A Proteomic Study to Elucidate Molecular Relationships Between Iron, Oxidative Stress and Polyphosphate in  Streptomyces coelicolor A3(2)

A Proteomic Study to Elucidate Molecular Relationships Between Iron, Oxidative Stress and Polyphosphate in Streptomyces coelicolor A3(2) | 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 A Proteomic Study to Elucidate Molecular Relationships Between Iron, Oxidative Stress and Polyphosphate in Streptomyces coelicolor A3(2) Şerif Yılmaz, Filiz Yeşilırmak, Sedef Tunca This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4107881/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 19 Jul, 2024 Read the published version in Biologia → Version 1 posted 5 You are reading this latest preprint version Abstract Polyphosphate (polyP) is an important energy and phosphate storage polymer in all organisms. Deletion of the polyP synthesising enzyme, polyP kinase (PPK), resulted in an antibiotic overproducing phenotype in Streptomyces . However, the industrial use of overproducing Streptomyces strains without PPK activity (∆ ppk ) is hampered by their increased sensitivity to oxidative stress. Iron plays a key role in the bacterial response to oxidative stress, and it is also an essential element for various processes in the cell. Conversely, polyP can sequester iron, reducing its bioavailability. This study aimed to elucidate the metabolic relationship between oxidative stress, iron, and polyP metabolisms in Streptomyces coelicolor as an example of the communication of cellular regulatory signalling networks. Comparative proteomic analyses were performed on three biological replicates of wild-type and ∆ ppk strains grown in iron-containing and iron-free media. Independent of iron, the results show that the absence of polyP significantly alters the total proteome, revealing the importance of this polymer in maintaining cellular metabolism. The mutant strain was found to have difficulties coping with the iron even in the nutrient-rich medium. Compared to the wild type in the iron-free medium, a general abundance of proteins related to energy metabolism, and protein folding was observed in ∆ ppk . In the presence of iron, the expression of the proteins involved in translation, phosphate metabolism and the antioxidant system was increased in the mutant strain compared to the wild type. To our knowledge, this is the first study to clarify the relationship between iron, oxidative stress, and polyphosphate. Oxidative stress iron polyphosphate 2D gel electrophoresis proteomic Streptomyces coelicolor A3(2) Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Streptomyces species are well known for their ability to synthesize valuable secondary metabolites. To develop Streptomyces -based production hosts with optimized metabolic backgrounds, we need to have a holistic view of the molecular mechanisms that orchestrate secondary metabolism. The "cause and effect" view must therefore be replaced by the integrative study of cellular regulatory signaling networks. Phosphate (Pi) availability and the oxidative stress response are two factors able to modulate Streptomyces secondary metabolism(Esnault et al., 2017). Pi limitation is a trigger of secondary metabolite production and disruption of homeostasis of intracellular reactive oxygen species (ROS) causes a redirection of metabolic fluxes (Beites et al., 2011) . Pi metabolism in Streptomyces is regulated by the two-component system PhoRP (Liu et. al., 2013). Under Pi starvation conditions, the membrane sensor kinase PhoR phosphorylates the response regulator PhoP that controls the expression of more than 40 genes that comprise the PHO regulon and are required for phosphate scavenging and assimilation. However, while maintaining Pi homeostasis, PhoP also mediates the transient shutdown of central metabolic pathways, secondary metabolism, and morphological differentiation (Allenby et al., 2012) . Over the last decade, experimental data suggests a crosstalk between Pi and ROS homeostasis. In Pi limiting conditions, an indirect PhoP-dependent transcription of oxidative stress related genes (notably ahpC coding for the alkyl hydroperoxide reductase, an H 2 O 2 detoxifying enzyme) is observed (Rodriguez-Garcia et. al., 2007) together with enhanced sensitivity to oxidative stress (Ghorbel et. al., 2006). On the other hand, the S. natalensis Δ ahpC strain that presented high intracellular ROS levels, displayed a delayed Pi uptake and an up-regulation of the PHO regulon, namely polyphosphate kinase gene ( ppk ) (Beites et al., 2011). Polyphosphate kinase enzyme (PPK) is responsible for synthesis of an important energy and phosphate storage polymer, polyphosphate (polyP), and its transcription is regulated by PhoP. Deletion of ppk in S. coelicolor resulted in an actinorhodin overproducing phenotype (Yalim Camci et al., 2012). However, the industrial exploitation of overproducing Streptomyces strains without PPK activity is hampered by their increased sensitivity to oxidative stress (Yalim Camci et al., 2012; Ghorbel et. al., 2006). Worth noting that both Pi metabolism and the oxidative stress response are two physiological processes able to modulate the availability of intracellular iron. Iron is an essential element for bacterial vegetative growth and secondary metabolism (Weinberg, 1990) and it plays a key role in the bacterial oxidative stress response either as a cofactor in ROS-sensing proteins or by the generation of the highly toxic hydroxyl radical via the Fenton reaction (Imlay, 2013). Although cells have ROS-degrading enzymes such as superoxide dismutases, catalases, alkylhydroperoxidases, etc. (Cornelis et al., 2011), polyP also prevents oxidative stress by inhibiting Fenton reactions via chelating iron (Gray & Jakob, 2015). At the same time, polyP mediates the appropriate refolding of the damaged proteins in oxidative stress conditions (Gray et al., 2014). Independent from polyP, it has recently been shown that iron is stored in gram-positive Clostridioides difficile in the form of "iron-phosphate granules" (Pi et. al., 2023). In recent years, significant progress has been made in the study of the molecular mechanisms controlling secondary metabolism in Streptomyces . However, there is still insufficient information on the interactions between different regulatory mechanisms. This study aims to elucidate the metabolic relationship between oxidative stress, iron, and Pi metabolism of S. coelicolor by comparative proteomic analyses of the wild-type and Δ ppk strains grown in iron-containing and iron-free media. Material and Methods Media and growth conditions S. coelicolor A3(2) and Δ ppk strains were grown in TSB and R2YE media at 30°C (Kieser et al., 2000). For protein isolation, seed cultures were prepared by growing the bacteria in TSB. After centrifugation at 1000×g for 15 min. bacterial pellets were weighed and equal amounts from each strain were inoculated into R2YE broth and were grown for 48 hours at 30°C and 250 rpm. The presence of 0.0025 mM FeCl 3 in the R2YE medium was ignored and used as the iron-free medium. R2YE medium was supplemented with 100 mM FeCl 3 to obtain an iron-containing medium. All the flasks were washed with 8M HCI to get rid of iron contamination, rinsed, and autoclaved. Protein Isolation Bacterial cells, grown for 48 hours at 30°C, 250 rpm, were collected by centrifugation at 1000×g for 15 min. and the proteins were isolated according to Faurobert et al. (2007). Each pellet was dissolved in 2 mL extraction buffer (500 mM Tris-HCl, 50 mM EDTA, 700 mM sucrose, 100 mM KCl, pH 8.0, 1X protease inhibitor cocktail) and the cells were lysed by sonication (15 s on/30 s off, 50% of amplitude) for 5 minutes. After incubation on ice for 10 minutes, an equal volume of Tris-buffered phenol (pH8.0, Sigma) was added and the solution was shaken at room temperature for 10 min. Then the sample was centrifuged at 3200×g for 10 min and the supernatant was carefully transferred to the new tube. After adding 3 mL extraction buffer to the sample, it was vortexed and shaken for 3 min at room temperature. Then the sample was centrifuged at 3200×g for 10 min. and 4 volumes of precipitation buffer (0.1 M ammonium acetate dissolved in methanol) was added onto the supernatant and incubated at -20 ° C for at least 1 day. Then the proteins were precipitated by centrifugation at 3200×g for 10 min. and they were treated 3 times with cold precipitation buffer and once with cold acetone. Isolated proteins were visualized on SDS-PAGE (Laemmli, 1970), and their concentrations were calculated by the Bradford method (Bradford, 1976) using bovine serum albumin as a control. Protein Separation by 2D-PAGE 400 µg of protein sample was mixed with 3 ml rehydration buffer (7M urea, 2 M thiourea, 4% w/v CHAPS, 0.46% w/v DTT) and loaded on IPG strips (17 cm, pH 3–10 nonlinear gradient, Bio-Rad, USA). IPG strips were first passively rehydrated for 2 hours and then actively rehydrated with 50 V current for 16 hours. Isoelectric focusing was carried out at 20°C on the IPGphor to be a total of 6000V. The strips were incubated in equilibration solution (6 M urea, 30% w/v glycerol, 2% w/v SDS, 50 mM Tris, pH 8.8) containing 1% w/v DTT, followed by 15 minutes incubation in 4% w/v iodoacetamide equilibration solution. After isoelectric focusing, the second dimension was performed in 12% polyacrylamide gels. The gels were stained with colloidal Coomassie brilliant blue solution and the gel image was transferred to a computer using a digital imaging system (VersaDoc MP 4000, Bio-Rad). Image analysis of 2D Gels For spot detection “Spot colocalisation (ComDet) plug-in” and for quantification “pixel intensity” programs in ImageJ (Fiji), were used. ImageJ calculates the average pixel value in areas with grayscale values ranging from 0 (black) to 255 (white). Three biological replicates of each sample (WT and Δ ppk ), grown in iron-free and iron supplemented media, resulted in 12 SDS PAGE gels. The result for each spot is expressed as the ratio between Δ ppk and WT spot intensity; and between iron-supplemented and iron-free medium for Δ ppk and WT. Statistical Analysis of Variations in Protein Abundance Normalisation is used as a technique used to adjust data from different samples in order to eliminate unwanted variability and improve comparability between datasets (Carvalho et al., 2024). In this study, we performed the quantile normalisation and minimum-maximum scaling normalisations performed by Zhao et al. (2020) on three replicates obtained in R2YE and iron supplemented R2YE for both Δ ppk and WT. Statistical analysis according to Augillian et al. (2020) was used. After data normalisation, fold change values were calculated, and the F -test was used for analysis of variation. The F -test was evaluated at a significance level of p < 0.05. If the F -test is p < 0.05, the variation is unequal (heteroskedastic), otherwise the variation is equal (homoskedastic). Independent two-sample t -test was performed according to these two variation conditions. The results of the two-sample t -test were evaluated at a significance level of p < 0.05 to assess the expression differences of the proteins. Protein identification by matrix-assisted laser desorption/ionization tandem time of flight (MALDITOF-TOF) Spots were cut out of the gel and washed two times with a solution of 50% (v/v) methanol and 5% acetic acid until they became colorless. Then they were dehydrated with acetonitrile (ACN), treated with a solution of 10 mM DTT in 100 mM NH 4 HCO 3 for 30 minutes at room temperature, and subsequently alkylated with a solution of 100 mM iodoacetamide in 100 mM NH 4 HCO 3 for 30 minutes in dark. Following further dehydration with ACN and rehydration with NH 4 HCO 3 , the gel fragments were digested with 30 µL of trypsin solution (20 ng/µL prepared in 100 mM NH 4 HCO 3 ) and incubated at 37°C overnight. The peptides were then extracted twice by using a solution of 5% formic acid in 50% ACN and then desalted with ZipTip. Mass spectrometry (MS) analysis was carried out by using a MALDI-TOF-TOF instrument (Bruker Autoflex III Smartbeam, USA), and the obtained spectra were processed and analyzed with the BioTools software (Bruker Daltonics, USA). Database searching was conducted separately using an in-house MASCOT server (Matrix Science, London, UK). Results The effect of iron on the proteome profile of S. coelicolor A3(2) (WT) and Δ ppk mutant strains was studied. The cells were grown in liquid R2YE and R2YE supplemented with 100 mM FeCl 3 . Proteins were isolated from 48-hour grown cells to make a comparison before toxic levels of ROS accumulate in the presence of iron. Isolated proteins were run on 2D gels and a total of 12 SDS PAGE gels of WT and Δ ppk strains of 3 biological replicates were obtained. A representation of the protein gels for the two