Formation of volutin granules in<i>Corynebacterium glutamicum</i>

Pallerla, Srinivas Reddy; Knebel, Sandra; Polen, Tino; Klauth, Peter; Hollender, Juliane; Wendisch, Volker F.; Schoberth, Siegfried M.

doi:10.1016/j.femsle.2004.11.047

Cited by 56 publications

(39 citation statements)

References 27 publications

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“…This situation is not unprecedented and, as an example, the industrially relevant Corynebacterium glutamicum was considered a poly-P Ϫ microorganism in contrast to the pathogenic Corynebacterium diphtheriae. However, poly-P inclusions were finally reported for the former species (45).…”

Section: Discussionmentioning

confidence: 99%

Accumulation of Polyphosphate in Lactobacillus spp. and Its Involvement in Stress Resistance

Alcántara

Blasco

Zúñiga

et al. 2014

Appl Environ Microbiol

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Polyphosphate (poly-P) is a polymer of phosphate residues synthesized and in some cases accumulated by microorganisms, where it plays crucial physiological roles such as the participation in the response to nutritional stringencies and environmental stresses. Poly-P metabolism has received little attention in Lactobacillus, a genus of lactic acid bacteria of relevance for food production and health of humans and animals. We show that among 34 strains of Lactobacillus, 18 of them accumulated intracellular poly-P granules, as revealed by specific staining and electron microscopy. Poly-P accumulation was generally dependent on the presence of elevated phosphate concentrations in the culture medium, and it correlated with the presence of polyphosphate kinase (ppk) genes in the genomes. The ppk gene from Lactobacillus displayed a genetic arrangement in which it was flanked by two genes encoding exopolyphosphatases of the Ppx-GppA family. The ppk functionality was corroborated by its disruption (LCABL_27820 gene) in Lactobacillus casei BL23 strain. The constructed ppk mutant showed a lack of intracellular poly-P granules and a drastic reduction in poly-P synthesis. Resistance to several stresses was tested in the ppk-disrupted strain, showing that it presented a diminished growth under high-salt or low-pH conditions and an increased sensitivity to oxidative stress. These results show that poly-P accumulation is a characteristic of some strains of lactobacilli and may thus play important roles in the physiology of these microorganisms.

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Section: Discussionmentioning

confidence: 99%

Accumulation of Polyphosphate in Lactobacillus spp. and Its Involvement in Stress Resistance

Alcántara

Blasco

Zúñiga

et al. 2014

Appl Environ Microbiol

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“…Chemical shifts were referenced to 85% orthophosphoric acid (0 ppm). Standards of P i and polyphosphate ("P68" with polymerization from 10 to 40; BK Giulini Chemie, Ladenburg, Germany) were prepared with final concentrations of 10 mM, in terms of P i (29). Signals were integrated with the MestRe Nova software (Mestrelab Research, Santiago, Spain) to quantify total intracellular P i and phosphorus-containing metabolites, e.g., phosphomonoesters such as sugar phosphates, NDP-glucose, or polyphosphate.…”

Section: Gc-(ei/ci)-tof Msmentioning

confidence: 99%

“…An overview of the biology, genetics, physiology, and application of C. glutamicum can be found in two recent monographs (3, 6). Phosphorus constitutes 1.5% to 2.1% of the cell dry weight of C. glutamicum (24), part of which can be present as polyphosphate (22,29). Several of the enzymes involved in polyphosphate metabolism have been characterized recently, such as a class II polyphosphate kinase (28), the exopolyphosphatases Ppx1 and Ppx2 (26), a polyphosphate/ATP-dependent glucokinase (25), and a polyphosphate/ ATP-dependent NAD ϩ kinase (27).…”

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confidence: 99%

Link between Phosphate Starvation and Glycogen Metabolism in Corynebacterium glutamicum , Revealed by Metabolomics

Woo

Noack

Seibold

et al. 2010

Appl Environ Microbiol

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In this study, we analyzed the influence of phosphate (P i ) limitation on the metabolism of Corynebacterium glutamicum. Metabolite analysis by gas chromatography-time-of-flight (GC-TOF) mass spectrometry of cells cultivated in glucose minimal medium revealed a greatly increased maltose level under P i limitation. As maltose formation could be linked to glycogen metabolism, the cellular glycogen content was determined. Unlike in cells grown under P i excess, the glycogen level in P i -limited cells remained high in the stationary phase. Surprisingly, even acetate-grown cells, which do not form glycogen under P i excess, did so under P i limitation and also retained it in stationary phase. Expression of pgm and glgC, encoding the first two enzymes of glycogen synthesis, phosphoglucomutase and ADP-glucose pyrophosphorylase, was found to be increased 6-and 3-fold under P i limitation, respectively. Increased glycogen synthesis together with a decreased glycogen degradation might be responsible for the altered glycogen metabolism. Independent from these experimental results, flux balance analysis suggested that an increased carbon flux to glycogen is a solution for C. glutamicum to adapt carbon metabolism to limited P i concentrations.Phosphorus is an essential nutrient for all cells and is required for, e.g., the biosynthesis of nucleotides, NAD(P)H, DNA, and RNA but also for the regulation of protein activity by phosphorylation of histidine, aspartate, serine, threonine, or tyrosine residues. A common phosphorus source is inorganic phosphate (P i ), and cells have developed mechanisms for the acquisition, assimilation, and storage of P i . When P i becomes limiting, many bacteria induce the synthesis of proteins that enable them to capture the residual P i resources more efficiently and to make alternative phosphorus sources accessible. The corresponding genes are collectively named P i starvationinducible genes, or psi genes. The P i starvation response, and in particular its regulation, has been most carefully studied in Escherichia coli (45) and Bacillus subtilis (14).We recently started to characterize the P i starvation response in Corynebacterium glutamicum, a Gram-positive soil bacterium used industrially for the production of more than two millions tons of amino acids per year, mainly L-glutamate and L-lysine (12). An overview of the biology, genetics, physiology, and application of C. glutamicum can be found in two recent monographs (3, 6). Phosphorus constitutes 1.5% to 2.1% of the cell dry weight of C. glutamicum (24), part of which can be present as polyphosphate (22,29). Several of the enzymes involved in polyphosphate metabolism have been characterized recently, such as a class II polyphosphate kinase (28), the exopolyphosphatases Ppx1 and Ppx2 (26), a polyphosphate/ATP-dependent glucokinase (25), and a polyphosphate/ ATP-dependent NAD ϩ kinase (27). The P i starvation stimulon of C. glutamicum was determined using whole-genome DNA microarrays (15). Comparison of the mRNA profiles before and at diffe...

