deg;C for 7 days. Single colonies were picked up and purified by streaking on R2A plate once again. The strains were preserved by lyophilization and in liquid nitrogen, respectively.\u003c/p\u003e\u003cp\u003e\u003cb\u003e16S rRNA Gene Sequencing and Phylogenetic Analysis\u003c/b\u003e\u003c/p\u003e\u003cp\u003eAfter being cultivated on R2A agar at 25℃, the genomic DNA of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e was extracted using the Genomic DNA Rapid Isolation Kit for bacterial cells (BioDev-Tech. Beijing, China) and the 16S rRNA gene of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e was amplified with the universal primers 27F and 1492R [Lane and Goodfellow, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e1991\u003c/span\u003e]. An almost complete 16S rRNA gene sequence were obtained and analysed using the GenBank databases (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://blast.ncbi.nlm.nih.gov/Blast.cgi\u003c/span\u003e\u003cspan address=\"https://blast.ncbi.nlm.nih.gov/Blast.cgi\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) and EzBioCloud (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ezbiocloud.net/\u003c/span\u003e\u003cspan address=\"https://www.ezbiocloud.net/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) [Yoon et al., \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2017\u003c/span\u003e]. Sequences of related taxa were downloaded and aligned using Clustal W [Larkin et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2007\u003c/span\u003e]. Phylogenetic trees were constructed using neighbor-joining, maximum-likelihood and maximum-parsimony methods in MEGA7.0 [Felsenstein, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e1981\u003c/span\u003e; Fitch, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e1971\u003c/span\u003e; Saitou and Nei, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e1987\u003c/span\u003e; Tamura et al., \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2011\u003c/span\u003e]. The evolutionary distance matrices were calculated using Kimura\u0026rsquo;s two parameter model [Kimura, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e1980\u003c/span\u003e], and bootstrap values were determined based on 1000 replications [Felsenstein, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e1985\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eGenome Features and Phylogenomic Analysis\u003c/h3\u003e\n\u003cp\u003eThe genome of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e was sequenced using the Illumina HiSeq X platform at Majorbio Co., Ltd. The qualified samples were pair-end sequenced, and raw sequencing data were acquired. Quality control used SeqPrep v0.20.0, the genome sequence was de novo assembled using SOAP v2.04. Contigs exceeding 400 bp in length were retained for gene prediction. CDS prediction was carried out using Glimmer v3.02 (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://ccb.jhu.edu/software/glimmer/index.shtml\u003c/span\u003e\u003cspan address=\"http://ccb.jhu.edu/software/glimmer/index.shtml\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) and GeneMarkS v4.3 (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://topaz.gatech.edu/GeneMark\u003c/span\u003e\u003cspan address=\"http://topaz.gatech.edu/GeneMark\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e). The rRNA and tRNA were predicted using Barrnap v0.8; (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://github.com/tseemann/barrnap/\u003c/span\u003e\u003cspan address=\"https://github.com/tseemann/barrnap/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) and tRNA-scan-SE v2.0 (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://trna.ucsc.edu/software/\u003c/span\u003e\u003cspan address=\"http://trna.ucsc.edu/software/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e), respectively. Average Nucleotide Identity (ANI) was determined through the OrthoANI algorithm on Ezbiocloud [Yoon et al., \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2017\u003c/span\u003e]. The dDDH values were calculated using the TYGS with the GBDP method [Meier-Kolthoff and G\u0026ouml;ker, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2019\u003c/span\u003e]. Amino acid identity (AAI) was calculated using an AAI calculator (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://enve-omics.ce\u003c/span\u003e\u003cspan address=\"http://enve-omics.ce\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. gatech.edu/aai/index) [Luo et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2014\u003c/span\u003e]. The ANI, dDDH and AAI values were compared with the thresholds established in literature [Chun et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Richter and Rossell\u0026oacute;-M\u0026oacute;ra, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2009\u003c/span\u003e]. The genes were classified into subsystems and clusters of orthologous groups (COGs) using the Rapid Annotation using Subsystem Technology (RAST) server [Aziz et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2008\u003c/span\u003e]. Genomic annotation was carried out using the NR (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003eftp://ftp.ncbi.nlm.nih.gov/blast/db/\u003c/span\u003e\u003cspan address=\"http://ftp://ftp.ncbi.nlm.nih.gov/blast/db/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e), KEGG (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://www.genome.jp/kegg/\u003c/span\u003e\u003cspan address=\"http://www.genome.jp/kegg/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) and eggNOG (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://eggnogdb.embl.de/#/app/home\u003c/span\u003e\u003cspan address=\"http://eggnogdb.embl.de/#/app/home\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) algorithms, respectively. The software antiSMASH 6.0 was utilized to predict the genes responsible for the secondary metabolites [Kai et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2021\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eTo confirmed the phylogenetic position of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e, three genome-based phylogenetic trees were reconstructed using different methods: one used the TYGS platform [Meier-Kolthoff and G\u0026ouml;ker, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2019\u003c/span\u003e], another was reconstructed with IQ-TREE v2.1.2 and based on single copy genes using MFP as the model and 1000 ultrafast bootstrap repeats [Minh et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2020\u003c/span\u003e], and the third phylogenomic tree was generated based on core genes using UBGC2.0 on EzBiocloud.\u003c/p\u003e\n\u003ch3\u003eMorphological and Physiological Analyses\u003c/h3\u003e\n\u003cp\u003eThe morphological and physiological characteristics were determined through the following experiments. Cell morphology was observed using phase-contrast microscope and transmission electron microscope (JEM1400; JEOL) after 2 days of growth on R2A agar at 25℃ [Winey et al., \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2014\u003c/span\u003e]. Cells were stained with uranium acetate before observation. Motility was assessed by inoculating strain TC5R-5\u003csup\u003eT\u003c/sup\u003e into semi-solid R2A agar medium (0.5% agar) and observing its growth trace in the medium. A single colony of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e was circular, smooth, and white-colored. Cells were 0.8\u0026ndash;0.9 \u0026micro;m in width and 3.0\u0026ndash;4.0 \u0026micro;m in length. The optimal growth conditions of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e, including temperature range, pH and NaCl concentrations were confirmed as described below. Strain TC5R-5\u003csup\u003eT\u003c/sup\u003e was cultured in R2A broth with NaCl concentrations ranging from 0 to 5.0% (w/v, at intervals of 0.5%). The pH variations (4.0, 5.0, 6.0, 6.5, 7.0, 7.5, 8.0, 9.0, 10.0) was achieved using specific buffers: 100mM acetate buffer (for pH 4.0\u0026ndash;5.0), 100mM phosphate buffer (pH 6.0\u0026ndash;8.0) or 100mM NaHCO\u003csub\u003e3\u003c/sub\u003e/Na\u003csub\u003e2\u003c/sub\u003eCO\u003csub\u003e3\u003c/sub\u003e buffer (pH 9.0\u0026ndash;10.0). Growth at different temperatures (4, 15, 20, 25, 30, 35, 37, 40 and 45\u0026deg;C) was evaluated in R2A broth under the optimal NaCl concentration. Gram-staining, anaerobic growth and DNase activity were measured following previously described methods [Gerhardt et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1994\u003c/span\u003e]. The catalase and oxidase activities were assessed using 3% H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e solution and 1% tetra-methyl-\u003cem\u003ep\u003c/em\u003e-phenylenediamine (bioM\u0026eacute;rieux), respectively. Strain TC5R-5\u003csup\u003eT\u003c/sup\u003e and its related reference strains, such as \u003cem\u003eChromobacterium alkanivorans\u003c/em\u003e IITR-71\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eChromobacterium violaceum\u003c/em\u003e ATCC 12472\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eAquitalea magnusonii\u003c/em\u003e TRO-001DR8\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003ePaludibacterium yongneupense\u003c/em\u003e 5YN8-15\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003ePseudogulbenkiania subflava\u003c/em\u003e DSM 22618\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eGulbenkiania mobilis\u003c/em\u003e E4FC31\u003csup\u003eT\u003c/sup\u003e, were cultured on R2A agar at 25\u0026deg;C for 2 days for comparison. \u003cem\u003eCraterilacusibacter sinensis\u003c/em\u003e B2N2-7\u003csup\u003eT\u003c/sup\u003e was cultivated in PYG medium. Enzymatic and physiological activities were determined using a set of enzymatic tests API ZYM and 20NE (BioM\u0026eacute;rieux), and acids production was tested using API 50CH (BioM\u0026eacute;rieux) strips. Carbon sources utilization was measured using the GEN III MicroPlate system (Biolog) following the manufacturer\u0026rsquo;s protocol.