strains is shown in Fig. 1 . Protein identification was performed by protein fingerprinting, and out of 78 spots, 66 were successfully identified by using the fully sequenced and annotated genome of S. coelicolor (Bentley et al., 2002), while 12 remained unidentified. Of the 66 spots identified, 37 represented distinct proteins. These proteins were classified into 6 categories (energy metabolism, phosphate metabolism, oxidative stress metabolism, translation, chaperone systems, others) and protein interaction map was generated using the STRING database (Fig. 2 ). It was observed that in the R2YE medium, the expression of 16 proteins was increased, 3 were decreased and 18 were not considerably altered in the Δ ppk strain compared to the WT strain (Fig. 3 , Table). In the iron-supplemented medium, the expression of 12 proteins increased, 6 decreased and 19 showed no significant change in the mutant strain compared to the WT strain (Fig. 3 ; Table). According to the protein profile of the Δ ppk strain grown in iron-free and iron-supplemented medium, the expression of 6 proteins was found to increase, 18 of them decreased and 13 showed no significat change in the presence of iron (Table). In the WT strain grown in iron-supplemented medium compared to iron-free medium, the expression of 6 proteins increased, 11 decreased and 20 proteins were not significantly changed (Table). Energy Metabolism Compared to the WT strain, dihydrolipoamide dehydrogenase (spot 4), 2-oxoglutarate dehydrogenase (spot 12), and dihydrolipoamide succinyltransferase (spot 27) proteins were abundant in the Δ ppk strain in R2YE medium (Table, Fig. 4 ). These three enzymes form a highly conserved “2-oxoglutarate dehydrogenase complex” which plays an important role in cellular energy metabolism by catalysing the conversion of 2-oxoglutarate to succinyl-CoA and NADH (+ H + ) in the tricarboxylic acid cycle (Chen et al., 2019). This multi-enzyme system also protects against oxidative stress through interactions with alkyl hydroperoxidases (AhpC and AhpD) (Su et al., 2019). Phosphoenolpyruvate carboxykinase (PEPCK) (spot 13), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (spot 8) proteins were also found to be increased in Δ ppk strain in R2YE medium (Table, Fig. 4 ). PEPCK is a crucial enzyme involved in regulating the carbon flow within the central metabolism. It captures CO 2 and incorporates it into PEP to generate oxaloacetate or catalyze the reverse reaction (Chiba et al., 2015). GAPDH is responsible for catalyzing the sixth step of glycolysis and also contributes to gluconeogenesis. Transketolase (spot 15) was another enzyme whose expression was elevated in the Δ ppk strain compared to WT in the absence of iron. Its function is to link glycolysis and the pentose phosphate pathway (Vimala & Harinarayanan, 2016) (Table, Fig. 4 ). Upon the addition of iron to the medium, the mutant cell showed a decrease in energy metabolism when compared to the iron-depleted conditions. This suggests that the cell is attempting to stabilize the amount of ROS that has already formed due to Fenton reactions, as energy metabolism is one of the primary sources of ROS in cells. In the iron-supplemented medium, only glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein was found to be increased in Δ ppk compared to the WT strain. Similarly, the presence of iron did not significantly affect the protein profile of the WT strain. However, dihydrolipoamide succinyltransferase (spot 27) was found to be increased, while fructose bisphosphate aldolase (spot 32) was found to be decreased. Phosphate Metabolism Compared to WT, PhoP (spot 17), PhoX (spot 23) and PhoU (spot 26) proteins were found to be abundant in the Δ ppk strain in iron supplemented medium. In the absence of iron, only PhoX protein was found to be decreased (Table, Fig. 4 ). PhoP controls more than 40 genes involved in phosphate scavenging and assimilation. PhoU is the modulator of phosphate-specific transport system and PhoX is an alkaline phosphatase that functions under Pi-limiting conditions (Martín & Liras, 2021). The presence of iron caused a slight decrease in the expression of PhoP and PhoX in WT. Oxidative stress Myo-inositol-phosphate synthase, superoxide dismutase (SOD) and catalase play essential roles in oxidative stress resistance. SOD and catalase enzymes catalyze the conversion of reactive oxygen species into harmless forms (Kanth et al., 2011). Myo-inositol-phosphate synthase the conversion of D-glucose 6-phosphate to 1L-myo-inositol 1-phosphate (Geiger & Jin, 2006), which is the precursor of an antioxidant agent mycothiol (Chen et al., 2019). Compared to WT, SOD (spot 20) and myo-inositol phosphate synthase (spot 7) were found to be more abundant and catalase (spot 31) was less abundant in the Δ ppk strain in the presence of iron. Presence of iron caused a decrease in the expression of both SOD and myo-inositol phosphate synthase in the mutant strain, compared to that grew in the absence of iron. In the WT strain, SOD was found to be less abundant and catalase was found to be more abundant in the presence of iron compared to the WT strain in the R2YE (Table, Fig. 4 ). Translation Compared to the WT strain, ribosomal proteins S1 (spot 2) and S10 (spot 39) were found to be abundant, while ribosomal protein L17 (spot 24) and Elongation factor Tu-1 (spot 37) were found to be decreased in Δ ppk strain in R2YE medium. In the iron-supplemented medium, ribosomal proteins S10 (spot 39), S16 (spot 34), L10 (spot 35), L17 ribosomal proteins (spot 24) were found to be up-regulated together with proline tRNA ligase (spot 25) in the Δ ppk strain, compared to WT. The increase in the expression level of ribosomal protein S16 was very pronounced (log2 fold change 4,3). The presence of iron caused an increase in the expression of ribosomal protein S1; and a decrease in L17 and S10 in WT (Table, Fig. 4 ). Δ ppk grown in iron, expressed more L17, L10, S16, Elongation factor Tu-1 and proline-tRNA ligase than those grown in iron-free medium. Proline-tRNA ligase catalyses the addition of proline to tRNA and elongation factor Tu-1 brings the aminoacyl tRNAs to the aminoacyl region of the ribosome (Rodnina, 2016). Protein Folding and Proteases Among the major chaperone proteins, the trigger factor (TF) (spot 1), GroES (spot 11) and GroEL (spot 33) proteins, the Clp family protease (spot 14) and tricorn protease (spot 29) proteins were abundant in Δ ppk compared to WT in R2YE medium. It is known that differential expression of Clp protease genes affects the differentiation process in Streptomyces (Bellier & Mazodier, 2004). On the other hand, the tricorn protease is involved in the recycling of proteins within the cell (Tamura et al., 2001). In iron supplemented medium, GroES was found to be increased and Clp family protease and tricorn protease were decreased in Δ ppk strain compared to WT. Similarly, except for GroES, TF, GroEL, the Clp protease and tricorn protease were less abundant in the Δ ppk strain in the presence of iron compared to iron-free conditions. Clp family protease and tricorn protease were increased, GroEL and GroES on the other hand was decreased in WT in the iron supplemented medium (Table, Fig. 4 ). Other Proteins N-acetylneuraminate synthase (spot 9) and N-acylneuraminate cytidylyltransferase (spot 3) proteins were abundant in Δ ppk strain in R2YE medium compared to WT. Both enzymes participate in aminosugar metabolism, specifically related with cell wall synthesis (Bravo et al., 2001). Ribbon-helix-helix protein (spot 19), which is a transcription factor that plays an important role in the regulation of metal uptake, amino acid biosynthesis, cell division, etc. (Schreiter & Drennan, 2007), was found to be also slightly increased in Δ ppk compared to WT in R2YE medium. In the presence of iron, the expression of this protein, as well as N-acetylneuraminate synthase, N-acylneuraminate cytidylyltransferase, and glutamate dehydrogenase (spot 28), were found to be decreased in the Δ ppk strain compared to iron-free conditions. Glutamate dehydrogenase plays a major role in amino acid metabolism by catalysing the reversible process of oxidative deamination, converting glutamate to α-ketoglutarate (Smith et al., 2019). Compared to the WT strain, glutamate dehydrogenase and RNA polymerase subunit beta' (spot 30) were found to be reduced in Δ ppk in iron supplemented R2YE. RNA polymerase subunit beta' is one of the major subunits of the RNA polymerase holoenzyme (Sutherland & Murakami, 2018). When all the results were analysed together, it was observed that compared to the wild type strain, the levels of proteins involved in energy metabolism and the chaperone system were increased in the mutant strain in iron-free medium, whereas the levels of SOD and mycothiol, which are elements of the antioxidant system, were increased in iron-containing medium, together with proteins involved in phosphate metabolism and translation. When comparing the protein profiles of the mutant strain in the presence and absence of iron, it is evident that the strain has difficulty coping with the iron in the absence of polyphosphate. In the presence of iron, it only increases the expression of translational proteins and PhoX, while decreasing the expression of most of the proteins, identified in this study that are involved in other metabolisms. While the presence of iron did not significantly alter the total proteome of the wild-type strain, it did cause a decrease in the expression of PhoP, PhoX, Fructose-bisphosphate aldolase, SOD, ribosomal proteins L17 and S10. The concentration of iron used in this study did not cause toxicity in the wild-type strain. On the contrary, it caused a decrease in the expression of UspA, a putative stress protein. Table. Differentially expressed proteins ( ↑ : increase, ↓: decrease, ─: no significant differences) Spot No Protein Name Δppk/ WT Fold Change log2 ΔppkFe/ WTFe Fold Change log2 ΔppkFe/ Δppk Fold Change log2 WTFe/ WT Fold Change log2 Gene UniProt Mw pI PMF MSMS Energy Metabolism 4 Dihydrolipoamide dehydrogenase ↑ 0,8* ─ -0,3 ↓ -1,4* ─ -0,3 SCO2180 Q9S2Q6 51536 5,92 17 1 8 Glyceraldehyde-3-phosphate dehydrogenase ↑ 0,6* ↑ 0,4* ─ -0,4 ─ -0,1 SCO7511 Q93J08 35003 5,41 11 2 12 Putative 2-oxoglutarate dehydrogenase ↑ 1,2* ─ -0,9 ↓ -1,6* ─ 0,5 SCO5281 Q9FBR4 139292 5,91 20 2 13 Phosphoenolpyruvate carboxylase ↑ 1,6* ─ 0,4 ↓ -1,1* ─ 0,1 SCO3127 Q9RNU9 101349 5,48 32 2 15 Transketolase ↑ 0,5* ─ 0,1 ─ -0,5 ─ -0,1 SCO1935 Q9XAC1 75085 5,19 13 3 27 Dihydrolipoamide succinyltransferase ↑ 0,7* ─ -0,4 ↓ -0,6* ↑ 0,5* SCO2181 Q9S2Q5 59000 4,57 9 3 32 Fructose-bisphosphate aldolase ─ 0,3 ─ 0,2 ↓ -0,8* ↓ -0,7* SCO3649 Q9X8R6 37018 5,42 11 2 Phosphate Metabolism 17 Putative response regulator (PhoP) ─ 0,3 ↑ 0,6* ─ -0,3 ↓ -0,6* SCO4230 Q9L0R1 24770 5,08 8 2 23 PhoX ↓ -1,0* ↑ 0,5* ↑ 1,4* ↓ -0,1* SCO3790 Q9F2J1 75179 6,21 11 2 26 Phosphate-specific transport system accessory protein PhoU ─ 1,0 ↑ 0,6* ↓ -0,2* ─ 0,2 SCO4228 Q8CJU3 25441 4,93 8 1 Oxidative stress 7 Myo-inositol-1-phosphate synthase ─ 0,1 ↑ 0,6* ↓ -0,6* ─ -1,1 SCO3899 Q9X8T5 39433 5,07 - 2 20 Superoxide dismutase [Fe-Zn] ─ 0,1 ↑ 0,5* ↓ -0,8* ↓ -1,2* SCO2633 O51917 23513 5,15 7 2 31 Catalase ─ -0,3 ↓ -1,3* ─ -0,9 ↑ 0,1* SCO0379 Q9RJK9 55139 5,84 21 * Translation 2 30S ribosomal protein S1 ↑ 0,5* ↓ -0,3* ↓ -0,6* ↑ 0,3* SCO1998 Q9S2K5 55001 4,54 16 5 6 30S ribosomal protein S2 ─ 0,2 ─ -0,2 ─ -0,6 ─ -0,3 SCO5624 O31212 33602 5,17 10 1 24 50S ribosomal protein L17 ↓ -1,3* ↑ 1,4* ↑ 1,4* ↓ -1,4* SCO4730 O86775 18120 9,42 6 2 25 Proline--tRNA ligase ─ 0,3 ↑ 1,6* ↑ 1,9* ─ 0,5 SCO5699 Q9KYR6 61443 4,98 9 - 34 30S ribosomal protein S16 ─ 0,6 ↑ 4,3* ↑ 1,1* ─ -2,7 SCO5591 O69879 15187 9,25 - 2 35 50S ribosomal protein L10 ─ 0,2 ↑ 1,0* ↑ 1,5* ─ 0,6 SCO4652 P41103 18635 8,78 11 2 37 Elongation factor Tu-1 ↓ -2,0* ─ -0,9 ↑ 0,9* ─ -0,2 SC04662 P40174 43811 5,01 11 3 39 30S ribosomal protein S10 ↑ 1,0* ↑ 1,2* ─ -0,2 ↓ -0,4* SCO4701 P66337 11571 9,3 10 2 Protein folding and Proteases 1 Trigger factor ↑ 1,5* ─ 0,0 ↓ -0,4* ─ 1,0 SCO2620 Q9F314 51137 4,31 13 2 11 Co-chaperonin GroES ↑ 0,8* ↑ 0,5* ─ -0,4 ↓ -0,1* SCO4761 P0A345 10940 4,65 5 - 14 Putative Clp-family ATP-binding protease ↑ 0,7* ↓ -1,0* ↓ -1,4* ↑ 0,3* SCO3373 Q9S6T8 92957 5,73 22 4 29 Tricorn protease homolog 1 ↑ 1,1* ↓ -2,1* ↓ -2,4* ↑ 0,9* SCO2549 Q9RDE2 115577 5,47 37 2 33 Chaperonin GroEL 2 ↑ 0,6* ─ 0,0 ↓ -0,6* ↓ -0,1* SCO4296 Q9KXU5 56795 4,79 15 3 Others 3 N-acylneuraminate cytidylyltransferase ↑ 2,0* ─ 0,2 ↓ -1,7* ─ 0,1 SCO4880 Q9AK46 44514 5,63 17 - 5 Phage Tail Protein ─ 0,1 ─ -0,2 ─ -0,6 ─ -0,3 SCO4252 Q9L0N9 16527 4,86 8 2 9 N-acetylneuraminate synthase ↑ 1,4* ─ 0,8 ↓ -0,8* ─ -0,2 SCO4880 Q9AK45 34716 5,29 18 5 10 UspA domain-containing protein ─ 0,1 ─ -0,1 ─ 0,1 ─ 0,3 SCO0200 Q9RI46 32237 5,9 12 3 16 Putative 3-oxoacyl-[acyl-carrier