Metabolic Analysis of Glutamate Production by Corynebacterium glutamicum

Gourdon, Pierre; Lindley, Nic D.

doi:10.1006/mben.1999.0122

Cited by 41 publications

(19 citation statements)

References 27 publications

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“…While some of them are pathogenic to plants, animals and humans, the non-pathogenic corynebacteria, such as Corynebacterium glutamicum, are widely used in the industrial production of amino acids and nucleotides (Gourdon & Lindley, 1999;Nakayama et al, 1978).…”

Section: Introductionmentioning

confidence: 99%

Morphological changes and proteome response of Corynebacterium glutamicum to a partial depletion of FtsI

et al. 2006

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In Corynebacterium glutamicum, as in many Gram-positive bacteria, the cell division gene ftsI is located at the beginning of the dcw cluster, which comprises cell division-and cell wall-related genes. Transcriptional analysis of the cluster revealed that ftsI is transcribed as part of a polycistronic mRNA, which includes at least mraZ, mraW, ftsL, ftsI and murE, from a promoter that is located upstream of mraZ. ftsI appears also to be expressed from a minor promoter that is located in the intergenic ftsL-ftsI region. It is an essential gene in C. glutamicum, and a reduced expression of ftsI leads to the formation of larger and filamentous cells. A translational GFP-FtsI fusion protein was found to be functional and localized to the mid-cell of a growing bacterium, providing evidence of its role in cell division in C. glutamicum. This study involving proteomic analysis (using 2D SDS-PAGE) of a C. glutamicum strain that has partially depleted levels of FtsI reveals that at least 20 different proteins were overexpressed in the organism. Eight of these overexpressed proteins, which include DivIVA, were identified by MALDI-TOF. Overexpression of DivIVA was confirmed by Western blotting using anti-DivIVA antibodies, and also by fluorescence microscopy analysis of a C. glutamicum RESF1 strain expressing a chromosomal copy of a divIVA-gfp transcriptional fusion. Overexpression of DivIVA was not observed when FtsI was inhibited by cephalexin treatment or by partial depletion of FtsZ.

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Section: Introductionmentioning

confidence: 99%

Morphological changes and proteome response of Corynebacterium glutamicum to a partial depletion of FtsI

et al. 2006

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“…Some are plant and animal pathogens, and other non-pathogenic corynebacteria (Brevibacterium lactofermentum or Corynebacterium glutamicum) are used in the industrial production of amino acids and nucleotides (Gourdon & Lindley, 1999;Nakayama et al, 1978). Hermann et al (1998) identified the presence of a typical mycobacterial antigen (Antigen 84, Ag84) in C. glutamicum by protein microsequencing after two-dimensional gel electrophoresis (Hermann et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

Involvement of DivIVA in the morphology of the rod-shaped actinomycete Brevibacterium lactofermentum

et al. 2003

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In Brevibacterium lactofermentum, as in many Gram-positive bacteria, a divIVA gene is located downstream from the dcw cluster of cell-division-and cell-wall-related genes. This gene (divIVA BL ) is mostly expressed during exponential growth, and the protein encoded, DivIVA BL, bears some sequence similarity to antigen 84 (Ag84) from mycobacteria and was detected with monoclonal antibodies against Ag84. Disruption experiments using an internal fragment of the divIVA BL gene or a disrupted divIVA BL cloned in a suicide conjugative plasmid were unsuccessful, suggesting that the divIVA BL gene is needed for cell viability in Brev. lactofermentum. Transformation of Brev. lactofermentum with a multicopy plasmid containing divIVA BL drastically altered the morphology of the corynebacterial cells, which became larger and bulkier, and a GFP fusion to DivIVA BL mainly localized to the ends of corynebacterial cells. This localization pattern, together with the overproduction phenotype, suggests that DivIVA may be important in regulating the apical growth of daughter cells.