\u003c/p\u003e\n\u003ch3\u003eChemotaxonomic Analyses\u003c/h3\u003e\n\u003cp\u003eStrain TC5R-5\u003csup\u003eT\u003c/sup\u003e was cultured in R2A broth at 25\u0026deg;C with 180 rpm for 48h and harvested during the late exponential phase. Cells were collected for further analysis of fatty acids, respiratory quinones, and polar lipids. Fatty acids were extracted following the standard protocol by Miller and Kuykendall and analysed using the Sherlock Microbial Identification System (MIDI) [Miller, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e1982\u003c/span\u003e; Sasser, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e1990\u003c/span\u003e]. The collected cells were freeze-dried for chemotaxonomic analysis. Polar lipids were extracted as per the protocol outlined by Komagata and Suzuki (\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e1987\u003c/span\u003e) [Komagata and Suzuki, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e1987\u003c/span\u003e] and separated using two-dimensional silica gel thin layer chromatography. The first direction involved a mixture of chloroform: methanol:water (65:25:4, v/v/v), while the second direction used a mixture of chloroform : acetic acid : methanol : water (80:15:12:4, v/v/v/v). Finally, molybdatophosphoric acid, molybdenum blue, ninhydrin and anisaldehyde reagent were utilized to detect total polar lipid, phospholipids, aminolipids, and glycolipids [Minnikin et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e1993\u003c/span\u003e]. The specific components of respiratory quinones were extracted, separated, purified and analysed by HPLC following the previously described approach [ Collins, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e1985\u003c/span\u003e; Wu et al., \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e1989\u003c/span\u003e].\u003c/p\u003e"},{"header":"Results and Discussion","content":"\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003ePhylogenetic and phylogenomic analysis\u003c/h2\u003e\u003cp\u003eBased on 16S rRNA gene sequence comparisons, strain TC5R-5\u003csup\u003eT\u003c/sup\u003e exhibited the highest similarity (94.3%) to \u003cem\u003eChromobacterium alkanivorans\u003c/em\u003e IITR-71\u003csup\u003eT\u003c/sup\u003e [Bajaj et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2016\u003c/span\u003e]. Phylogenetic analysis using the maximum-likelihood method with the GTR\u0026thinsp;+\u0026thinsp;G\u0026thinsp;+\u0026thinsp;I nucleotide substitution model placed strain TC5R-5\u003csup\u003eT\u003c/sup\u003e within a clade containing the genera \u003cem\u003eChromobacterium\u003c/em\u003e, \u003cem\u003eAquitalea\u003c/em\u003e, \u003cem\u003eGulbenkiania\u003c/em\u003e, and \u003cem\u003ePseudogulbenkiania\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Consistent with this, phylogenetic trees reconstructed using neighbor-joining and maximum-parsimony methods also supported the close affiliation of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e with members of the family \u003cem\u003eChromobacteriaceae\u003c/em\u003e (order \u003cem\u003eNeisseriales\u003c/em\u003e). The topological structures of the neighbor-joining and maximum-parsimony trees were congruent with that of the maximum-likelihood tree (see Supplementary Material Figure \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eTo further clarify the phylogenetic position of strain TC5R-5T, phylogenomic trees were reconstructed using TYGS, IQ-TREE, and UBGC. The maximum-likelihood tree generated with IQ-TREE v2.1.2 under the MFP model and 1000 ultrafast bootstrap replicates, with selected species from \u003cem\u003eNeisseriales\u003c/em\u003e and \u003cem\u003eBetaproteobacteria\u003c/em\u003e as outgroups, placed strain TC5R-5\u003csup\u003eT\u003c/sup\u003e within a cluster comprising \u003cem\u003eChromobacterium\u003c/em\u003e species. This branch was neighbored by the genera \u003cem\u003eAquitalea\u003c/em\u003e, \u003cem\u003eGulbenkiania\u003c/em\u003e, \u003cem\u003ePaludibacterium\u003c/em\u003e, and \u003cem\u003ePseudogulbenkiania\u003c/em\u003e, all belonging to the family \u003cem\u003eChromobacteriaceae\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ea). A similar phylogenetic placement was supported by the bacterial core gene-based tree (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eb), in which strain TC5R-5\u003csup\u003eT\u003c/sup\u003e reliably grouped with members of the \u003cem\u003eChromobacteriaceae\u003c/em\u003e, including species of \u003cem\u003ePaludibacterium\u003c/em\u003e, \u003cem\u003eAquitalea\u003c/em\u003e, \u003cem\u003eChromobacterium\u003c/em\u003e, \u003cem\u003eVogesella\u003c/em\u003e, and \u003cem\u003eGulbenkiania\u003c/em\u003e, among others. These results suggest that strain TC5R-5\u003csup\u003eT\u003c/sup\u003e may represent a novel species within this family. In contrast, the TYGS-based phylogenomic tree revealed a divergent topology (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ec). Here, strain TC5R-5\u003csup\u003eT\u003c/sup\u003e clustered with \u0026lsquo;\u003cem\u003eHalomonas humidisoli\u003c/em\u003e\u0026rsquo;, \u003cem\u003eVreelandella glaciei\u003c/em\u003e, and \u003cem\u003eAndreprevotia sp\u003c/em\u003e. However, based on physiological and ecological characteristics, the strain shows no clear affiliation with \u003cem\u003eHalomonas\u003c/em\u003e species. Notably, it shares certain similarities with \u003cem\u003eAndreprevotia chitinilytica\u003c/em\u003e JS11-7\u003csup\u003eT\u003c/sup\u003e, a member of the family \u003cem\u003eChitinibacteraceae\u003c/em\u003e.\u003c/p\u003e\u003cp\u003eThe ANI and dDDH values between strain TC5R-5\u003csup\u003eT\u003c/sup\u003e and its closest relative, \u003cem\u003eChromobacterium alkanivorans\u003c/em\u003e IITR-71\u003csup\u003eT\u003c/sup\u003e, were 73.5% and 13.6%, respectively\u0026mdash;both well below the established species demarcation thresholds of 95\u0026ndash;96% for ANI and 70% for dDDH. Additionally, the AAI value between the two strains was 52.8%, which fell beneath the proposed genus boundary range of 60\u0026ndash;80% [Luo et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2014\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eTo further evaluate its taxonomic position, ANI values were compared between strain TC5R-5\u003csup\u003eT\u003c/sup\u003e and several reference strains: the phylogenetically closest relatives within the \u003cem\u003eChromobacteriaceae\u003c/em\u003e (based on 16S rRNA and IQ-TREE/UBGC phylogenomic trees), \u003cem\u003eChromobacterium alkanivorans\u003c/em\u003e IITR-71\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eAquitalea palustris\u003c/em\u003e CCM7557\u003csup\u003eT\u003c/sup\u003e, as well as the top hits from the TYGS-based tree, including \u003cem\u003eAndreprevotia chitinilytica\u003c/em\u003e DSM 18519\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eHalomonas humidisoli\u003c/em\u003e WN018\u003csup\u003eT\u003c/sup\u003e, and \u003cem\u003eVreelandella glaciei\u003c/em\u003e DD39\u003csup\u003eT\u003c/sup\u003e (Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). The ANI values between strain TC5R-5\u003csup\u003eT\u003c/sup\u003e and the \u003cem\u003eChromobacteriaceae\u003c/em\u003e members \u003cem\u003eC. alkanivorans\u003c/em\u003e IITR-71\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eA. palustris\u003c/em\u003e CCM7557\u003csup\u003eT\u003c/sup\u003e (73.48%, 72.49%) were notably higher than those with \u003cem\u003eA. chitinilytica\u003c/em\u003e DSM 18519\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eH. humidisoli\u003c/em\u003e WN018\u003csup\u003eT\u003c/sup\u003e, and \u003cem\u003eV. glaciei\u003c/em\u003e DD39\u003csup\u003eT\u003c/sup\u003e (68.49%, 64.26%, 64.73%). These comparisons were also supported by substantially greater genome coverage (26.78% and 25.03% vs. 9.92%, 5.01%, and 4.91%, respectively). Together, these results corroborate that strain TC5R-5\u003csup\u003eT\u003c/sup\u003e is phylogenetically more closely affiliated with the \u003cem\u003eChromobacteriaceae\u003c/em\u003e.\u003c/p\u003e\u003cp\u003eIn summary, integrated phylogenetic analyses based on both 16S rRNA gene and whole-genome sequences support the classification of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e as a novel species of a new genus within the order \u003cem\u003eNeisseriales\u003c/em\u003e. Although some discrepancies were observed among phylogenomic trees constructed using different methods, current evidence indicates that its placement within the family \u003cem\u003eChromobacteriaceae\u003c/em\u003e is the most appropriate and rational provisional assignment.