protein] reductase ─ -0,1 ─ 0,3 ─ -0,2 ↓ -0,6* SCO6282 Q93S07 27414 5,42 9 4 18 Conserved hypothetical membrane protein ─ -0,2 ─ -0,1 ↓ -0,6* ↓ -0,8* SCO3967 Q93J39 25827 5,07 9 2 19 Ribbon-helix-helix protein, CopG family ↑ 0,3* ─ -0,3 ↓ -1,1* ↓ -0,6* SCO5908 O54104 7146 - 6 - 28 Glutamate dehydrogenase ─ 0,1 ↓ -2,7* ↓ -2,7* ─ 0,1 SCO2999 Q8CJY0 183751 5,25 22 1 30 DNA-directed RNA polymerase subunit beta' ─ -0,7 ↓ -3,0* ─ -2,3 ─ 0,0 SCO4655 Q8CJT1 145199 6,44 10 - 36 Putative ABC transporter ATP-binding protein ─ 0,1 ─ 0,3 ─ 0,4 ─ 0,1 SCO3224 Q9S6T6 34069 6,22 11 2 40 UspA-Putative Stress Protein ─ -0,2 ─ 0,6 ─ -0,6 ↓ -1,3* SCO0167 Q9RIZ8 31222 5,66 6 2 × Fold change data showing that the comparison of protein data is significant ( p <0.05) Discussion Polyphosphate (polyP) is a conserved polymer that has important functions in every organism. One important function of polyP is to help cells resist various stresses. Streptomyces strains that are unable to produce polyP are known to have increased sensitivity to oxidative stress (Ghorbel et al., 2006; Yalim Camci et al., 2012; Le Maréchal et al., 2013). Iron is an important element in the cell, but it plays a role in the generation of oxidative stress by causing the formation of ROS compounds through Fenton reactions. In this study, comparative proteomic analyses of S. coelicolor ppk mutant (∆ ppk ) and wild-type (WT) strains were performed in the presence and absence of iron to understand the relationship between polyphosphate, oxidative stress, and iron metabolisms. It was found that in the absence of poly P, which is an important energy storage polymer, the ∆ ppk strain suffered from energy deprivation even in R2YE, a nutrient-rich medium that promotes Streptomyces growth. To overcome this energy deficit, the relevant pathways were activated in the mutant strain. Presence of iron did not caused significant changes in the abundance of energy metabolism proteins in Δ ppk compared to WT. However ∆ ppk strain reduced the expression of enzymes involved in energy metabolism in the presence of iron compared to the conditions without iron. Streptomyces are characterized by a high utilization of the Pentose Phosphate pathway and complex-I (NADH:ubiquinone oxidoreductase) respiratory enzymes, which under normal conditions produce more ROS (Lejeune et al., 2022). The TCA cycle is also an important source of ROS metabolites (Kohanski et al., 2007). It can be hypothesized that the mutant strain, which is already under oxidative stress, tries to reduce the amount of ROS by decreasing the expression of enzymes involved in energy metabolism. This result is not surprising since Streptomyces are known to change their metabolic networks under certain conditions. For example, Choi et al. (2022) showed that in S. coelicolor under oxygen-limited conditions, the presence of iron altered the cell metabolism and increased the NAD/NADH + ratio supporting anaerobic growth. The expression of proteins associated with phosphate metabolism such as PhoP, PhoU, and PhoX was found to be increased in the presence of iron in the ∆ ppk strain. It was clear that the mutant strain tried to increase the amount of free phosphate when there was iron in the environment. Phosphate metabolism in Streptomyces is regulated by a two-component system called PhoR/PhoP. In this system, PhoR functions as a sensor kinase while PhoP acts as a response regulator. Other than phosphate utilization and storage, PhoP protein has also been associated with oxidative stress resistance. In a study with S. coelicolor PhoP mutant, the expression of some genes related to resistance to oxidative stress was found to be reduced (Rodríguez-García et al., 2007). PhoP is also involved in the positive regulation of the ppk gene and indirectly of polyP, which chelates iron and prevents the formation of ROS by the Fenton reaction (Beaufay et al., 2020). PhoU is another protein that has been associated with oxidative stress. It was observed that H 2 O 2 sensitivity was increased in the mutant strain that could not produce PhoU protein (Ghorbel et al., 2006). Bacteria induce alkaline phosphatases when inorganic phosphate (Pi) is insufficient to meet their Pi requirements. The mutant strain appears to require more Pi in the presence of iron, leading to an increase in the expression of the alkaline phosphatase (PhoX). Additionally, iron is known to be necessary for the activity of PhoX (Monds et al., 2006). In the absence of iron, PhoX expression decreased in the mutant strain compared to the wild type. This result is consistent with those obtained with the S. lividans ppk mutant (Le Maréchal et al., 2013). In the presence of iron, the expression of the superoxide dismutase (SOD) enzyme was increased in the ∆ ppk strain, which was already under oxidative stress. Under the same conditions, the expression of the SOD was found to be decreased in the WT. These findings are consistent with those of Rodríguez-García et al. (2007). The low expression of catalase in the ∆ ppk strain in the presence of iron may be attributed to the fact that the alkyl hydroperoxide reductase (Ahp) enzyme responds to the presence of hydrogen peroxide before catalase (Seaver & Imlay, 2001). Additional experiments are required to confirm this hypothesis. The ∆ ppk strain appears to effectively utilize the antioxidant agent mycothiol to reduce the harmful effects of ROS compounds. This is due to the increased expression of myo-inositol-phosphate synthase, which is involved in mycothiol synthesis, in the mutant strain grown in the iron-supplemented medium. A significant increase was seen in some proteins related to translation in the mutant strain in the presence or absence of iron. Without iron, Δ ppk strain increased its energy metabolism compared to the wild type, so it was expected to see some increase in the translation process. When we compare the results of the mutant strain grown in the absence and presence of iron, we see that iron does not affect energy metabolism, but it does affect the translation in the mutant strain. Especially some of these proteins were strongly expressed by the mutant strain in the presence of iron. Under oxidative stress conditions, the presence of iron triggers the degradation of rRNAs, which are the building blocks of the ribosome and directly affect translation (Smethurst et al., 2020). Since Δ ppk is under oxidative stress and this stress increases with the presence of iron, it is not surprising to see an increase in the abundance of some translation related proteins. S16 is known to increase the stability of the ribosome, it was thought that the cell increases the synthesis of this and other related proteins to continue translation under stress conditions. Under oxidative stress, polyP is known to work together with chaperones to protect misfolded and damaged proteins from degradation (Gray & Jakob, 2015). The ∆ ppk strain seems to be under stress and attempts to fold denatured protein, repair damaged proteins and escape protein aggregation by increasing the expression of the chaperones and proteases in the absence of iron. In support of our findings, Varela et al. (2010) showed that Pseudomonas sp. B4 strain, which cannot produce polyP, also increased the expression of chaperones involved in protein folding. Similarly, Susin et al. (2006) reported that the activity of the GroEL-GroES chaperone system increased under oxidative stress conditions. In the presence of iron, the mutant strain reduced the expression of protein folding proteins and proteases compared to that grow without iron supplementation. In rich R2YE medium without iron supplementation, this strain decreased the expression of proteins related to energy metabolism but increased the expression of those related to translation. Although an increase in translation may have resulted in an increase in the expression of proteins related to protein folding, the opposite effect was observed. The presence of iron did not cause significant changes in the protein profile of the WT strain. This is attributed to the fact that Streptomyces is commonly found in iron-abundant environments and developed various adaptation mechanisms in this context (Choi et al., 2022). In summary, the protein profile of the ∆ ppk strain, which lacks energy and phosphate reservoir, showed significant differences compared to the wild type, independent of iron. The strain exhibited increased utilization of central energy pathways and proteins involved in protein folding, as well as some proteases. These findings highlight the importance of polyphosphate in maintaining cellular metabolism and homeostasis. Compared to the wild type strain, the mutant strain appears to increase the expression of proteins associated with phosphate metabolism to compensate for the absence of polyP in the presence of iron, in an attempt to increase the amount of intracellular free phosphate. It is possible that inorganic phosphate binds to iron, preventing its toxic effect. Recently, Pi et al. (2023) demonstrated the presence of iron-phosphate granules (ferrosomes) in a gram-positive bacterium, C. difficile . In addition, the mutant strain showed an upregulation of specific proteins related to translation and protein folding when exposed to iron, compared to the wild type strain. Compared to the iron-free medium, we see that the ∆ ppk strain reduces the expression of some enzymes involved in energy generation as a strategy to decrease oxidative stress in the presence of iron. Furthermore, the mutant strain decreases the expression of not only energy metabolism proteins but also most of the proteins identified in this study that are involved in other metabolisms. Further research is necessary to confirm whether this strain exhibits a complete stringent response and slows down its growth. Declarations Acknowledgements We would like to thank Dr. Melike Dinç (İzmir Institute of Technology) for her support in 2D gels, quantification, and identification of protein spots. Data availability All data from this study are presented in this paper. Funding This work was supported by a research grant (BAP-2018-5237) from Yıldırım Beyazıt University, Turkey. Conflict of interest The authors declared no conflict of interest. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4107881","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":286197989,"identity":"6ae6a402-ec68-48ad-be45-cc5cc0b1cf44","order_by":0,"name":"Şerif Yılmaz","email":"","orcid":"","institution":"İstanbul Arel Üniversitesi: Istanbul Arel Universitesi","correspondingAuthor":false,"prefix":"","firstName":"Şerif","middleName":"","lastName":"Yılmaz","suffix":""},{"id":286197991,"identity":"25686158-c481-45ff-acdf-d661154ff50b","order_by":1,"name":"Filiz Yeşilırmak","email":"","orcid":"","institution":"Ankara Yıldırım Beyazıt Üniversitesi: Ankara Yildirim Beyazit Universitesi","correspondingAuthor":false,"prefix":"","firstName":"Filiz","middleName":"","lastName":"Yeşilırmak","suffix":""},{"id":286197993,"identity":"05512d96-78ee-4c2c-95be-610853fae05a","order_by":2,"name":"Sedef Tunca","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAy0lEQVRIiWNgGAWjYJACiYQDNhAWDwla0iCqgYQEcVoYDhwmQYt8/+KDNx6cOW+3XyKB8cHbNoY68wYCWgxuPEu2SLhxO7lHIoHZcG4bg4TMAUJaJM6YSSR8uJ3MI5HAJs0L1ELQZfIzzn8DajkH0sL+mygtDOd72CQSbhywA9nCTJQWgxtsxhYJZ5ITeM48bJacc05CcgZBh/UffnjzxzE7e/b25IMf3pTZ8BN2mEQCmEpsYGBsYCAuJvkPgCl7IpSOglEwCkbBSAUAD70+Pc1ZMJQAAAAASUVORK5CYII=","orcid":"","institution":"Gebze Technical University: Gebze Teknik Universitesi","correspondingAuthor":true,"prefix":"","firstName":"Sedef","middleName":"","lastName":"Tunca","suffix":""}],"badges":[],"createdAt":"2024-03-15 12:44:19","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4107881/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4107881/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s11756-024-01753-y","type":"published","date":"2024-07-19T16:13:28+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":54079252,"identity":"5d0e9a12-4805-4775-915e-9495d62ebd55","added_by":"auto","created_at":"2024-04-04 09:28:27","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1419216,"visible":true,"origin":"","legend":"\u003cp\u003eRepresentative 2-D SDS gels of proteins extracted from \u003cem\u003eS. coelicolor\u003c/em\u003e (WT) grown in R2YE (a), WT\u003cem\u003e \u003c/em\u003egrown in R2YE+FeCI\u003csub\u003e3\u003c/sub\u003e (b), Δ\u003cem\u003eppk \u003c/em\u003egrown in R2YE (c), and Δ\u003cem\u003eppk\u003c/em\u003e grown in R2YE+FeCI\u003csub\u003e3 \u003c/sub\u003e(d). Representative spots indicated by arrows are numbered and correspond to the proteins listed in Table.