Effect of Pyruvate Carboxylase Overexpression on the Physiology of Corynebacterium Glutamicum

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 2002 Oct 31

PMID 12406733

Citations 14

Authors

Mattheos A G Koffas

Gyoo Yeol Jung

Juan C Aon

Gregory Stephanopoulos

Affiliations

Soon will be listed here.

Abstract

Pyruvate carboxylase was recently sequenced in Corynebacterium glutamicum and shown to play an important role of anaplerosis in the central carbon metabolism and amino acid synthesis of these bacteria. In this study we investigate the effect of the overexpression of the gene for pyruvate carboxylase (pyc) on the physiology of C. glutamicum ATCC 21253 and ATCC 21799 grown on defined media with two different carbon sources, glucose and lactate. In general, the physiological effects of pyc overexpression in Corynebacteria depend on the genetic background of the particular strain studied and are determined to a large extent by the interplay between pyruvate carboxylase and aspartate kinase activities. If the pyruvate carboxylase activity is not properly matched by the aspartate kinase activity, pyc overexpression results in growth enhancement instead of greater lysine production, despite its central role in anaplerosis and aspartic acid biosynthesis. Aspartate kinase regulation by lysine and threonine, pyruvate carboxylase inhibition by aspartate (shown in this study using permeabilized cells), as well as well-established activation of pyruvate carboxylase by lactate and acetyl coenzyme A are the key factors in determining the effect of pyc overexpression on Corynebacteria physiology.

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References

Eikmanns B . Identification, sequence analysis, and expression of a Corynebacterium glutamicum gene cluster encoding the three glycolytic enzymes glyceraldehyde-3-phosphate dehydrogenase, 3-phosphoglycerate kinase, and triosephosphate isomerase. J Bacteriol. 1992; 174(19):6076-86. PMC: 207673. DOI: 10.1128/jb.174.19.6076-6086.1992. View

Stephanopoulos G, Vallino J . Network rigidity and metabolic engineering in metabolite overproduction. Science. 1991; 252(5013):1675-81. DOI: 10.1126/science.1904627. View

Cazzulo J, Sundaram T, Kornberg H . Regulation of pyruvate carboxylase formation from the apo-enzyme and biotin in a thermophilic bacillus. Nature. 1969; 223(5211):1137-8. DOI: 10.1038/2231137a0. View

Hartman R . Carbon dioxide fixation by extracts of Streptococcus faecalis var. liquefaciens. J Bacteriol. 1970; 102(2):341-6. PMC: 247556. DOI: 10.1128/jb.102.2.341-346.1970. View

White P . Activities of anaplerotic enzymes and acetyl coenzyme A carboxylase in biotin-deficient Bacillus megaterium. J Gen Microbiol. 1977; 100(1):203-6. DOI: 10.1099/00221287-100-1-203. View

Hanahan D . Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983; 166(4):557-80. DOI: 10.1016/s0022-2836(83)80284-8. View

Taylor B, Routman S, UTTER M . The control of the synthesis of pyruvate carboxylase in Pseudomonas citronellolis. Evience from double labeling studies. J Biol Chem. 1975; 250(6):2376-82. View

Yakunin A, Hallenbeck P . Regulation of synthesis of pyruvate carboxylase in the photosynthetic bacterium Rhodobacter capsulatus. J Bacteriol. 1997; 179(5):1460-8. PMC: 178853. DOI: 10.1128/jb.179.5.1460-1468.1997. View

Phibbs Jr P, Feary T, Blevins W . Pyruvate carboxylase deficiency in pleiotropic carbohydrate-negative mutant strains of Pseudomonas aeruginosa. J Bacteriol. 1974; 118(3):999-1009. PMC: 246850. DOI: 10.1128/jb.118.3.999-1009.1974. View

10.

Menendez J, GANCEDO C . Regulatory regions in the promoters of the Saccharomyces cerevisiae PYC1 and PYC2 genes encoding isoenzymes of pyruvate carboxylase. FEMS Microbiol Lett. 1998; 164(2):345-52. DOI: 10.1111/j.1574-6968.1998.tb13108.x. View

11.

Lachica V, Hartman P . Inhibition of CO2 fixation in group D streptococci. Can J Microbiol. 1969; 15(1):57-60. DOI: 10.1139/m69-009. View

12.

ORegan M, Thierbach G, Bachmann B, Villeval D, Lepage P, Viret J . Cloning and nucleotide sequence of the phosphoenolpyruvate carboxylase-coding gene of Corynebacterium glutamicum ATCC13032. Gene. 1989; 77(2):237-51. DOI: 10.1016/0378-1119(89)90072-3. View

13.

Sundaram T, Cazzulo J, Kornberg H . Synthesis of pyruvate carboxylase from its apoenzyme and (+)-biotin in Bacillus stearothermophilus. Mechanism and control of the reaction. Biochem J. 1971; 122(5):663-9. PMC: 1176834. DOI: 10.1042/bj1220663. View

14.

Hillier A, Jago G . The metabolism of [14C]bicarbonate by Streptococcus lactis: the fixation of [14C]bicarbonate by pyruvate carboxylase. J Dairy Res. 1978; 45(3):433-44. DOI: 10.1017/s0022029900016654. View

15.

Messerotti L, Radford A, Hodgson A . Nucleotide sequence of the replication region from the Mycobacterium-Escherichia coli shuttle vector pEP2. Gene. 1990; 96(1):147-8. DOI: 10.1016/0378-1119(90)90356-v. View

16.

Cremer J, Eggeling L, Sahm H . Control of the Lysine Biosynthesis Sequence in Corynebacterium glutamicum as Analyzed by Overexpression of the Individual Corresponding Genes. Appl Environ Microbiol. 1991; 57(6):1746-1752. PMC: 183462. DOI: 10.1128/aem.57.6.1746-1752.1991. View

17.

Peters-Wendisch P, Wendisch V, Paul S, Eikmanns B, Sahm H . Pyruvate carboxylase as an anaplerotic enzyme in . Microbiology (Reading). 2021; 143(4):1095-1103. DOI: 10.1099/00221287-143-4-1095. View

18.

Scrutton M, Taylor B . Isolation and characterization of pyruvate carboxylase from Azotobacter vinelandii OP. Arch Biochem Biophys. 1974; 164(2):641-54. DOI: 10.1016/0003-9861(74)90076-9. View

19.

Kiss R, Stephanopoulos G . Metabolic characterization of a L-lysine-producing strain by continuous culture. Biotechnol Bioeng. 1992; 39(5):565-74. DOI: 10.1002/bit.260390512. View

20.

Brewster N, Val D, Walker M, Wallace J . Regulation of pyruvate carboxylase isozyme (PYC1, PYC2) gene expression in Saccharomyces cerevisiae during fermentative and nonfermentative growth. Arch Biochem Biophys. 1994; 311(1):62-71. DOI: 10.1006/abbi.1994.1209. View