\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eGenome Features and Biosynthetic Gene Cluster\u003c/h3\u003e\n\u003cp\u003eThe complete genome of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e comprises 3.44 Mb with a total of 3408 genes, including 3274 protein-coding sequences (CDS). The genomic DNA G\u0026thinsp;+\u0026thinsp;C content, derived from sequencing data, was determined to be 51.4%. A comparative summary of genomic features between strain TC5R-5\u003csup\u003eT\u003c/sup\u003e and its phylogenetic relatives is provided in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. Annotation via RAST revealed that 29% of the chromosomal genes could be assigned to functional categories, distributed across 291 subsystems within 27 broader functional groups. The most abundant subsystems were \u0026lsquo;Amino Acids and Derivatives\u0026rsquo;, \u0026lsquo;Protein Metabolism\u0026rsquo;, and \u0026lsquo;Cofactors, Vitamins, Prosthetic Groups, Pigments\u0026rsquo; (Figure S2). AntiSMASH analysis identified a biosynthetic gene cluster (BGC) located on scaffold 7, spanning 24,974 bp, which showed high similarity to the rhizomide BGC from \u003cem\u003eParaburkholderia rhizoxinica\u003c/em\u003e HKI 454 (NC_014718.1; Figure S3). According to MIBiG annotation, this cluster is predicted to encode a nonribosomal peptide synthetase (NRPS) consisting of seven modules and containing a starter condensation domain, suggesting the potential production of a lipopeptide-class compound.\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\u003eComparison of genome features between strain TC5R-5\u003csup\u003eT\u003c/sup\u003e and the type species of genus in the family Strains: 1. TC5R-5\u003csup\u003eT\u003c/sup\u003e (JACCFC000000000); 2. \u003cem\u003eChromobacterium alkanivorans\u003c/em\u003e IITR-71\u003csup\u003eT\u003c/sup\u003e (JAFHKL000000000); 3. \u003cem\u003eChromobacterium violaceum\u003c/em\u003e ATCC 12472\u003csup\u003eT\u003c/sup\u003e (AE016825); 4. \u003cem\u003eAquitalea magnusonii\u003c/em\u003e DSM 25134\u003csup\u003eT\u003c/sup\u003e (QJKC00000000); 5. \u003cem\u003ePaludibacterium yongneupense\u003c/em\u003e 5YN8-15\u003csup\u003eT\u003c/sup\u003e (AUGZ00000000); 6. \u003cem\u003ePseudogulbenkiania subflava\u003c/em\u003e DSM 22618\u003csup\u003eT\u003c/sup\u003e (FXAG00000000); 7. \u003cem\u003eGulbenkiania mobilis\u003c/em\u003e E4FC31\u003csup\u003eT\u003c/sup\u003e (SMDA00000000); 8. \u003cem\u003eCraterilacuibacter sinensis\u003c/em\u003e B2N2-7\u003csup\u003eT\u003c/sup\u003e (WSSB00000000); 9. \u003cem\u003eCrenobacter luteus\u003c/em\u003e YIM 78141\u003csup\u003eT\u003c/sup\u003e (SLXG00000000); 10. \u003cem\u003eVogesella indigofera\u003c/em\u003e DSM 3303\u003csup\u003eT\u003c/sup\u003e (RBID00000000).\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"11\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003e6\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003e8\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c10\"\u003e\u003cp\u003e9\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c11\"\u003e\u003cp\u003e10\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\u003eGenome size (Mb)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e4.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4.4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e4.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e4.4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e3.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e2.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e3.6\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eGC content (%)\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e51.4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e64.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e64.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e60.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e62.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e63.4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e63.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e59.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e68.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e64.5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eNumber of genes\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3408\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4839\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e4529\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4198\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e4125\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e4064\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2805\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e3153\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e2845\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e3454\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eNumber of CDS\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3274\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4733\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e4407\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4099\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e4010\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e3979\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2717\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e3041\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e2760\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e3333\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\n\u003ch3\u003ePhenotypic and Physiological Characteristics\u003c/h3\u003e\n\u003cp\u003eColonies of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e were white and non-transparent. The cells were rod-shaped, measuring approximately 0.8\u0026ndash;0.9 \u0026micro;m in width and 3.0\u0026ndash;4.0 \u0026micro;m in length, and exhibited a single polar flagellum (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Cells were Gram-stain-negative, catalase- and oxidase-positive, but did not display DNase activity. Growth occurred only under aerobic conditions. The strain tolerated NaCl concentrations ranging from 0 to 0.5% (w/v), and was capable of growth at temperatures between 15\u0026ndash;30\u0026deg;C and pH 6.0\u0026ndash;8.0, with optimal growth observed at 20\u0026ndash;30\u0026deg;C and pH 6.0\u0026ndash;7.0.\u003c/p\u003e\u003cp\u003eSubstrate degradation assays were performed on R2A agar supplemented with various compounds (5.0% casein, 1.0% aesculin, 1.0% Tween 20, 1.0% Tween 80, 1.0% starch; w/v). Strain TC5R-5\u003csup\u003eT\u003c/sup\u003e demonstrated limited carbon source utilization and acid production capabilities. It also tested negative for numerous enzymatic activities and biochemical reactions when compared to type strains of related genera within the family \u003cem\u003eChromobacteriaceae\u003c/em\u003e. A summary of its morphological and physiological characteristics is provided in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\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\u003eDifferential characteristics between strain TC5R-5\u003csup\u003eT\u003c/sup\u003e and most related phylogenetic neighbors Strains: 1. TC5R-5\u003csup\u003eT\u003c/sup\u003e; 2. \u003cem\u003eChromobacterium alkanivorans\u003c/em\u003e IITR-71\u003csup\u003eT\u003c/sup\u003e; 3. \u003cem\u003eChromobacterium violaceum\u003c/em\u003e ATCC 12472\u003csup\u003eT\u003c/sup\u003e; 4. \u003cem\u003eAquitalea magnusonii\u003c/em\u003e TRO-001DR8\u003csup\u003eT\u003c/sup\u003e; 5. \u003cem\u003ePaludibacterium yongneupense\u003c/em\u003e 5YN8-15\u003csup\u003eT\u003c/sup\u003e; 6. \u003cem\u003ePseudogulbenkiania subflava\u003c/em\u003e DSM 22618\u003csup\u003eT\u003c/sup\u003e; 7. \u003cem\u003eGulbenkiania mobilis\u003c/em\u003e E4FC31\u003csup\u003eT\u003c/sup\u003e; 8. \u003cem\u003eCraterilacusibacter sinensis\u003c/em\u003e B2N2-7\u003csup\u003eT\u003c/sup\u003e; 9. \u003cem\u003eCrenobacter luteus\u003c/em\u003e YIM 78141\u003csup\u003eT\u003c/sup\u003e; 10. \u003cem\u003eVogesella indigofera\u003c/em\u003e DSM 3303\u003csup\u003eT\u003c/sup\u003e. +, Positive; -, negative; w, weakly positive. All of the data are acquired from this experiment except where indicated otherwise. \u003csup\u003ea,b,c,d,e,f\u003c/sup\u003e Data from corresponding literatures [Liu et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Dong et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Subhash et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Sedl\u0026aacute;ček et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Sheu et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Vaz-Moreira et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2007\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"11\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003e6\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003e8\u003csup\u003ea,\u003c/sup\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c10\"\u003e\u003cp\u003e9\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c11\"\u003e\u003cp\u003e10\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eReactions (API 20E\u0026amp;20NE)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eFermentation of glucose\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\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\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eArginine dihydrolase\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\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\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHydrolysis of gelatin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\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\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eEnzyme activity (API ZYM )\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\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eα\u003c/em\u003e-Chymotrypsin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\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\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eN-Acetyl-\u003cem\u003eβ\u003c/em\u003e- glucosaminidase\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\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\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003ew\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eUtilization of substrates (Biolog GENIII)\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\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eD-Trehalose\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ew\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ew\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eInosine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\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\u003ew\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGlycerol\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\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\u003ew\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGelatin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\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\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eL-Arginine\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ew\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003ew\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003ew\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePectin\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ew\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\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003ew\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eMucic acid\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ew\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\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003ew\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eα \u0026ndash;Keto-glutaric acid\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003ew\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003ew\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003ew\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eFormic acid\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e+\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\u003ew\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003ew\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003ew\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eAcids production (API 50CH)\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\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eD-Glucose\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e+\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\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003ew\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003ew\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eD- Mannose\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\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\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTrehalose\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\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\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003eRespiratory quinone\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eQ-8, Q-7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eQ-8, Q-7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eQ-8\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eQ-8\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eQ-8\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eQ-8, Q-9\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003eQ-8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003eQ-8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cb\u003ePolar Lipid\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\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePE\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e+\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\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e+\u003csup\u003e[e]\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e+\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e+\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\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDPG\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e+\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\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e+\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAPL\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAL\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePL\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1\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\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eL\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\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\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAGP\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\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\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eND\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003eChemotaxonomic Characteristics\u003c/h2\u003e\u003cp\u003eThe cellular fatty acid profile of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e was presented in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e. The predominant component was summed feature 3 (C\u003csub\u003e16:1\u003c/sub\u003e \u003cem\u003eω\u003c/em\u003e7\u003cem\u003ec\u003c/em\u003e/C\u003csub\u003e16:1\u003c/sub\u003e \u003cem\u003eω\u003c/em\u003e6\u003cem\u003ec\u003c/em\u003e; 51.6%), which was notably higher than those observed in related reference strains. In contrast, the content of summed feature 8 (C\u003csub\u003e18:1\u003c/sub\u003e \u003cem\u003eω\u003c/em\u003e7c/C\u003csub\u003e18:1\u003c/sub\u003e \u003cem\u003eω\u003c/em\u003e6c) in strain TC5R-5\u003csup\u003eT\u003c/sup\u003e was less than half of that detected in \u003cem\u003eChromobacterium alkanivorans\u003c/em\u003e IITR-71\u003csup\u003eT\u003c/sup\u003e. Overall, the principal fatty acids in strain TC5R-5\u003csup\u003eT\u003c/sup\u003e and all type strains of the family \u003cem\u003eChromobacteriaceae\u003c/em\u003e were summed were summed feature 3 (C\u003csub\u003e16:1\u003c/sub\u003e \u003cem\u003eω\u003c/em\u003e7\u003cem\u003ec\u003c/em\u003e/C\u003csub\u003e16:1\u003c/sub\u003e \u003cem\u003eω\u003c/em\u003e6\u003cem\u003ec\u003c/em\u003e) and C\u003csub\u003e16:0\u003c/sub\u003e. The major polar lipids of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e were phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), one unidentified aminophospholipid (APL1), and one unidentified phospholipid (PL1) (Figure S4). For comparative classification, the polar lipid profile of \u003cem\u003eChromobacterium alkanivorans\u003c/em\u003e IITR-71\u003csup\u003eT\u003c/sup\u003e was also examined. The results indicated the presence of four aminolipids and one aminophospholipid in \u003cem\u003eChromobacterium alkanivorans\u003c/em\u003e IITR-71\u003csup\u003eT\u003c/sup\u003e, which were absent in TC5R-5\u003csup\u003eT\u003c/sup\u003e. Moreover, the comparison between strain TC5R-5\u003csup\u003eT\u003c/sup\u003e and the type species of related genera in the family \u003cem\u003eChromobacteriaceae\u003c/em\u003e were also conducted. As shown in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, polar lipids of most of type strains, except \u003cem\u003eCrenobacter luteus\u003c/em\u003e YIM 78141\u003csup\u003eT\u003c/sup\u003e, consisted of PE, PG and DPG. The predominant respiratory quinone of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e was ubiquinone 8 (Q-8), being in line with the quinone profile of members of the family \u003cem\u003eChromobacteriaceae\u003c/em\u003e.