\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-4107881/v1/eaae1a0f4f6735fddf780124.png"},{"id":54079250,"identity":"f8993935-b73f-4111-b3e7-ae62d7ff6057","added_by":"auto","created_at":"2024-04-04 09:28:26","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":983034,"visible":true,"origin":"","legend":"\u003cp\u003eInteraction networks of proteins analysed in \u003cem\u003eS. coelicolor\u003c/em\u003e. The list of protein identities obtained was entered into the STRING database (STRING version 12.0) to define known and predicted functional networks.\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-4107881/v1/4302bb5655d800ae1ee00b9e.png"},{"id":54079251,"identity":"11e0a1c6-fd07-4fe5-bada-9611280fe3ff","added_by":"auto","created_at":"2024-04-04 09:28:27","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":405256,"visible":true,"origin":"","legend":"\u003cp\u003eSummary of differentially expressed proteins of \u003cem\u003eS. coelicolor\u003c/em\u003e Δ\u003cem\u003eppk \u003c/em\u003egrown in R2YE (a) and iron-supplemented R2YE (b), \u003cem\u003ep\u003c/em\u003e-value volcano plot of increased and decreased proteins in \u003cem\u003eS. coelicolor\u003c/em\u003e Δ\u003cem\u003eppk\u003c/em\u003e and WT with and without iron (c). Quantile and min-max normalised \u003cem\u003ep\u003c/em\u003e-values are combined into volcano plots. (blue spots: increase, red spots: decrease, gray spots: no significant difference).\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-4107881/v1/c801a084b3cf860672fc1a67.png"},{"id":54079248,"identity":"5c4d111f-ce31-4a8d-b963-9a52d26ba6d7","added_by":"auto","created_at":"2024-04-04 09:28:25","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":1581646,"visible":true,"origin":"","legend":"\u003cp\u003eHeatmap of identified proteins. Comparison of protein abundance of Δ\u003cem\u003eppk\u003c/em\u003e with WT in the absence of iron (a), Δ\u003cem\u003eppk\u003c/em\u003ewith WT in the presence of iron (b), Δ\u003cem\u003eppk\u003c/em\u003e grown in the presence and absence of iron (c), WT grown in the presence and absence of iron (d). The scale represents log2 values.\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-4107881/v1/ff6dacdefb32369d9a855c5f.png"},{"id":61596724,"identity":"f6c72d06-9088-4f1b-bf0d-bebf7f8fae04","added_by":"auto","created_at":"2024-08-01 17:29:23","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":5943910,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4107881/v1/505df654-21ba-4f27-b536-14efe29985c6.pdf"},{"id":54079249,"identity":"b2e531fb-0aeb-49ab-827d-c3c442f80e51","added_by":"auto","created_at":"2024-04-04 09:28:26","extension":"xlsx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":42454,"visible":true,"origin":"","legend":"","description":"","filename":"SUPPLEMENTARY.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-4107881/v1/b99d49c6f95900f0b5083cef.xlsx"}],"financialInterests":"","formattedTitle":"A Proteomic Study to Elucidate Molecular Relationships Between Iron, Oxidative Stress and Polyphosphate in Streptomyces coelicolor A3(2)","fulltext":[{"header":"Introduction","content":"\u003cp\u003e \u003cem\u003eStreptomyces\u003c/em\u003e species are well known for their ability to synthesize valuable secondary metabolites. To develop \u003cem\u003eStreptomyces\u003c/em\u003e-based production hosts with optimized metabolic backgrounds, we need to have a holistic view of the molecular mechanisms that orchestrate secondary metabolism. The \"cause and effect\" view must therefore be replaced by the integrative study of cellular regulatory signaling networks.\u003c/p\u003e \u003cp\u003ePhosphate (Pi) availability and the oxidative stress response are two factors able to modulate \u003cem\u003eStreptomyces\u003c/em\u003e secondary metabolism(Esnault et al., 2017). Pi limitation is a trigger of secondary metabolite production and disruption of homeostasis of intracellular reactive oxygen species (ROS) causes a redirection of metabolic fluxes (Beites et al., 2011) .\u003c/p\u003e \u003cp\u003ePi metabolism in \u003cem\u003eStreptomyces\u003c/em\u003e is regulated by the two-component system PhoRP (Liu et. al., 2013). Under Pi starvation conditions, the membrane sensor kinase PhoR phosphorylates the response regulator PhoP that controls the expression of more than 40 genes that comprise the PHO regulon and are required for phosphate scavenging and assimilation. However, while maintaining Pi homeostasis, PhoP also mediates the transient shutdown of central metabolic pathways, secondary metabolism, and morphological differentiation (Allenby et al., 2012) .\u003c/p\u003e \u003cp\u003eOver the last decade, experimental data suggests a crosstalk between Pi and ROS homeostasis. In Pi limiting conditions, an indirect PhoP-dependent transcription of oxidative stress related genes (notably \u003cem\u003eahpC\u003c/em\u003e coding for the alkyl hydroperoxide reductase, an H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e detoxifying enzyme) is observed (Rodriguez-Garcia et. al., 2007) together with enhanced sensitivity to oxidative stress (Ghorbel et. al., 2006). On the other hand, the \u003cem\u003eS. natalensis\u003c/em\u003e Δ\u003cem\u003eahpC\u003c/em\u003e strain that presented high intracellular ROS levels, displayed a delayed Pi uptake and an up-regulation of the PHO regulon, namely polyphosphate kinase gene (\u003cem\u003eppk\u003c/em\u003e) (Beites et al., 2011).\u003c/p\u003e \u003cp\u003ePolyphosphate kinase enzyme (PPK) is responsible for synthesis of an important energy and phosphate storage polymer, polyphosphate (polyP), and its transcription is regulated by PhoP. Deletion of \u003cem\u003eppk\u003c/em\u003e in \u003cem\u003eS. coelicolor\u003c/em\u003e resulted in an actinorhodin overproducing phenotype (Yalim Camci et al., 2012). However, the industrial exploitation of overproducing \u003cem\u003eStreptomyces\u003c/em\u003e strains without PPK activity is hampered by their increased sensitivity to oxidative stress (Yalim Camci et al., 2012; Ghorbel et. al., 2006).\u003c/p\u003e \u003cp\u003eWorth noting that both Pi metabolism and the oxidative stress response are two physiological processes able to modulate the availability of intracellular iron. Iron is an essential element for bacterial vegetative growth and secondary metabolism (Weinberg, 1990) and it plays a key role in the bacterial oxidative stress response either as a cofactor in ROS-sensing proteins or by the generation of the highly toxic hydroxyl radical \u003cem\u003evia\u003c/em\u003e the Fenton reaction (Imlay, 2013). Although cells have ROS-degrading enzymes such as superoxide dismutases, catalases, alkylhydroperoxidases, etc. (Cornelis et al., 2011), polyP also prevents oxidative stress by inhibiting Fenton reactions via chelating iron (Gray \u0026amp; Jakob, 2015). At the same time, polyP mediates the appropriate refolding of the damaged proteins in oxidative stress conditions (Gray et al., 2014). Independent from polyP, it has recently been shown that iron is stored in gram-positive \u003cem\u003eClostridioides difficile\u003c/em\u003e in the form of \"iron-phosphate granules\" (Pi et. al., 2023).\u003c/p\u003e \u003cp\u003eIn recent years, significant progress has been made in the study of the molecular mechanisms controlling secondary metabolism in \u003cem\u003eStreptomyces\u003c/em\u003e. However, there is still insufficient information on the interactions between different regulatory mechanisms. This study aims to elucidate the metabolic relationship between oxidative stress, iron, and Pi metabolism of \u003cem\u003eS. coelicolor\u003c/em\u003e by comparative proteomic analyses of the wild-type and Δ\u003cem\u003eppk\u003c/em\u003e strains grown in iron-containing and iron-free media.\u003c/p\u003e"},{"header":"Material and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eMedia and growth conditions\u003c/h2\u003e \u003cp\u003e \u003cem\u003eS. coelicolor\u003c/em\u003e A3(2) and Δ\u003cem\u003eppk\u003c/em\u003e strains were grown in TSB and R2YE media at 30\u0026deg;C (Kieser et al., 2000). For protein isolation, seed cultures were prepared by growing the bacteria in TSB. After centrifugation at 1000\u0026times;g for 15 min. bacterial pellets were weighed and equal amounts from each strain were inoculated into R2YE broth and were grown for 48 hours at 30\u0026deg;C and 250 rpm. The presence of 0.0025 mM FeCl\u003csub\u003e3\u003c/sub\u003e in the R2YE medium was ignored and used as the iron-free medium. R2YE medium was supplemented with 100 mM FeCl\u003csub\u003e3\u003c/sub\u003e to obtain an iron-containing medium. All the flasks were washed with 8M HCI to get rid of iron contamination, rinsed, and autoclaved.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eProtein Isolation\u003c/h2\u003e \u003cp\u003eBacterial cells, grown for 48 hours at 30\u0026deg;C, 250 rpm, were collected by centrifugation at 1000\u0026times;g for 15 min. and the proteins were isolated according to Faurobert et al. (2007). Each pellet was dissolved in 2 mL extraction buffer (500 mM Tris-HCl, 50 mM EDTA, 700 mM sucrose, 100 mM KCl, pH 8.0, 1X protease inhibitor cocktail) and the cells were lysed by sonication (15 s on/30 s off, 50% of amplitude) for 5 minutes. After incubation on ice for 10 minutes, an equal volume of Tris-buffered phenol (pH8.0, Sigma) was added and the solution was shaken at room temperature for 10 min. Then the sample was centrifuged at 3200\u0026times;g for 10 min and the supernatant was carefully transferred to the new tube. After adding 3 mL extraction buffer to the sample, it was vortexed and shaken for 3 min at room temperature. Then the sample was centrifuged at 3200\u0026times;g for 10 min. and 4 volumes of precipitation buffer (0.1 M ammonium acetate dissolved in methanol) was added onto the supernatant and incubated at -20\u003csup\u003e\u0026deg;\u003c/sup\u003eC for at least 1 day. Then the proteins were precipitated by centrifugation at 3200\u0026times;g for 10 min. and they were treated 3 times with cold precipitation buffer and once with cold acetone. Isolated proteins were visualized on SDS-PAGE (Laemmli, 1970), and their concentrations were calculated by the Bradford method (Bradford, 1976) using bovine serum albumin as a control.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eProtein Separation by 2D-PAGE\u003c/h2\u003e \u003cp\u003e400 \u0026micro;g of protein sample was mixed with 3 ml rehydration buffer (7M urea, 2 M thiourea, 4% w/v CHAPS, 0.46% w/v DTT) and loaded on IPG strips (17 cm, pH 3\u0026ndash;10 nonlinear gradient, Bio-Rad, USA). IPG strips were first passively rehydrated for 2 hours and then actively rehydrated with 50 V current for 16 hours. Isoelectric focusing was carried out at 20\u0026deg;C on the IPGphor to be a total of 6000V. The strips were incubated in equilibration solution (6 M urea, 30% w/v glycerol, 2% w/v SDS, 50 mM Tris, pH 8.8) containing 1% w/v DTT, followed by 15 minutes incubation in 4% w/v iodoacetamide equilibration solution. After isoelectric focusing, the second dimension was performed in 12% polyacrylamide gels. The gels were stained with colloidal Coomassie brilliant blue solution and the gel image was transferred to a computer using a digital imaging system (VersaDoc MP 4000, Bio-Rad).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eImage analysis of 2D Gels\u003c/h2\u003e \u003cp\u003eFor spot detection \u0026ldquo;Spot colocalisation (ComDet) plug-in\u0026rdquo; and for quantification \u0026ldquo;pixel intensity\u0026rdquo; programs in ImageJ (Fiji), were used. ImageJ calculates the average pixel value in areas with grayscale values ranging from 0 (black) to 255 (white). Three biological replicates of each sample (WT and Δ\u003cem\u003eppk\u003c/em\u003e), grown in iron-free and iron supplemented media, resulted in 12 SDS PAGE gels. The result for each spot is expressed as the ratio between Δ\u003cem\u003eppk\u003c/em\u003e and WT spot intensity; and between iron-supplemented and iron-free medium for Δ\u003cem\u003eppk\u003c/em\u003e and WT.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eStatistical Analysis of Variations in Protein Abundance\u003c/h2\u003e \u003cp\u003eNormalisation is used as a technique used to adjust data from different samples in order to eliminate unwanted variability and improve comparability between datasets (Carvalho et al., 2024). In this study, we performed the quantile normalisation and minimum-maximum scaling normalisations performed by Zhao et al. (2020) on three replicates obtained in R2YE and iron supplemented R2YE for both Δ\u003cem\u003eppk\u003c/em\u003e and WT.