Characterization of 3-phosphoglycerate kinase from *Corynebacterium glutamicum* and its impact on amino acid production

Komati Reddy G, Wendisch VF (2014)
BMC Microbiology 14(54).

Download
OA
Journal Article | Published | English
Abstract
Background Corynebacterium glutamicum cg1790/pgk encodes an enzyme active as a 3-phosphoglycerate kinase (PGK) (EC 2.7.2.3) catalyzing phosphoryl transfer from 1,3-biphosphoglycerate (bPG) to ADP to yield 3-phosphoglycerate (3-PG) and ATP in substrate chain phosphorylation. Results C. glutamicum 3-phosphoglycerate kinase was purified to homogeneity from the soluble fraction of recombinant E. coli. PGKHis was found to be active as a homodimer with molecular weight of 104 kDa. The enzyme preferred conditions of pH 7.0 to 7.4 and required Mg2+ for its activity. PGKHis is thermo labile and it has shown maximal activity at 50–65°C. The maximal activity of PGKHis was estimated to be 220 and 150 U mg-1 with KM values of 0.26 and 0.11 mM for 3-phosphoglycerate and ATP, respectively. A 3-phosphoglycerate kinase negative C. glutamicum strain ∆pgk was constructed and shown to lack the ability to grow under glycolytic or gluconeogenic conditions unless PGK was expressed from a plasmid to restore growth. When pgk was overexpressed in L-arginine and L-ornithine production strains the production increased by 8% and by 17.5%, respectively. Conclusion Unlike many bacterial PGKs, C. glutamicum PGK is active as a homodimer. PGK is essential for growth of C. glutamicum with carbon sources requiring glycolysis and gluconeogenesis. Competitive inhibition by ADP reveals the critical role of PGK in gluconeogenesis by energy charge. Pgk overexpression improved the productivity in L-arginine and L-ornithine production strains.
Publishing Year
ISSN
PUB-ID

Cite this

Komati Reddy G, Wendisch VF. Characterization of 3-phosphoglycerate kinase from *Corynebacterium glutamicum* and its impact on amino acid production. BMC Microbiology. 2014;14(54).
Komati Reddy, G., & Wendisch, V. F. (2014). Characterization of 3-phosphoglycerate kinase from *Corynebacterium glutamicum* and its impact on amino acid production. BMC Microbiology, 14(54).
Komati Reddy, G., and Wendisch, V. F. (2014). Characterization of 3-phosphoglycerate kinase from *Corynebacterium glutamicum* and its impact on amino acid production. BMC Microbiology 14.
Komati Reddy, G., & Wendisch, V.F., 2014. Characterization of 3-phosphoglycerate kinase from *Corynebacterium glutamicum* and its impact on amino acid production. BMC Microbiology, 14(54).
G. Komati Reddy and V.F. Wendisch, “Characterization of 3-phosphoglycerate kinase from *Corynebacterium glutamicum* and its impact on amino acid production”, BMC Microbiology, vol. 14, 2014.
Komati Reddy, G., Wendisch, V.F.: Characterization of 3-phosphoglycerate kinase from *Corynebacterium glutamicum* and its impact on amino acid production. BMC Microbiology. 14, (2014).
Komati Reddy, Gajendar, and Wendisch, Volker F. “Characterization of 3-phosphoglycerate kinase from *Corynebacterium glutamicum* and its impact on amino acid production”. BMC Microbiology 14.54 (2014).
Main File(s)
Access Level
OA Open Access
Last Uploaded
2015-12-11T14:09:45Z

This data publication is cited in the following publications:
This publication cites the following data publications:

61 References

Data provided by Europe PubMed Central.

Taxonomical studies on glutamic acid producing bacteria
AUTHOR UNKNOWN, 1967
Putrescine production by engineered Corynebacterium glutamicum.
Schneider J, Wendisch VF., Appl. Microbiol. Biotechnol. 88(4), 2010
PMID: 20661733

AUTHOR UNKNOWN, 1989
Studies on transformation of Escherichia coli with plasmids.
Hanahan D., J. Mol. Biol. 166(4), 1983
PMID: 6345791

AUTHOR UNKNOWN, 2005
Characterization of a Corynebacterium glutamicum lactate utilization operon induced during temperature-triggered glutamate production.
Stansen C, Uy D, Delaunay S, Eggeling L, Goergen JL, Wendisch VF., Appl. Environ. Microbiol. 71(10), 2005
PMID: 16204505
Pyruvate carboxylase is a major bottleneck for glutamate and lysine production by Corynebacterium glutamicum.
Peters-Wendisch PG, Schiel B, Wendisch VF, Katsoulidis E, Mockel B, Sahm H, Eikmanns BJ., J. Mol. Microbiol. Biotechnol. 3(2), 2001
PMID: 11321586

Export

0 Marked Publications

Open Data PUB

Web of Science

View record in Web of Science®

Sources

PMID: 24593686
PubMed | Europe PMC

Search this title in

Google Scholar