\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\u003eWhole-cell fatty acid profiles of the novel isolates and the type strains of related species of genera 1.strain TC5R-5\u003csup\u003eT\u003c/sup\u003e; 2. \u003cem\u003eChromobacterium alkanivorans\u003c/em\u003e IITR-71\u003csup\u003eT\u003c/sup\u003e; 3. \u003cem\u003eChromobacterium violaceum\u003c/em\u003e ATCC 12472\u003csup\u003eT\u003c/sup\u003e; 4. \u003cem\u003eAquitalea magnusonii\u003c/em\u003e TRO-001DR8\u003csup\u003eT\u003c/sup\u003e; 5. \u003cem\u003ePaludibacterium yongneupense\u003c/em\u003e 5YN8-15\u003csup\u003eT\u003c/sup\u003e; 6. \u003cem\u003ePseudogulbenkiania subflava\u003c/em\u003e DSM 22618\u003csup\u003eT\u003c/sup\u003e; 7. \u003cem\u003eGulbenkiania mobilis\u003c/em\u003e E4FC31\u003csup\u003eT\u003c/sup\u003e; 8. \u003cem\u003eCraterilacusibacter sinensis\u003c/em\u003e B2N2-7\u003csup\u003eT\u003c/sup\u003e; 9. \u003cem\u003eCrenobacter luteus\u003c/em\u003e YIM 78141\u003csup\u003eT\u003c/sup\u003e; 10. \u003cem\u003eVogesella indigofera\u003c/em\u003e DSM 3303\u003csup\u003eT\u003c/sup\u003e. All data were from this study unless otherwise indicated. *Summed features are groups of two acids that cannot be separated by GLC using the MIDI system. \u003csup\u003ea\u003c/sup\u003e Data from Lau et al., \u003csup\u003eb\u003c/sup\u003e Data from Liu et al., \u003csup\u003ec\u003c/sup\u003e Data from Dong et al.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"11\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003e6\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003e8\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c10\"\u003e\u003cp\u003e9\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c11\"\u003e\u003cp\u003e10\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eC\u003csub\u003e12:0\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e7.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e10.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e5.6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e7.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e2.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e6.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e3.6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e2.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e5.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e3.8\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eC\u003csub\u003e14:0\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e2.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e3.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e1.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e1.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e3.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e0.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e3.6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e0.5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eC\u003csub\u003e16:0\u003c/sub\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e26.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e20.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e23.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e30.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e29.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e28.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e24.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e33.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e27.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e24.9\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eC\u003csub\u003e10:0\u003c/sub\u003e 3-OH\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e3.6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e4.6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e5.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e3.6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e3.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e3.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e2.9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e2.6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e7.2\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eC\u003csub\u003e12:0\u003c/sub\u003e 3-OH\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e2.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e5.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e2.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e3.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e2.6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e3.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e3.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e2.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSummed feature 3*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e51.6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e44.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e35.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e40.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e34.4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e38.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e43.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e31.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e44.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e45.6\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSummed feature 8*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e\u003cp\u003e5.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e12.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e15.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e10.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e13.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e11.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e9.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e1.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c11\"\u003e\u003cp\u003e7.9\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"11\"\u003e* Summed Features are fatty acids that cannot be resolved reliably from another fatty acid using the chromatographic conditions chosen. The MIDI system groups these fatty acids together as one feature with a single percentage of the total. Summed feature 3, C\u003csub\u003e16:1\u003c/sub\u003e \u003cem\u003eω\u003c/em\u003e7\u003cem\u003ec\u003c/em\u003e/C\u003csub\u003e16:1\u003c/sub\u003e \u003cem\u003eω\u003c/em\u003e6\u003cem\u003ec\u003c/em\u003e; Summed feature 8, C\u003csub\u003e18:1\u003c/sub\u003e \u003cem\u003eω\u003c/em\u003e7\u003cem\u003ec\u003c/em\u003e/C\u003csub\u003e18:1\u003c/sub\u003e \u003cem\u003eω\u003c/em\u003e6\u003cem\u003ec\u003c/em\u003e. Data from corresponding literatures.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003eTaxonomic Conclusion\u003c/h2\u003e\u003cp\u003eWe conducted comprehensive phylogenetic analysis based on the 16S rRNA gene and whole genome alignments to determine the phylogenetic position of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003ePhylogenetic trees reconstructed from the 16S rRNA gene consistently placed the strain within the family \u003cem\u003eChromobacteriaceae\u003c/em\u003e. This assignment was further supported by phylogenomic analyses performed with IQ-TREE using single-copy genes and with UBGC based on core genes. In contrast, the phylogenomic tree generated with the TYGS online tool showed that strain TC5R-5ᵀ formed a cluster with \u003cem\u003eHalomonas humidisoli\u003c/em\u003e WN018ᵀ, \u003cem\u003eVreelandella glaciei\u003c/em\u003e DD39ᵀ, and \u003cem\u003eAndreprevotia chitinilytica\u003c/em\u003e JS11-7ᵀ.\u003c/p\u003e\u003cp\u003eSubsequently, we calculated the Average Nucleotide Identity (ANI) and genome coverage between strain TC5R-5\u003csup\u003eT\u003c/sup\u003e with its phylogenetic neighbors identified in the phylogenomic trees (Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). The results showed that strain TC5R-5\u003csup\u003eT\u003c/sup\u003e shared higher ANI values with \u003cem\u003eChromobacterium alkanivorans\u003c/em\u003e IITR-71\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eAquitalea palustris\u003c/em\u003e CCM7557\u003csup\u003eT\u003c/sup\u003e (73.48%, 72.49%) than with \u003cem\u003eAndreprevotia chitinilytica\u003c/em\u003e DSM 18519\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eHalomonas humidisoli\u003c/em\u003e WN018\u003csup\u003eT\u003c/sup\u003e, and \u003cem\u003eVreelandella glaciei\u003c/em\u003e DD39\u003csup\u003eT\u003c/sup\u003e (68.49%, 64.26%, 64.73%). Genome coverage was also notably greater with the former group (26.78% and 25.03%, respectively) compared to the latter (9.92%, 5.01%, and 4.91%, respectively). These findings further corrobrorated that strain TC5R-5ᵀ is phylogenetically affiliated with the family \u003cem\u003eChromobacteriaceae\u003c/em\u003e.\u003c/p\u003e\u003cp\u003eIn summary, comprehensive phylogenetic analyses based on the 16S rRNA gene and whole-genome sequences consistently support the classification of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e as representing a novel genus and species within the order \u003cem\u003eNeisseriales\u003c/em\u003e. Although minor topological variations were observed among phylogenomic trees constructed using different analytical methods, the overall evidence strongly supported its provisional classification within the family \u003cem\u003eChromobacteriaceae\u003c/em\u003e as the most coherent and biologically plausible placement.\u003c/p\u003e\u003cp\u003e\u003cb\u003eDescription of\u003c/b\u003e \u003cb\u003eCraterilacuimicrobium\u003c/b\u003e \u003cb\u003egen. nov.