\u003c/p\u003e \u003cp\u003eStatistical analysis according to Augillian et al. (2020) was used. After data normalisation, fold change values were calculated, and the \u003cem\u003eF\u003c/em\u003e-test was used for analysis of variation. The \u003cem\u003eF\u003c/em\u003e-test was evaluated at a significance level of \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05. If the \u003cem\u003eF\u003c/em\u003e-test is p\u0026thinsp;\u0026lt;\u0026thinsp;0.05, the variation is unequal (heteroskedastic), otherwise the variation is equal (homoskedastic). Independent two-sample \u003cem\u003et\u003c/em\u003e-test was performed according to these two variation conditions. The results of the two-sample \u003cem\u003et\u003c/em\u003e-test were evaluated at a significance level of \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05 to assess the expression differences of the proteins.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eProtein identification by matrix-assisted laser desorption/ionization tandem time of flight (MALDITOF-TOF)\u003c/h2\u003e \u003cp\u003eSpots were cut out of the gel and washed two times with a solution of 50% (v/v) methanol and 5% acetic acid until they became colorless. Then they were dehydrated with acetonitrile (ACN), treated with a solution of 10 mM DTT in 100 mM NH\u003csub\u003e4\u003c/sub\u003eHCO\u003csub\u003e3\u003c/sub\u003e for 30 minutes at room temperature, and subsequently alkylated with a solution of 100 mM iodoacetamide in 100 mM NH\u003csub\u003e4\u003c/sub\u003eHCO\u003csub\u003e3\u003c/sub\u003e for 30 minutes in dark. Following further dehydration with ACN and rehydration with NH\u003csub\u003e4\u003c/sub\u003eHCO\u003csub\u003e3\u003c/sub\u003e, the gel fragments were digested with 30 \u0026micro;L of trypsin solution (20 ng/\u0026micro;L prepared in 100 mM NH\u003csub\u003e4\u003c/sub\u003eHCO\u003csub\u003e3\u003c/sub\u003e) and incubated at 37\u0026deg;C overnight. The peptides were then extracted twice by using a solution of 5% formic acid in 50% ACN and then desalted with ZipTip. Mass spectrometry (MS) analysis was carried out by using a MALDI-TOF-TOF instrument (Bruker Autoflex III Smartbeam, USA), and the obtained spectra were processed and analyzed with the BioTools software (Bruker Daltonics, USA). Database searching was conducted separately using an in-house MASCOT server (Matrix Science, London, UK).\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eThe effect of iron on the proteome profile of \u003cem\u003eS. coelicolor\u003c/em\u003e A3(2) (WT) and \u0026Delta;\u003cem\u003eppk\u003c/em\u003e mutant strains was studied. The cells were grown in liquid R2YE and R2YE supplemented with 100 mM FeCl\u003csub\u003e3\u003c/sub\u003e. Proteins were isolated from 48-hour grown cells to make a comparison before toxic levels of ROS accumulate in the presence of iron. Isolated proteins were run on 2D gels and a total of 12 SDS PAGE gels of WT and \u0026Delta;\u003cem\u003eppk\u003c/em\u003e strains of 3 biological replicates were obtained. A representation of the protein gels for the two strains is shown in Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e\n\u003cp\u003eProtein identification was performed by protein fingerprinting, and out of 78 spots, 66 were successfully identified by using the fully sequenced and annotated genome of \u003cem\u003eS. coelicolor\u003c/em\u003e (Bentley et al., 2002), while 12 remained unidentified. Of the 66 spots identified, 37 represented distinct proteins. These proteins were classified into 6 categories (energy metabolism, phosphate metabolism, oxidative stress metabolism, translation, chaperone systems, others) and protein interaction map was generated using the STRING database (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eIt was observed that in the R2YE medium, the expression of 16 proteins was increased, 3 were decreased and 18 were not considerably altered in the \u0026Delta;\u003cem\u003eppk\u003c/em\u003e strain compared to the WT strain (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e, Table). In the iron-supplemented medium, the expression of 12 proteins increased, 6 decreased and 19 showed no significant change in the mutant strain compared to the WT strain (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e; Table).\u003c/p\u003e\n\u003cp\u003eAccording to the protein profile of the \u0026Delta;\u003cem\u003eppk\u003c/em\u003e strain grown in iron-free and iron-supplemented medium, the expression of 6 proteins was found to increase, 18 of them decreased and 13 showed no significat change in the presence of iron (Table).\u003c/p\u003e\n\u003cp\u003eIn the WT strain grown in iron-supplemented medium compared to iron-free medium, the expression of 6 proteins increased, 11 decreased and 20 proteins were not significantly changed (Table).\u003c/p\u003e\n\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e\n \u003ch2\u003eEnergy Metabolism\u003c/h2\u003e\n \u003cp\u003eCompared to the WT strain, dihydrolipoamide dehydrogenase (spot 4), 2-oxoglutarate dehydrogenase (spot 12), and dihydrolipoamide succinyltransferase (spot 27) proteins were abundant in the \u0026Delta;\u003cem\u003eppk\u003c/em\u003e strain in R2YE medium (Table, Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). These three enzymes form a highly conserved \u0026ldquo;2-oxoglutarate dehydrogenase complex\u0026rdquo; which plays an important role in cellular energy metabolism by catalysing the conversion of 2-oxoglutarate to succinyl-CoA and NADH (+\u0026thinsp;H\u003csup\u003e+\u003c/sup\u003e) in the tricarboxylic acid cycle (Chen et al., 2019). This multi-enzyme system also protects against oxidative stress through interactions with alkyl hydroperoxidases (AhpC and AhpD) (Su et al., 2019).\u003c/p\u003e\n \u003cp\u003ePhosphoenolpyruvate carboxykinase (PEPCK) (spot 13), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (spot 8) proteins were also found to be increased in \u0026Delta;\u003cem\u003eppk\u003c/em\u003e strain in R2YE medium (Table, Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). PEPCK is a crucial enzyme involved in regulating the carbon flow within the central metabolism. It captures CO\u003csub\u003e2\u003c/sub\u003e and incorporates it into PEP to generate oxaloacetate or catalyze the reverse reaction (Chiba et al., 2015). GAPDH is responsible for catalyzing the sixth step of glycolysis and also contributes to gluconeogenesis.\u003c/p\u003e\n \u003cp\u003eTransketolase (spot 15) was another enzyme whose expression was elevated in the \u0026Delta;\u003cem\u003eppk\u003c/em\u003e strain compared to WT in the absence of iron. Its function is to link glycolysis and the pentose phosphate pathway (Vimala \u0026amp; Harinarayanan, 2016) (Table, Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003eUpon the addition of iron to the medium, the mutant cell showed a decrease in energy metabolism when compared to the iron-depleted conditions. This suggests that the cell is attempting to stabilize the amount of ROS that has already formed due to Fenton reactions, as energy metabolism is one of the primary sources of ROS in cells.\u003c/p\u003e\n \u003cp\u003eIn the iron-supplemented medium, only glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein was found to be increased in \u0026Delta;\u003cem\u003eppk\u003c/em\u003e compared to the WT strain. Similarly, the presence of iron did not significantly affect the protein profile of the WT strain. However, dihydrolipoamide succinyltransferase (spot 27) was found to be increased, while fructose bisphosphate aldolase (spot 32) was found to be decreased.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n \u003ch2\u003ePhosphate Metabolism\u003c/h2\u003e\n \u003cp\u003eCompared to WT, PhoP (spot 17), PhoX (spot 23) and PhoU (spot 26) proteins were found to be abundant in the \u0026Delta;\u003cem\u003eppk\u003c/em\u003e strain in iron supplemented medium. In the absence of iron, only PhoX protein was found to be decreased (Table, Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). PhoP controls more than 40 genes involved in phosphate scavenging and assimilation. PhoU is the modulator of phosphate-specific transport system and PhoX is an alkaline phosphatase that functions under Pi-limiting conditions (Mart\u0026iacute;n \u0026amp; Liras, 2021). The presence of iron caused a slight decrease in the expression of PhoP and PhoX in WT.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n \u003ch2\u003eOxidative stress\u003c/h2\u003e\n \u003cp\u003eMyo-inositol-phosphate synthase, superoxide dismutase (SOD) and catalase play essential roles in oxidative stress resistance. SOD and catalase enzymes catalyze the conversion of reactive oxygen species into harmless forms (Kanth et al., 2011). Myo-inositol-phosphate synthase the conversion of D-glucose 6-phosphate to 1L-myo-inositol 1-phosphate (Geiger \u0026amp; Jin, 2006), which is the precursor of an antioxidant agent mycothiol (Chen et al., 2019). Compared to WT, SOD (spot 20) and myo-inositol phosphate synthase (spot 7) were found to be more abundant and catalase (spot 31) was less abundant in the \u0026Delta;\u003cem\u003eppk\u003c/em\u003e strain in the presence of iron. Presence of iron caused a decrease in the expression of both SOD and myo-inositol phosphate synthase in the mutant strain, compared to that grew in the absence of iron. In the WT strain, SOD was found to be less abundant and catalase was found to be more abundant in the presence of iron compared to the WT strain in the R2YE (Table, Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\n \u003ch2\u003eTranslation\u003c/h2\u003e\n \u003cp\u003eCompared to the WT strain, ribosomal proteins S1 (spot 2) and S10 (spot 39) were found to be abundant, while ribosomal protein L17 (spot 24) and Elongation factor Tu-1 (spot 37) were found to be decreased in \u0026Delta;\u003cem\u003eppk\u003c/em\u003e strain in R2YE medium.\u003c/p\u003e\n \u003cp\u003eIn the iron-supplemented medium, ribosomal proteins S10 (spot 39), S16 (spot 34), L10 (spot 35), L17 ribosomal proteins (spot 24) were found to be up-regulated together with proline tRNA ligase (spot 25) in the \u0026Delta;\u003cem\u003eppk\u003c/em\u003e strain, compared to WT. The increase in the expression level of ribosomal protein S16 was very pronounced (log2 fold change 4,3). The presence of iron caused an increase in the expression of ribosomal protein S1; and a decrease in L17 and S10 in WT (Table, Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). \u0026Delta;\u003cem\u003eppk\u003c/em\u003e grown in iron, expressed more L17, L10, S16, Elongation factor Tu-1 and proline-tRNA ligase than those grown in iron-free medium. Proline-tRNA ligase catalyses the addition of proline to tRNA and elongation factor Tu-1 brings the aminoacyl tRNAs to the aminoacyl region of the ribosome (Rodnina, 2016).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n \u003ch2\u003eProtein Folding and Proteases\u003c/h2\u003e\n \u003cp\u003eAmong the major chaperone proteins, the trigger factor (TF) (spot 1), GroES (spot 11) and GroEL (spot 33) proteins, the Clp family protease (spot 14) and tricorn protease (spot 29) proteins were abundant in \u0026Delta;\u003cem\u003eppk\u003c/em\u003e compared to WT in R2YE medium. It is known that differential expression of Clp protease genes affects the differentiation process in \u003cem\u003eStreptomyces\u003c/em\u003e (Bellier \u0026amp; Mazodier, 2004). On the other hand, the tricorn protease is involved in the recycling of proteins within the cell (Tamura et al., 2001).\u003c/p\u003e\n \u003cp\u003eIn iron supplemented medium, GroES was found to be increased and Clp family protease and tricorn protease were decreased in \u0026Delta;\u003cem\u003eppk\u003c/em\u003e strain compared to WT. Similarly, except for GroES, TF, GroEL, the Clp protease and tricorn protease were less abundant in the \u0026Delta;\u003cem\u003eppk\u003c/em\u003e strain in the presence of iron compared to iron-free conditions. Clp family protease and tricorn protease were increased, GroEL and GroES on the other hand was decreased in WT in the iron supplemented medium (Table, Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\n \u003ch2\u003eOther Proteins\u003c/h2\u003e\n \u003cp\u003eN-acetylneuraminate synthase (spot 9) and N-acylneuraminate cytidylyltransferase (spot 3) proteins were abundant in \u0026Delta;\u003cem\u003eppk\u003c/em\u003e strain in R2YE medium compared to WT. Both enzymes participate in aminosugar metabolism, specifically related with cell wall synthesis (Bravo et al., 2001). Ribbon-helix-helix protein (spot 19), which is a transcription factor that plays an important role in the regulation of metal uptake, amino acid biosynthesis, cell division, etc. (Schreiter \u0026amp; Drennan, 2007), was found to be also slightly increased in \u0026Delta;\u003cem\u003eppk\u003c/em\u003e compared to WT in R2YE medium. In the presence of iron, the expression of this protein, as well as N-acetylneuraminate synthase, N-acylneuraminate cytidylyltransferase, and glutamate dehydrogenase (spot 28), were found to be decreased in the \u0026Delta;\u003cem\u003eppk\u003c/em\u003e strain compared to iron-free conditions. Glutamate dehydrogenase plays a major role in amino acid metabolism by catalysing the reversible process of oxidative deamination, converting glutamate to \u0026alpha;-ketoglutarate (Smith et al., 2019).