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“…An overview of the biology, genetics, physiology, and application of C. glutamicum can be found in a recent monograph (5). Phosphorus constitutes 1.5% to 2.1% of the cell dry weight of C. glutamicum (18), part of which is present as polyphosphate (15,17,21). The phosphate starvation stimulon of C. glutamicum was determined using wholegenome DNA microarrays (13).…”

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confidence: 99%

Target Genes and DNA-Binding Sites of the Response Regulator PhoR fromCorynebacterium glutamicum

Schaaf

Bott

2007

J Bacteriol

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The two-component signal transduction system PhoRS of Corynebacterium glutamicum is involved in the phosphate (P i ) starvation response. To analyze the binding of unphosphorylated and phosphorylated PhoR to the promoters of phosphate starvation-inducible (psi) genes, this response regulator and the kinase domain of its cognate sensor, PhoS (MBP-PhoS⌬1-246), were overproduced and purified. MBP-PhoS⌬1-246 showed constitutive autophosphorylation activity, and a rapid phosphoryl group transfer from phosphorylated MBPPhoS⌬1-246 to PhoR was observed. Gel mobility shift assays revealed that phosphorylation increases the DNA-binding affinity of PhoR. The affinity of PhoRϳP to different promoters varied and decreased in the order pstSCAB > phoRS > phoC > ushA > porB > ugpA > pitA > nucH and phoH1 > glpQ1. The binding sites in front of pstSCAB and phoRS were localized at positions ؊194 to ؊176 and ؊61 to ؊43 upstream of the transcriptional start sites, respectively. Alignment of these two 19-bp binding sites revealed a high identity in the 5-terminal part, but not in the 3-terminal part. As many OmpR-type response regulators bind to direct repeats, the 19-bp sequence might be interpreted as a loosely conserved 8-bp direct repeat separated by 3 bp. This idea was supported by the fact that the highest binding affinity was observed with a perfect 8-bp direct repeat of the sequence CCTGTGAAaatCCTGTGAA. Inspection of the other target promoters revealed sequences with some similarity to this binding motif, which might represent PhoR binding sites. The in vivo relevance of the PhoR-binding site within the phoRS promoter was supported by reporter gene studies.Phosphorus is an essential nutrient for all cells and is required, e.g., for the biosynthesis of nucleotides, DNA, and RNA, but also for the regulation of protein activity by phosphorylation of histidine, aspartate, serine, threonine, or tyrosine residues. A common phosphorus source is inorganic phosphate (P i ), and cells have developed mechanisms for the acquisition, assimilation, and storage of phosphate. When P i becomes limiting, many bacteria induce the synthesis of proteins that enable them to capture the residual P i resources more efficiently and to make alternative phosphorus sources accessible. The corresponding genes are collectively named P i starvation-inducible genes, or psi genes. The phosphate starvation response, particularly its regulation, has been most carefully studied in Escherichia coli (28) and Bacillus subtilis (12).We recently started to characterize the phosphate starvation response in Corynebacterium glutamicum (13), a gram-positive soil bacterium used industrially for the production of more than 2 million tons of amino acids per year, mainly L-glutamate and L-lysine (11). An overview of the biology, genetics, physiology, and application of C. glutamicum can be found in a recent monograph (5). Phosphorus constitutes 1.5% to 2.1% of the cell dry weight of C. glutamicum (18), part of which is present as polyphosphate (15,17,21). The phosphate star...

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Formation of volutin granules inCorynebacterium glutamicum

Cited by 56 publications

References 27 publications

Accumulation of Polyphosphate in Lactobacillus spp. and Its Involvement in Stress Resistance

Accumulation of Polyphosphate in Lactobacillus spp. and Its Involvement in Stress Resistance

Link between Phosphate Starvation and Glycogen Metabolism in Corynebacterium glutamicum , Revealed by Metabolomics

Target Genes and DNA-Binding Sites of the Response Regulator PhoR fromCorynebacterium glutamicum

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