\u003c/b\u003e\u003c/p\u003e\u003cp\u003e\u003cem\u003eCraterilacuimicrobium\u003c/em\u003e gen. nov. (Cra.te.ri.la.cu.i.mi.cro'bium. L. masc. n. \u003cem\u003ecrater\u003c/em\u003e, a crater; L. masc. n. \u003cem\u003elacus\u003c/em\u003e, a lake; N.L. neut. n. \u003cem\u003emicrobium\u003c/em\u003e, a microbe; N.L. neut. n. \u003cem\u003eCraterilacuimicrobium\u003c/em\u003e, a microbe from a crater lake)\u003c/p\u003e\u003cp\u003eCells rod-shaped, motile, Gram-stain-negative and strictly aerobic. Catalase- and oxidase-positive. The major fatty acids consist of summed feature 3 (C\u003csub\u003e16:1\u003c/sub\u003e \u003cem\u003eω\u003c/em\u003e7\u003cem\u003ec\u003c/em\u003e/C\u003csub\u003e16:1\u003c/sub\u003e \u003cem\u003eω\u003c/em\u003e6\u003cem\u003ec\u003c/em\u003e). The predominant ubiquinone is Q-8, with a small amount of Q-7. The main polar lipids include phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), one unidentified aminophospholipid (APL), and one unidentified phospholipid (PL). The type species is \u003cem\u003eCraterilacuimicrobium aquaticum\u003c/em\u003e.\u003c/p\u003e\u003cp\u003e\u003cb\u003eDescription of\u003c/b\u003e \u003cb\u003eCraterilacuimicrobium aquaticum\u003c/b\u003e \u003cb\u003esp. nov.\u003c/b\u003e\u003c/p\u003e\u003cp\u003e\u003cem\u003eCraterilacuimicrobium aquaticum\u003c/em\u003e sp. nov. (a.qua'ti.cum. L. neut. adj. \u003cem\u003eaquaticum\u003c/em\u003e, living, growing or found in or by water, aquatic)\u003c/p\u003e\u003cp\u003eCells are Gram-stain-negative, strictly aerobic, motile, and short rod-shaped. Colonies are circular, smooth, white-colored after incubating for 48 h at 25\u0026deg;C on R2A agar. Growth occurs at 15\u0026ndash;30\u0026deg;C (optimum, 20\u0026ndash;30\u0026deg;C), at pH 6.0\u0026ndash;8.0 (optimum, 6.0\u0026ndash;7.0) and with 0-0.5% NaCl (optimum, 0-0.5%, w/v). Catalase- and oxidase-activity present. DNase activity is not present. Positive for hydrolyzation of Tween 20, but negative for degradation of casein, cellulose, starch, aesculin and Tween 80. In the API 20 NE and 20E tests, strain TC5R-5\u003csup\u003eT\u003c/sup\u003e could reduce nitrate to nitrite. Voges-Proskauer-test and fermentation of glucose are positive. Citrate not utilized, indole not produced, no arginine dihydrolase activity, no hydrolysis of gelatin and esculin. Does not produce H\u003csub\u003e2\u003c/sub\u003eS. The activities of ornithine decarboxylase, lysine decarboxylase, tryptophane deaminase, urease, ortho-nitrophenyl-\u003cem\u003eβ\u003c/em\u003e-D-galactopyranosidase and para-nitrophenyl-\u003cem\u003eβ\u003c/em\u003e-D-galactopyranosidase are negative. In the API ZYM test, positive for alkaline phosphatase, leucine arylamidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase, but negative for esterase (C4), esterase lipase (C8), lipase (C14), valine arylamidase, cystine arylamidase, trypsin, \u003cem\u003eα\u003c/em\u003e-chymotrypsin, \u003cem\u003eα\u003c/em\u003e-galactosidase, \u003cem\u003eβ\u003c/em\u003e-galactosidase, \u003cem\u003eβ\u003c/em\u003e-glucuronidase, \u003cem\u003eα\u003c/em\u003e-glucosidase, \u003cem\u003eβ\u003c/em\u003e-glucosidase, \u003cem\u003eN\u003c/em\u003e-acetyl-\u003cem\u003eβ\u003c/em\u003e-glucosaminidase, \u003cem\u003eα\u003c/em\u003e-mannosidase and \u003cem\u003eα\u003c/em\u003e-fucosidase. Acids could be produced from glucose in API 50CH, but not other carbon sources. In the BIOLOG GEN III MicroPlate system, \u003cem\u003eα\u003c/em\u003e-D-glucose, D-glucose-6-PO\u003csub\u003e4\u003c/sub\u003e, L-alanin, L-glutamic acid, L-histidine, L-serine, methyl pyruvate, L-lactic acid, \u003cem\u003eα\u003c/em\u003e-keto-glutaric acid, \u003cem\u003eγ\u003c/em\u003e-amino-butrytic acid, \u003cem\u003eβ\u003c/em\u003e\u0026ndash;hydroxyl-D, L-butyric acid, acetoacetic acid, formic acid are utilized as sole carbon resource, and dextrin, D-frucose, D-fucose, L-fucose, D-fructose-6-PO\u003csub\u003e4\u003c/sub\u003e, D-serine, glycyl-L-proline, L-aspartic acid, L-pyroglutamic acid, pectin, D-galacturonic acid, D-glucuronic acid, glucuronamide, mucic acid, D-lactic acid metyl ester, citric acid, D-malic acid, L-malic acid, Tween 40, \u003cem\u003eα\u003c/em\u003e-hydroxy-butyric acid, propionic acid, acetic acid are weakly utilized. The genomic DNA G\u0026thinsp;+\u0026thinsp;C content is 51.4% according to the genomic sequencing data. The major fatty acids (\u0026gt;\u0026thinsp;10%) are summed feature 3 (C\u003csub\u003e16:1\u003c/sub\u003e \u003cem\u003eω\u003c/em\u003e7\u003cem\u003ec\u003c/em\u003e/C\u003csub\u003e16:1\u003c/sub\u003e \u003cem\u003eω\u003c/em\u003e6\u003cem\u003ec\u003c/em\u003e) and C\u003csub\u003e16:0\u003c/sub\u003e. The predominant respiratory quinone is Q-8 with a trace amount of Q-7. The main polar lipids are phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), one unidentified aminophospholipid (APL) and one unidentified phospholipid (PL).\u003c/p\u003e\u003cp\u003eThe type strain TC5R-5\u003csup\u003eT\u003c/sup\u003e (=\u0026thinsp;CGMCC 1.17058\u003csup\u003eT\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;KCTC 72736\u003csup\u003eT\u003c/sup\u003e ) was isolated from water of Aershan Tianchi crater lake in the Inner Mongolia Autonomous Region of China. The GenBank/EMBL/DDBJ accession number for 16S rRNA gene of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e is OK487579 and the DDBJ/ENA/GenBank accession number of the Whole Genome Shotgun project of TC5R-5\u003csup\u003eT\u003c/sup\u003e is JACCFC000000000.\u003c/p\u003e\u003c/div\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eANI, average nucleotide identity; APL, unidentified aminophospholipid; CDS, coding sequences; DPG, diphosphatidylglycerol; PE, phosphatidylethanolamine; PG, phosphatidylglycerol; PL, unidentified phospholipid; AGP, aminoglycophospholipid; L, unidentified polar lipid; NRPS, nonribosomal peptide synthetase.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eConflicts of interest:\u003c/strong\u003e\u003cp\u003eThe authors declare that there are no conflicts of interest.\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eEthical approval:\u003c/strong\u003e\u003cp\u003eThe data reported in this article is not derived from any studies or tests on human volunteers or animals.\u003c/p\u003e\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eInformed consent:\u003c/strong\u003e\u003cp\u003eAll the authors have read and approved the final manuscript for submission.\u003c/p\u003e\u003c/p\u003e\u003ch2\u003eFunding information\u003c/h2\u003e\u003cp\u003eThis work was financially supported by the National Natural Science Foundation of China (31970004), the Strategic Biological Resources Capacity Building Project of Chinese Academy of Sciences, China (KFJ-BRP-017-73) and the Key Research and Development Project of Xizang Autonomous Region, China (XZ202401ZY0061).\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eYou-Jun Liao: Conceptualization, Resources, Data curation, Methodology; Xuan Zhang : Investigation, Formal analysis; Zi-Xuan Liu: Visualization, Writing\u0026mdash;Original Draft and Editing; Ai-Hua Li: Funding acquisition, Project administration, Writing\u0026mdash;Reviewing.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eThe authors gratefully acknowledge the help from Prof. Teun Boekhout (College of Sciences, King Saud University, Saudi Arabia) to improve the English text, the help from Dr. Jing-Nan Liang for the TEM observation, and the assistance from Prof. Stefano Ventura (Research institute on Terrestrial Ecosystems, Firenze Unit, Italy) for his advice on bacterial nomenculature.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eThe GenBank/EMBL/DDBJ accession number for 16S rRNA gene of strain TC5R-5T is OK487579. The Whole Genome Shotgun project of strain TC5R-5T has been deposited at DDBJ/ENA/GenBank under the accession number JACCFC000000000. This strain is available at China General Microbiology Culture Collection Center (CGMCC) under the accession no. CGMCC 1.17058T.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAdeolu M, Gupta RS (2013) Phylogenomics and molecular signatures for the order \u003cem\u003eNeisseriales\u003c/em\u003e: proposal for division of the order \u003cem\u003eNeisseriales\u003c/em\u003e into the emended family \u003cem\u003eNeisseriaceae\u003c/em\u003e and \u003cem\u003eChromobacteriaceae\u003c/em\u003e fam. nov. \u003cem\u003eAntonie Van Leeuwenhoek\u003c/em\u003e 104:1\u0026ndash;24. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s10482-013-9920-6\u003c/span\u003e\u003cspan address=\"10.1007/s10482-013-9920-6\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAziz RK, Bartels D, Best AA \u003cem\u003eet al\u003c/em\u003e (2008) The rast server: rapid annotations using subsystems technology. \u003cem\u003eBMC Genomics\u003c/em\u003e 9:75\u0026ndash;75. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/1471-2164-9-75\u003c/span\u003e\u003cspan address=\"10.1186/1471-2164-9-75\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBajaj A, Kumar A, Yadav S, Kaur G, Bala M \u003cem\u003eet al.\u003c/em\u003e (2016) Isolation and characterization of a novel Gram-negative bacterium \u003cem\u003eChromobacterium alkanivorans\u003c/em\u003e sp. nov., strain IITR-71\u003csup\u003eT\u003c/sup\u003e degrading halogenated alkanes. \u003cem\u003eInt J Syst Evol Microbiol\u003c/em\u003e 66:5228\u0026ndash;5235. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijsem.0.001500\u003c/span\u003e\u003cspan address=\"10.1099/ijsem.0.001500\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChen S, Rudra B, Gupta RS (2021) Phylogenomics and molecular signatures support division of the order \u003cem\u003eNeisseriales\u003c/em\u003e into emended families \u003cem\u003eNeisseriaceae\u003c/em\u003e and \u003cem\u003eChromobacteriaceae\u003c/em\u003e and three new families \u003cem\u003eAquaspirillaceae\u003c/em\u003e fam. nov., \u003cem\u003eChitinibacteraceae\u003c/em\u003e fam. nov., and \u003cem\u003eLeeiaceae\u003c/em\u003e fam. nov. \u003cem\u003eSyst Appl Microbiol\u003c/em\u003e 44:126251. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.syapm.2021.126251\u003c/span\u003e\u003cspan address=\"10.1016/j.syapm.2021.126251\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChun J, Oren A, Ventosa A \u003cem\u003eet al\u003c/em\u003e (2018) Proposed minimal standards for the use of genome data for the taxonomy of prokaryotes. \u003cem\u003eInt J Syst Evol Microbiol\u003c/em\u003e 68:461\u0026ndash;466. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijsem.0.002516\u003c/span\u003e\u003cspan address=\"10.1099/ijsem.0.002516\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChou JH, Chou YJ, Arun AB, et al (2009) \u003cem\u003eVogesella lacus\u003c/em\u003e sp. nov., isolated from a soft-shell turtle culture pond. \u003cem\u003eInt J Syst Evol Microbiol\u003c/em\u003e 59:2629\u0026ndash;2632. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijs.0.009266-0\u003c/span\u003e\u003cspan address=\"10.1099/ijs.0.009266-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCollins MD (1985) Isoprenoid quinone analysis in classification and identification. In: Goodfellow M and Minnikin DE (ed). \u003cem\u003eChemical Methods in Bacterial Systematics\u003c/em\u003e. Academic Press, London, pp 267\u0026ndash;287.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDe Ley J, Segers P, Gillis M. (1978) Intra- and Intergeneric Similarities of Chromobacterium and Janthinobacterium Ribosomal Ribonucleic Acid Cistrons. \u003cem\u003eInt. J. Syst. Bacteriol.\u003c/em\u003e 1978; 28:154\u0026ndash;168. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/00207713-28-2-154\u003c/span\u003e\u003cspan address=\"10.1099/00207713-28-2-154\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDong L, Ming H, Zhou EM, et al (2015) \u003cem\u003eCrenobacter luteus\u003c/em\u003e gen. nov., sp. nov., isolated from a hot spring. \u003cem\u003eInt J Syst Evol Microbiol\u003c/em\u003e 65:214\u0026ndash;219. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijs.0.060996-0\u003c/span\u003e\u003cspan address=\"10.1099/ijs.0.060996-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eFelsenstein J (1981) Evolutionary trees from DNA sequences: a maximum likelihood approach. \u003cem\u003eJ Mol Evol\u003c/em\u003e 17:368\u0026ndash;376. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF01734359\u003c/span\u003e\u003cspan address=\"10.1007/BF01734359\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eFelsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783\u0026ndash;791. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/j.1558-5646.1985.tb00420.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1558-5646.1985.tb00420.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eFitch WM (1971) Toward defining the course of evolution: Minimum change for a specific tree topology. \u003cem\u003eSyst Zool\u003c/em\u003e 20:406\u0026ndash;416. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/sysbio/20.4.406\u003c/span\u003e\u003cspan address=\"10.1093/sysbio/20.4.406\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGerhardt P, Murray RGE, Wood WA, Krieg NR (1994) Methods for General and Molecular Bacteriology. Amer Soc Microbiol.Washington DC.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eGrimes DJ, Woese CR, MacDonell MT, Colwell RR (1997) Systematic study of the genus \u003cem\u003eVogesella\u003c/em\u003e gen. nov. and its type species, \u003cem\u003eVogesella indigofera\u003c/em\u003e comb. nov. \u003cem\u003eInt J Syst Bacteriol\u003c/em\u003e 47:19\u0026ndash;27. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/00207713-47-1-19\u003c/span\u003e\u003cspan address=\"10.1099/00207713-47-1-19\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eJyoti V, Narayan KD, Das SK (2010) \u003cem\u003eGulbenkiania indica\u003c/em\u003e sp. nov., isolated from a sulfur spring. \u003cem\u003eInt J Syst Evol Microbiol\u003c/em\u003e 60:1052\u0026ndash;1055. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijs.0.014035-0\u003c/span\u003e\u003cspan address=\"10.1099/ijs.0.014035-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKai B, Shaw S, Kloosterman AM, Charlop-Powers Z, Weber T (2021) Antismash 6.0: improving cluster detection and comparison capabilities. \u003cem\u003eNucleic Acids Res.\u003c/em\u003e 49(W1): W29-W35. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/nar/gkab335\u003c/span\u003e\u003cspan address=\"10.1093/nar/gkab335\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKimura M (1980) A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. \u003cem\u003eJ Mol Evol\u003c/em\u003e16:111\u0026ndash;120. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/BF01731581\u003c/span\u003e\u003cspan address=\"10.1007/BF01731581\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKomagata K, Suzuki K (1987) Lipid and cell-wall analysis in bacterial systematic. \u003cem\u003eMethods Microbiol\u003c/em\u003e 19:161\u0026ndash;207. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/S0580-9517(08)70410-0\u003c/span\u003e\u003cspan address=\"10.1016/S0580-9517(08)70410-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKwon SW, Kim BY, Kim WG, et al (2008) \u003cem\u003ePaludibacterium yongneupense\u003c/em\u003e gen. nov., sp. nov., isolated from a wetland, Yongneup, in Korea. \u003cem\u003eInt J Syst Evol Microbiol\u003c/em\u003e 58:190\u0026ndash;194. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijs.0.64831-0\u003c/span\u003e\u003cspan address=\"10.1099/ijs.0.64831-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLane DJ, Goodfellow M (1991) 16S/23S rRNA sequencing. In: E. Stackebrandt. Chichester (ed). Nucleic Acid Techniques in Bacterial Systematics, Wiley, New York, pp 115\u0026ndash;175.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLarkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA \u003cem\u003eet al\u003c/em\u003e (2007) Clustal W and Clustal X version 2.0. \u003cem\u003eBioinformatics\u003c/em\u003e 23:2947\u0026ndash;2948. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/bioinformatics/btm404\u003c/span\u003e\u003cspan address=\"10.1093/bioinformatics/btm404\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLau HT, Faryna J, Triplett EW (2006) \u003cem\u003eAquitalea magnusonii\u003c/em\u003e gen. nov., sp. nov., a novel Gram-negative bacterium isolated from a humic lake. \u003cem\u003eInt J Syst Evol Microbiol\u003c/em\u003e56:867\u0026ndash;871. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijs.0.64089-0\u003c/span\u003e\u003cspan address=\"10.1099/ijs.0.64089-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLin MC, Chou JH, Arun AB, Young CC, Chen WM (2008) \u003cem\u003ePseudogulbenkiania subflava\u003c/em\u003e gen. nov., sp. nov., isolated from a cold spring. \u003cem\u003eInt J Syst Evol Microbiol\u003c/em\u003e 58:2384\u0026ndash;2388. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijs.0.65755-0\u003c/span\u003e\u003cspan address=\"10.1099/ijs.0.65755-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLiu ZX, Phurbu D, Liu HC, Zhou YG, Li AH (2020) \u003cem\u003eCraterilacuibacter sinensis\u003c/em\u003e gen. nov. sp. nov., isolated from a crater lake in China. \u003cem\u003eInt J Syst Evol Microbiol\u003c/em\u003e 70:4831\u0026ndash;4837. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijsem.0.004353\u003c/span\u003e\u003cspan address=\"10.1099/ijsem.0.004353\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLuo C, Rodriguez-R LM, Konstantinidis KT (2014) MyTaxa: an advanced taxonomic classifier for genomic and metagenomic sequences. \u003cem\u003eNucleic Acids Res\u003c/em\u003e 42:e73. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/nar/gku169\u003c/span\u003e\u003cspan address=\"10.1093/nar/gku169\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMeier-Kolthoff JP, G\u0026ouml;ker M. TYGS is an automated high-throughput platform for state-of-the-art genome-based taxonomy. \u003cem\u003eNat Commun.\u003c/em\u003e 2019;10:2182. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1038/s41467-019-10210-3\u003c/span\u003e\u003cspan address=\"10.1038/s41467-019-10210-3\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMiller LT (1982) Single derivatization method for routine analysis of bacterial whole-cell fatty acid methyl esters, including hydroxy acids. \u003cem\u003eJ Clin Microbiol\u003c/em\u003e 16:584\u0026ndash;586. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1128/jcm.16.3.584-586.1982\u003c/span\u003e\u003cspan address=\"10.1128/jcm.16.3.584-586.1982\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMinh BQ, Schmidt HA, Chernomor O, \u003cem\u003eet al\u003c/em\u003e (2020) IQ-TREE 2: New Models and Efficient Methods for Phylogenetic Inference in the Genomic Era. \u003cem\u003eMol Biol Evol.\u003c/em\u003e 37:2461. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/molbev/msaa131\u003c/span\u003e\u003cspan address=\"10.1093/molbev/msaa131\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMinnikin DE, Bolton RC, Hartmann S \u003cem\u003eet al.\u003c/em\u003e (1993) An integrated procedure for the direct detection of characteristic lipids in tuberculosis patients. \u003cem\u003eAnn Soc Belg Med Trop\u003c/em\u003e 73 Suppl 1:13\u0026ndash;24.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRichter M, Rossell\u0026oacute;-M\u0026oacute;ra R. (2009) Shifting the genomic gold standard for the prokaryotic species definition. \u003cem\u003eProc Natl Acad Sci\u003c/em\u003e USA 106:19126\u0026ndash;19131. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1073/pnas.0906412106\u003c/span\u003e\u003cspan address=\"10.1073/pnas.0906412106\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSaitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. \u003cem\u003eMol Biol Evol\u003c/em\u003e 4:406\u0026ndash;425. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/oxfordjournals.molbev.a040454\u003c/span\u003e\u003cspan address=\"10.1093/oxfordjournals.molbev.a040454\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSasser M (1990) Identification of bacteria by gas chromatography of cellular fatty acids, MIDI Technical Note 101. NewYork: MIDI Inc\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSedl\u0026aacute;ček I, Kwon SW, Švec P, Mašlanŏv\u0026aacute; I, K\u0026yacute;rov\u0026aacute; K, \u003cem\u003eet al\u003c/em\u003e (2016) \u003cem\u003eAquitalea pelogenes\u003c/em\u003e sp. nov., isolated from mineral peloid. \u003cem\u003eInt J Syst Evol Microbiol.\u003c/em\u003e 66:962\u0026ndash;967. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijsem.0.000819\u003c/span\u003e\u003cspan address=\"10.1099/ijsem.0.000819\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSheu SY, Chen ZH, Young CC, Chen WM (2014) \u003cem\u003ePaludibacterium paludis\u003c/em\u003e sp. nov., isolated from a marsh. \u003cem\u003eInt J Syst Evol Microbiol\u003c/em\u003e 64:2497\u0026ndash;2502. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijs.0.060954-0\u003c/span\u003e\u003cspan address=\"10.1099/ijs.0.060954-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSubhash Y, Tushar L, Sasikala C, Ramana CV (2013) \u003cem\u003eVogesella alkaliphila\u003c/em\u003e sp. nov., isolated from an alkaline soil, and emended description of the genus \u003cem\u003eVogesella\u003c/em\u003e. \u003cem\u003eInt J Syst Evol Microbiol.\u003c/em\u003e 63:2338\u0026ndash;2343. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijs.0.046300-0\u003c/span\u003e\u003cspan address=\"10.1099/ijs.0.046300-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eTamura K, Peterson D, Peterson N, Stecher G, Nei M \u003cem\u003eet al\u003c/em\u003e (2011) MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. \u003cem\u003eMolecular Biology and Evolution\u003c/em\u003e 10:2731\u0026ndash;2739. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1093/molbev/msr121\u003c/span\u003e\u003cspan address=\"10.1093/molbev/msr121\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eVaz-Moreira I, Nobre MF, Nunes OC, Manaia CM (2007) \u003cem\u003eGulbenkiania mobilis\u003c/em\u003e gen. nov., sp. nov., isolated from treated municipal wastewater. \u003cem\u003eInt J Syst Evol Microbiol\u003c/em\u003e 57:1108\u0026ndash;1112. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijs.0.64726-0\u003c/span\u003e\u003cspan address=\"10.1099/ijs.0.64726-0\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWiney M, Meehl JB, O'Toole ET, Giddings TH Jr. (2014) Conventional transmission electron microscopy. \u003cem\u003eMol Biol Cell\u003c/em\u003e 25:319\u0026ndash;23. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1091/mbc.E12-12-0863\u003c/span\u003e\u003cspan address=\"10.1091/mbc.E12-12-0863\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWu C, Lu X, Qin M \u003cem\u003eet al\u003c/em\u003e (1989) Analysis of menaquinone compound in microbial cells by HPLC. \u003cem\u003eMicrobiology [English translation of Microbiology (Beijing)]\u003c/em\u003e 16:176\u0026ndash;178.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eYoon SH, Ha SM, Kwon S \u003cem\u003eet al.\u003c/em\u003e (2017) Introducing EzBioCloud: a taxonomically united database of 16S rRNA gene sequences and whole-genome assemblies. \u003cem\u003eInt J Syst Evol Microbiol\u003c/em\u003e 67:1613\u0026ndash;1617. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1099/ijsem.0.001755\u003c/span\u003e\u003cspan address=\"10.1099/ijsem.0.001755\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\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":"antonie-van-leeuwenhoek","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"anto","sideBox":"Learn more about [Antonie van Leeuwenhoek](https://www.springer.com/journal/10482)","snPcode":"10482","submissionUrl":"https://submission.nature.com/new-submission/10482/3","title":"Antonie van Leeuwenhoek","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"crater lake, Craterilacuimicrobium, Da Hinggan mountain, Neisseriales","lastPublishedDoi":"10.21203/rs.3.rs-7595832/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7595832/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eStrain TC5R-5\u003csup\u003eT\u003c/sup\u003e, a white rod-shaped bacterium, was isolated from Aershan Tianchi crater lake in northern China. It is Gram-stain-negative, catalase- and oxidase- positive. Optimal growth occurs at 20\u0026ndash;30\u0026deg;C, pH of 6.0\u0026ndash;7.0 and in the presence of 0-0.5% (w/v) NaCl. Based on 16S rRNA gene sequences analysis, strain TC5R-5\u003csup\u003eT\u003c/sup\u003e exhibits the highest similarity (94.3%) to \u003cem\u003eChromobacterium alkanivorans\u003c/em\u003e IITR71\u003csup\u003eT\u003c/sup\u003e. The genomic DNA G\u0026thinsp;+\u0026thinsp;C content of strain TC5R-5\u003csup\u003eT\u003c/sup\u003e is 51.4% according to the genome sequence. The predominant cellular fatty acids are summed feature 3 (comprising C\u003csub\u003e16:1\u003c/sub\u003e \u003cem\u003eω\u003c/em\u003e7\u003cem\u003ec\u003c/em\u003e /C\u003csub\u003e16:1\u003c/sub\u003e \u003cem\u003eω\u003c/em\u003e6\u003cem\u003ec\u003c/em\u003e) and C\u003csub\u003e16:0\u003c/sub\u003e. The predominant quinone is ubiquinone Q-8, with Q-7 present in moderate amounts. The polar lipid profile comprises phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), an unidentified aminophospholipid (APL), and an unidentified phospholipid (PL). Through polyphasic taxonomy and molecular phylogenetic analysis, strain TC5R-5\u003csup\u003eT\u003c/sup\u003e is proposed to represent a novel species of a new genus of the family \u003cem\u003eChromobacteriaceae\u003c/em\u003e within the order \u003cem\u003eNeisseriales\u003c/em\u003e, for which the name \u003cem\u003eCraterilacuimicrobium aquaticum\u003c/em\u003e gen. nov. sp. nov. is proposed. The type strain is TC5R-5\u003csup\u003eT\u003c/sup\u003e (=\u0026thinsp;CGMCC 1.17058\u003csup\u003eT\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;KCTC 72736\u003csup\u003eT\u003c/sup\u003e).\u003c/p\u003e","manuscriptTitle":"Craterilacuimicrobium aquaticum gen. nov. sp. nov., a novel member of the order Neisseriales isolated from crater lake","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-10-18 06:14:07","doi":"10.21203/rs.3.rs-7595832/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-02-05T05:08:15+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-26T18:16:33+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"103054189930880707451816252604788654543","date":"2025-11-20T10:24:32+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-13T09:16:47+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"327234333595436244936428698205299894542","date":"2025-10-07T10:35:49+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-10-06T09:29:03+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-12T10:13:53+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-09-12T04:51:59+00:00","index":"","fulltext":""},{"type":"submitted","content":"Antonie van Leeuwenhoek","date":"2025-09-12T02:43:15+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"antonie-van-leeuwenhoek","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"anto","sideBox":"Learn more about [Antonie van Leeuwenhoek](https://www.springer.com/journal/10482)","snPcode":"10482","submissionUrl":"https://submission.nature.com/new-submission/10482/3","title":"Antonie van Leeuwenhoek","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"0eb23407-3b2f-46a9-bc74-112b5f6501bd","owner":[],"postedDate":"October 18th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-03-03T04:24:23+00:00","versionOfRecord":[],"versionCreatedAt":"2025-10-18 06:14:07","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7595832","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7595832","identity":"rs-7595832","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}