\u003c/p\u003e\n \u003cp\u003eCompared to the WT strain, glutamate dehydrogenase and RNA polymerase subunit beta\u0026apos; (spot 30) were found to be reduced in \u0026Delta;\u003cem\u003eppk\u003c/em\u003e in iron supplemented R2YE. RNA polymerase subunit beta\u0026apos; is one of the major subunits of the RNA polymerase holoenzyme (Sutherland \u0026amp; Murakami, 2018).\u003c/p\u003e\n \u003cp\u003eWhen all the results were analysed together, it was observed that compared to the wild type strain, the levels of proteins involved in energy metabolism and the chaperone system were increased in the mutant strain in iron-free medium, whereas the levels of SOD and mycothiol, which are elements of the antioxidant system, were increased in iron-containing medium, together with proteins involved in phosphate metabolism and translation.\u003c/p\u003e\n \u003cp\u003eWhen comparing the protein profiles of the mutant strain in the presence and absence of iron, it is evident that the strain has difficulty coping with the iron in the absence of polyphosphate. In the presence of iron, it only increases the expression of translational proteins and PhoX, while decreasing the expression of most of the proteins, identified in this study that are involved in other metabolisms.\u003c/p\u003e\n \u003cp\u003eWhile the presence of iron did not significantly alter the total proteome of the wild-type strain, it did cause a decrease in the expression of PhoP, PhoX, Fructose-bisphosphate aldolase, SOD, ribosomal proteins L17 and S10. The concentration of iron used in this study did not cause toxicity in the wild-type strain. On the contrary, it caused a decrease in the expression of UspA, a putative stress protein.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eTable.\u003c/strong\u003e Differentially expressed proteins (\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e: increase,\u0026nbsp;\u0026darr;: decrease,\u0026nbsp;─: no significant differences)\u003c/p\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"996\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003eSpot No\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003e\u003cstrong\u003eProtein Name\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026Delta;ppk/\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eWT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003eFold Change\u003cbr\u003e\u0026nbsp;log2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026Delta;ppkFe/\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eWTFe\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003eFold Change\u003cbr\u003e\u0026nbsp;log2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026Delta;ppkFe/\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026Delta;ppk\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003eFold Change\u003cbr\u003e\u0026nbsp;log2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003eWTFe/\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eWT\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003eFold Change\u003cbr\u003e\u0026nbsp;log2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003e\u003cstrong\u003eGene\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003e\u003cstrong\u003eUniProt\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003eMw\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e\u003cstrong\u003epI\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e\u003cstrong\u003ePMF\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003eMSMS\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"100%\" colspan=\"17\"\u003e\n \u003cp\u003e\u003cstrong\u003eEnergy Metabolism\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e4\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eDihydrolipoamide dehydrogenase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,8*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,4*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO2180\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9S2Q6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e51536\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e8\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eGlyceraldehyde-3-phosphate dehydrogenase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,6*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,4*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u0026nbsp;─\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO7511\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ93J08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e35003\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e12\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003ePutative 2-oxoglutarate dehydrogenase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,2*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,6*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO5281\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9FBR4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e139292\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e13\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003ePhosphoenolpyruvate carboxylase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,6*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,1*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO3127\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9RNU9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e101349\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e15\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eTransketolase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,5*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u0026nbsp;─\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO1935\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9XAC1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e75085\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e27\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eDihydrolipoamide succinyltransferase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,7*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,6*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,5*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO2181\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9S2Q5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e59000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e4,57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e32\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eFructose-bisphosphate aldolase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,8*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,7*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO3649\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9X8R6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e37018\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"100%\" colspan=\"17\"\u003e\n \u003cp\u003e\u003cstrong\u003ePhosphate Metabolism\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e17\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003ePutative response regulator (PhoP)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,6*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u0026nbsp;─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,6*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO4230\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9L0R1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e24770\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e23\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003ePhoX\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u0026darr;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,0*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,5*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,4*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,1*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO3790\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9F2J1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e75179\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e6,21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e26\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003ePhosphate-specific transport system accessory protein PhoU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e1,0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,6*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,2*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO4228\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ8CJU3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e25441\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e4,93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"100%\" colspan=\"17\"\u003e\n \u003cp\u003e\u003cstrong\u003eOxidative stress\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e7\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eMyo-inositol-1-phosphate synthase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,6*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,6*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-1,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO3899\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9X8T5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e39433\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e20\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eSuperoxide dismutase [Fe-Zn]\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,5*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,8*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,2*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO2633\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eO51917\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e23513\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e31\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eCatalase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,3*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,1*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO0379\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9RJK9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e55139\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"100%\" colspan=\"17\"\u003e\n \u003cp\u003e\u003cstrong\u003eTranslation\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003e30S ribosomal protein S1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,5*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,3*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,6*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,3*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO1998\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9S2K5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e55001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e4,54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e6\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003e30S ribosomal protein S2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO5624\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eO31212\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e33602\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e24\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003e50S ribosomal protein L17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,3*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,4*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,4*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,4*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO4730\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eO86775\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e18120\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e9,42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e25\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eProline--tRNA ligase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,6*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,9*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO5699\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9KYR6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e61443\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e4,98\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e34\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003e30S ribosomal protein S16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e4,3*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,1*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-2,7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO5591\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eO69879\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e15187\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e9,25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e35\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003e50S