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“…However, the recent development of metabolic engineering tools, allowing the rational design of microorganisms for metabolite production (27), prompted us to evaluate an alternative process for trehalose overproduction in the gram-positive bacterium Corynebacterium glutamicum (23). C. glutamicum was chosen for three major reasons: (i) it produces, and excretes, trehalose (15); (ii) it has a metabolic control architecture simpler than that of other microorganisms, maybe as a result of its comparatively small 3,500-kb genome size (10); and (iii) it is widely used in industrial biotechnological processes (10).…”

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Overproduction of Trehalose: Heterologous Expression of Escherichia coli Trehalose-6-Phosphate Synthase and Trehalose-6-Phosphate Phosphatase in Corynebacterium glutamicum

Padilla

Krämer

Stephanopoulos

et al. 2004

Appl Environ Microbiol

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Trehalose is a disaccharide with potential applications in the biotechnology and food industries. We propose a method for industrial production of trehalose, based on improved strains of Corynebacterium glutamicum. This paper describes the heterologous expression of Escherichia coli trehalose-synthesizing enzymes trehalose-6-phosphate synthase (OtsA) and trehalose-6-phosphate phosphatase (OtsB) in C. glutamicum, as well as its impact on the trehalose biosynthetic rate and metabolic-flux distributions, during growth in a defined culture medium. The new recombinant strain showed a five-to sixfold increase in the activity of OtsAB pathway enzymes, compared to a control strain, as well as an almost fourfold increase in the trehalose excretion rate during the exponential growth phase and a twofold increase in the final titer of trehalose. The heterologous expression described resulted in a reduced specific glucose uptake rate and Krebs cycle flux, as well as reduced pentose pathway flux, a consequence of downregulated glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase. The results proved the suitability of using the heterologous expression of Ots proteins in C. glutamicum to increase the trehalose biosynthetic rate and yield and suggest critical points for further improvement of trehalose overproduction in C. glutamicum.Trehalose (1-␣-glucopyranosyl-1-␣-glucopyranoside) is a nonreducing, particularly stable disaccharide formed by two glucose moieties (37). As a compatible osmolite (22) and protein stabilizer (39), trehalose shows a wide range of potential applications in biotechnology (increased stress tolerance of important crops, stability of recombinant proteins, etc.), as well as in the food industry (37).In the past, trehalose was produced by using Saccharomyces cerevisiae (32). The high cost of this system and the promising applications of the disaccharide led to the development of a new trehalose production process based on the enzymatic biotransformation of maltodextrins (25,26). The success of the enzymatic process has limited the interest in using microorganisms as alternative sources for trehalose synthesis. However, the recent development of metabolic engineering tools, allowing the rational design of microorganisms for metabolite production (27), prompted us to evaluate an alternative process for trehalose overproduction in the gram-positive bacterium Corynebacterium glutamicum (23). C. glutamicum was chosen for three major reasons: (i) it produces, and excretes, trehalose (15); (ii) it has a metabolic control architecture simpler than that of other microorganisms, maybe as a result of its comparatively small 3,500-kb genome size (10); and (iii) it is widely used in industrial biotechnological processes (10).Three pathways for trehalose synthesis have been characterized in C. glutamicum (45), similarly to that found in Mycobacterium species (7) (Fig. 1). The first is the TreS pathway, in which trehalose is formed by maltose isomerization (7, 31).The second is the TreY-TreZ pathway, le...

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Metabolic Analysis of Glutamate Production by Corynebacterium glutamicum

Cited by 41 publications

References 27 publications

Morphological changes and proteome response of Corynebacterium glutamicum to a partial depletion of FtsI

Morphological changes and proteome response of Corynebacterium glutamicum to a partial depletion of FtsI

Involvement of DivIVA in the morphology of the rod-shaped actinomycete Brevibacterium lactofermentum

Overproduction of Trehalose: Heterologous Expression of Escherichia coli Trehalose-6-Phosphate Synthase and Trehalose-6-Phosphate Phosphatase in Corynebacterium glutamicum

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