ribosomal protein L10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,0*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,5*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO4652\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eP41103\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e18635\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e8,78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e37\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eElongation factor Tu-1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-2,0*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,9*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSC04662\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eP40174\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e43811\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e39\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003e30S ribosomal protein S10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,0*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,2*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,4*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO4701\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eP66337\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e11571\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e9,3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"100%\" colspan=\"17\"\u003e\n \u003cp\u003e\u003cstrong\u003eProtein folding and Proteases\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eTrigger factor\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,5*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,4*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e1,0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO2620\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9F314\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e51137\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e4,31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e11\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eCo-chaperonin GroES\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,8*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,5*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,1*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO4761\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eP0A345\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e10940\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e4,65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e14\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003ePutative Clp-family ATP-binding protease\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,7*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,0*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,4*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,3*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO3373\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9S6T8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e92957\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e29\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eTricorn protease homolog 1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,1*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-2,1*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-2,4*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,9*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO2549\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9RDE2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e115577\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e33\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eChaperonin GroEL 2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,6*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,6*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,1*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO4296\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9KXU5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e56795\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e4,79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"100%\" colspan=\"17\"\u003e\n \u003cp\u003e\u003cstrong\u003eOthers\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eN-acylneuraminate cytidylyltransferase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e2,0*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,7*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO4880\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9AK46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e44514\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003ePhage Tail Protein\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO4252\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9L0N9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e16527\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e4,86\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e9\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eN-acetylneuraminate synthase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,4*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,8*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO4880\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9AK45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e34716\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e10\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eUspA domain-containing protein\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO0200\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9RI46\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e32237\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e16\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003ePutative 3-oxoacyl-[acyl-carrier protein] reductase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,6*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO6282\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ93S07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e27414\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e18\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eConserved hypothetical membrane protein\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,6*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,8*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO3967\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ93J39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e25827\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e19\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eRibbon-helix-helix protein, CopG family\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026uarr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,3*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,1*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,6*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO5908\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eO54104\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e7146\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e28\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eGlutamate dehydrogenase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-2,7*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-2,7*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO2999\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ8CJY0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e183751\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e30\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eDNA-directed RNA polymerase subunit beta\u0026apos;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-3,0*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-2,3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO4655\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ8CJT1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e145199\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e6,44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e36\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003ePutative ABC transporter ATP-binding protein\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO3224\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9S6T6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e34069\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e6,22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"4.012036108324975%\"\u003e\n \u003cp\u003e\u003cstrong\u003e40\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"25.27582748244734%\"\u003e\n \u003cp\u003eUspA-Putative Stress Protein\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.413239719157472%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e0,6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.817452357071214%\"\u003e\n \u003cp\u003e─\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e-0,6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"5.4162487462387165%\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026darr;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,3*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.218655967903711%\"\u003e\n \u003cp\u003eSCO0167\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"6.118355065195587%\"\u003e\n \u003cp\u003eQ9RIZ8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e31222\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.309929789368104%\"\u003e\n \u003cp\u003e5,66\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"3.8114343029087263%\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"4.914744232698094%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"0.30090270812437314%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003csup\u003e\u0026times;\u003c/sup\u003e Fold change data showing that the comparison of protein data is significant (\u003cem\u003ep\u003c/em\u003e \u0026lt;0.05)\u003c/p\u003e\u0026nbsp;\n\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003ePolyphosphate (polyP) is a conserved polymer that has important functions in every organism. One important function of polyP is to help cells resist various stresses. \u003cem\u003eStreptomyces\u003c/em\u003e strains that are unable to produce polyP are known to have increased sensitivity to oxidative stress (Ghorbel et al., 2006; Yalim Camci et al., 2012; Le Mar\u0026eacute;chal et al., 2013). Iron is an important element in the cell, but it plays a role in the generation of oxidative stress by causing the formation of ROS compounds through Fenton reactions. In this study, comparative proteomic analyses of \u003cem\u003eS. coelicolor ppk\u003c/em\u003e mutant (∆\u003cem\u003eppk\u003c/em\u003e) and wild-type (WT) strains were performed in the presence and absence of iron to understand the relationship between polyphosphate, oxidative stress, and iron metabolisms.\u003c/p\u003e \u003cp\u003eIt was found that in the absence of poly P, which is an important energy storage polymer, the ∆\u003cem\u003eppk\u003c/em\u003e strain suffered from energy deprivation even in R2YE, a nutrient-rich medium that promotes \u003cem\u003eStreptomyces\u003c/em\u003e growth. To overcome this energy deficit, the relevant pathways were activated in the mutant strain. Presence of iron did not caused significant changes in the abundance of energy metabolism proteins in Δ\u003cem\u003eppk\u003c/em\u003e compared to WT. However ∆\u003cem\u003eppk\u003c/em\u003e strain reduced the expression of enzymes involved in energy metabolism in the presence of iron compared to the conditions without iron. \u003cem\u003eStreptomyces\u003c/em\u003e are characterized by a high utilization of the Pentose Phosphate pathway and complex-I (NADH:ubiquinone oxidoreductase) respiratory enzymes, which under normal conditions produce more ROS (Lejeune et al., 2022). The TCA cycle is also an important source of ROS metabolites (Kohanski et al., 2007). It can be hypothesized that the mutant strain, which is already under oxidative stress, tries to reduce the amount of ROS by decreasing the expression of enzymes involved in energy metabolism. This result is not surprising since \u003cem\u003eStreptomyces\u003c/em\u003e are known to change their metabolic networks under certain conditions. For example, Choi et al. (2022) showed that in \u003cem\u003eS. coelicolor\u003c/em\u003e under oxygen-limited conditions, the presence of iron altered the cell metabolism and increased the NAD/NADH\u003csup\u003e+\u003c/sup\u003e ratio supporting anaerobic growth.\u003c/p\u003e \u003cp\u003eThe expression of proteins associated with phosphate metabolism such as PhoP, PhoU, and PhoX was found to be increased in the presence of iron in the ∆\u003cem\u003eppk\u003c/em\u003e strain. It was clear that the mutant strain tried to increase the amount of free phosphate when there was iron in the environment. Phosphate metabolism in \u003cem\u003eStreptomyces\u003c/em\u003e is regulated by a two-component system called PhoR/PhoP. In this system, PhoR functions as a sensor kinase while PhoP acts as a response regulator. Other than phosphate utilization and storage, PhoP protein has also been associated with oxidative stress resistance. In a study with \u003cem\u003eS. coelicolor\u003c/em\u003e PhoP mutant, the expression of some genes related to resistance to oxidative stress was found to be reduced (Rodr\u0026iacute;guez-Garc\u0026iacute;a et al., 2007). PhoP is also involved in the positive regulation of the \u003cem\u003eppk\u003c/em\u003e gene and indirectly of polyP, which chelates iron and prevents the formation of ROS by the Fenton reaction (Beaufay et al., 2020). PhoU is another protein that has been associated with oxidative stress. It was observed that H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e sensitivity was increased in the mutant strain that could not produce PhoU protein (Ghorbel et al., 2006).\u003c/p\u003e \u003cp\u003eBacteria induce alkaline phosphatases when inorganic phosphate (Pi) is insufficient to meet their Pi requirements. The mutant strain appears to require more Pi in the presence of iron, leading to an increase in the expression of the alkaline phosphatase (PhoX). Additionally, iron is known to be necessary for the activity of PhoX (Monds et al., 2006). In the absence of iron, PhoX expression decreased in the mutant strain compared to the wild type. This result is consistent with those obtained with the \u003cem\u003eS. lividans ppk\u003c/em\u003e mutant (Le Mar\u0026eacute;chal et al., 2013).\u003c/p\u003e \u003cp\u003eIn the presence of iron, the expression of the superoxide dismutase (SOD) enzyme was increased in the ∆\u003cem\u003eppk\u003c/em\u003e strain, which was already under oxidative stress. Under the same conditions, the expression of the SOD was found to be decreased in the WT. These findings are consistent with those of Rodr\u0026iacute;guez-Garc\u0026iacute;a et al. (2007). The low expression of catalase in the ∆\u003cem\u003eppk\u003c/em\u003e strain in the presence of iron may be attributed to the fact that the alkyl hydroperoxide reductase (Ahp) enzyme responds to the presence of hydrogen peroxide before catalase (Seaver \u0026amp; Imlay, 2001). Additional experiments are required to confirm this hypothesis.\u003c/p\u003e \u003cp\u003eThe ∆\u003cem\u003eppk\u003c/em\u003e strain appears to effectively utilize the antioxidant agent mycothiol to reduce the harmful effects of ROS compounds. This is due to the increased expression of myo-inositol-phosphate synthase, which is involved in mycothiol synthesis, in the mutant strain grown in the iron-supplemented medium.\u003c/p\u003e \u003cp\u003eA significant increase was seen in some proteins related to translation in the mutant strain in the presence or absence of iron. Without iron, Δ\u003cem\u003eppk\u003c/em\u003e strain increased its energy metabolism compared to the wild type, so it was expected to see some increase in the translation process. When we compare the results of the mutant strain grown in the absence and presence of iron, we see that iron does not affect energy metabolism, but it does affect the translation in the mutant strain. Especially some of these proteins were strongly expressed by the mutant strain in the presence of iron. Under oxidative stress conditions, the presence of iron triggers the degradation of rRNAs, which are the building blocks of the ribosome and directly affect translation (Smethurst et al., 2020). Since Δ\u003cem\u003eppk\u003c/em\u003e is under oxidative stress and this stress increases with the presence of iron, it is not surprising to see an increase in the abundance of some translation related proteins. S16 is known to increase the stability of the ribosome, it was thought that the cell increases the synthesis of this and other related proteins to continue translation under stress conditions.\u003c/p\u003e \u003cp\u003eUnder oxidative stress, polyP is known to work together with chaperones to protect misfolded and damaged proteins from degradation (Gray \u0026amp; Jakob, 2015). The ∆\u003cem\u003eppk\u003c/em\u003e strain seems to be under stress and attempts to fold denatured protein, repair damaged proteins and escape protein aggregation by increasing the expression of the chaperones and proteases in the absence of iron. In support of our findings, Varela et al. (2010) showed that \u003cem\u003ePseudomonas sp.\u003c/em\u003e B4 strain, which cannot produce polyP, also increased the expression of chaperones involved in protein folding. Similarly, Susin et al. (2006) reported that the activity of the GroEL-GroES chaperone system increased under oxidative stress conditions. In the presence of iron, the mutant strain reduced the expression of protein folding proteins and proteases compared to that grow without iron supplementation. In rich R2YE medium without iron supplementation, this strain decreased the expression of proteins related to energy metabolism but increased the expression of those related to translation. Although an increase in translation may have resulted in an increase in the expression of proteins related to protein folding, the opposite effect was observed.\u003c/p\u003e \u003cp\u003eThe presence of iron did not cause significant changes in the protein profile of the WT strain. This is attributed to the fact that \u003cem\u003eStreptomyces\u003c/em\u003e is commonly found in iron-abundant environments and developed various adaptation mechanisms in this context (Choi et al., 2022).\u003c/p\u003e \u003cp\u003eIn summary, the protein profile of the ∆\u003cem\u003eppk\u003c/em\u003e strain, which lacks energy and phosphate reservoir, showed significant differences compared to the wild type, independent of iron. The strain exhibited increased utilization of central energy pathways and proteins involved in protein folding, as well as some proteases. These findings highlight the importance of polyphosphate in maintaining cellular metabolism and homeostasis.\u003c/p\u003e \u003cp\u003eCompared to the wild type strain, the mutant strain appears to increase the expression of proteins associated with phosphate metabolism to compensate for the absence of polyP in the presence of iron, in an attempt to increase the amount of intracellular free phosphate. It is possible that inorganic phosphate binds to iron, preventing its toxic effect. Recently, Pi et al. (2023) demonstrated the presence of iron-phosphate granules (ferrosomes) in a gram-positive bacterium, \u003cem\u003eC. difficile\u003c/em\u003e. In addition, the mutant strain showed an upregulation of specific proteins related to translation and protein folding when exposed to iron, compared to the wild type strain.\u003c/p\u003e \u003cp\u003eCompared to the iron-free medium, we see that the ∆\u003cem\u003eppk\u003c/em\u003e strain reduces the expression of some enzymes involved in energy generation as a strategy to decrease oxidative stress in the presence of iron. Furthermore, the mutant strain decreases the expression of not only energy metabolism proteins but also most of the proteins identified in this study that are involved in other metabolisms. Further research is necessary to confirm whether this strain exhibits a complete stringent response and slows down its growth.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u0026nbsp;\u003c/strong\u003eWe would like to thank Dr. Melike Din\u0026ccedil; (İzmir Institute of Technology) for her support in 2D gels, quantification, and identification of protein spots.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e All data from this study are presented in this paper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e This work was supported by a research grant (BAP-2018-5237) from Yıldırım Beyazıt University, Turkey.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e The authors declared no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u0026nbsp;\u003c/strong\u003eŞerif Yılmaz: Data curation, Writing- Original draft preparation, Visualization, Investigation. Filiz Yeşilırmak: Resources, Methodology Sedef Tunca: Resources, Supervision, Conceptualization, Methodology, Writing-Reviewing and Editing.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAguilan, J. T., Kulej, K., \u0026amp; Sidoli, S. (2020). Guide for protein fold change and p-value calculation for non-experts in proteomics. Molecular Omics, 16(6), 573\u0026ndash;582. https://doi.org/10.1039/D0MO00087F\u003c/li\u003e\n\u003cli\u003eAllenby N.E.E., Laing E., Bucca G., Kierzek A.M., Smith C.P. 2012. Diverse control of metabolism and other cellular processes in \u003cem\u003eStreptomyces coelicolor\u003c/em\u003e by the PhoP transcription factor: genome-wide identification of in vivo targets. Nucleic Acids Research, 4019, 9543\u0026ndash;9556. https://doi.org/10.1093/NAR/GKS766\u003c/li\u003e\n\u003cli\u003eBeaufay F., Quarles E., Franz A., Katamanin O., Wholey W.Y., Jakob U. 2020. 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Deletion of the polyP synthesising enzyme, polyP kinase (PPK), resulted in an antibiotic overproducing phenotype in \u003cem\u003eStreptomyces\u003c/em\u003e. However, the industrial use of overproducing \u003cem\u003eStreptomyces\u003c/em\u003e strains without PPK activity (∆\u003cem\u003eppk\u003c/em\u003e) is hampered by their increased sensitivity to oxidative stress. Iron plays a key role in the bacterial response to oxidative stress, and it is also an essential element for various processes in the cell. Conversely, polyP can sequester iron, reducing its bioavailability. This study aimed to elucidate the metabolic relationship between oxidative stress, iron, and polyP metabolisms in \u003cem\u003eStreptomyces coelicolor\u003c/em\u003e as an example of the communication of cellular regulatory signalling networks.\u003c/p\u003e \u003cp\u003eComparative proteomic analyses were performed on three biological replicates of wild-type and ∆\u003cem\u003eppk\u003c/em\u003e strains grown in iron-containing and iron-free media. Independent of iron, the results show that the absence of polyP significantly alters the total proteome, revealing the importance of this polymer in maintaining cellular metabolism. The mutant strain was found to have difficulties coping with the iron even in the nutrient-rich medium. Compared to the wild type in the iron-free medium, a general abundance of proteins related to energy metabolism, and protein folding was observed in ∆\u003cem\u003eppk\u003c/em\u003e. In the presence of iron, the expression of the proteins involved in translation, phosphate metabolism and the antioxidant system was increased in the mutant strain compared to the wild type. To our knowledge, this is the first study to clarify the relationship between iron, oxidative stress, and polyphosphate.\u003c/p\u003e","manuscriptTitle":"A Proteomic Study to Elucidate Molecular Relationships Between Iron, Oxidative Stress and Polyphosphate in Streptomyces coelicolor A3(2)","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-04-04 09:28:19","doi":"10.21203/rs.3.rs-4107881/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revisions","date":"2024-04-30T08:31:00+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2024-04-02T07:01:25+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-04-01T13:44:17+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-03-20T09:10:56+00:00","index":"","fulltext":""},{"type":"submitted","content":"Biologia","date":"2024-03-18T09:59:04+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"biologia","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"biol","sideBox":"Learn more about [Biologia](http://link.springer.com/journal/11756)","snPcode":"11756","submissionUrl":"https://www.editorialmanager.com/biol/default2.aspx","title":"Biologia","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"ac218913-f9ef-4504-9ef4-4e43e6661187","owner":[],"postedDate":"April 4th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-08-01T17:14:11+00:00","versionOfRecord":{"articleIdentity":"rs-4107881","link":"https://doi.org/10.1007/s11756-024-01753-y","journal":{"identity":"biologia","isVorOnly":false,"title":"Biologia"},"publishedOn":"2024-07-19 16:13:28","publishedOnDateReadable":"July 19th, 2024"},"versionCreatedAt":"2024-04-04 09:28:19","video":"","vorDoi":"10.1007/s11756-024-01753-y","vorDoiUrl":"https://doi.org/10.1007/s11756-024-01753-y","workflowStages":[]},"version":"v1","identity":"rs-4107881","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4107881","identity":"rs-4107881","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}