Abstract
A plasmid that expressed pyruvate carboxylase (PYC) from Rhizobium etli was introduced into Salmonella typhimurium LT2. Anaerobic fermentations of S. typhimurium with and without PYC were compared with glucose as a carbon source. The presence of PYC increased the succinate yield from glucose from 0.044 g g−1 to 0.22 g g−1, while the lactate yield decreased from 0.31 g g−1 to 0.16 g g−1. Metabolic flux calculations during the early growth phase indicate that under these growth conditions in the presence of PYC more carbon flows to oxaloacetate via pyruvate carboxylase than via phosphoenolpyruvate carboxylase. Also, under these growth and induction conditions, the presence of PYC diminished the cell growth rate from 0.34 h−1 to 0.28 h−1, the specific rate of ATP formation from 45 mmol l−1 h−1 to 27 mmol l−1 h−1, and the specific rate of glucose consumption from 17 mmol l−1 h−1 to 10 mmol l−1 h−1.
Similar content being viewed by others
References
Bunch PK, Mat-Jan F, Lee N, Clark DP (1997) The ldhA gene encoding the fermentative lactate dehydrogenase of Escherichia coli. Microbiology 143: 187-195.
Dunn MF, Encarnacion G, Araiza G, Vargas MC, Davalos A, Peralta H, Mora Y, Mora J (1996) Pyruvate carboxylase from Rhizobium etli: mutant characterizatioin, nucleotide sequence, and physiological role. J. Bacteriol. 178: 5960-5970.
Eiteman MA, Chastain MJ (1997) Optimization of the ion-exchange analysis of organic acids from fermentation. Anal. Chim. Acta 338: 69-75.
Gokarn RR, Eiteman MA, Altman E (1998) Expression of pyruvate carboxylase enhances succinate production in Escherichia coli without affecting glucose uptake. Biotechnol. Lett. 20: 795-798.
Gokarn RR, Eiteman MA, Altman E (2000) Metabolic analysis of Escherichia coli in the presence and absence of the carboxylating enzymes phophoenolpyruvate carboxylase and pruvate carboxylase. Appl. Environ. Microbiol. 66: 1844-1850.
Gottschalk G (1985) Bacterial Metabolism, 2nd edn. New York: Springer.
Liljeqvist S, Haddad D, Berzins K, Uhlén M, Ståhl S (1996) A novel expression system for Salmonella typhimurium allowing high production levels, product secretion and efficient recovery. Biochem. Biophys. Res. Commun. 218: 356-359.
Maeba P, Sanwal BD (1965) Feedback inhibition of phosphoenolpyruvate carboxylase of Salmonella. Biochem. Biophys. Res. Commun. 21: 503-508.
Maeba P, Sanwal BD (1969) Phosphoenolpyruvate carboxylase of Salmonella. Some chemical and allosteric properties. J. Biol. Chem. 13: 283-288.
Martin-Gallardo A, Fleischer E, Doyle SA, Arumugham R, Collins PL, Hildreth SW, Paradiso PR (1993) Expression of the G glycoprotein gene of human respiroatory syncytial virus in Salmonella typhimurium. J. Gen. Virol. 74: 453-458.
Payne J, Morris JG (1969) Pyruvate carboxylase in Rhodopseudomonas spheroides. J. Gen. Microbiol. 59: 97-101.
Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC (1985) Measurement of protein using bicinchoninic acid. Anal. Biochem. 150: 76-85.
Tarmy EM, Kaplan NO (1968a) Chemical characterizationi of dlactate dehydrogenase from Escherichia coli B. J. Biol. Chem. 243: 2579-2586.
Tarmy EM, Kaplan NO (1968b) Kinetic of Escherichia coli B d-lactate dehydrogenase and evidence for pyruvate-controlled change in conformation. J. Biol. Chem. 243: 2587-2596.
Theodore TS, Englesberg E (1964) Mutant of Salmonella typhimurium deficient in the carbon dixoxide-fixing enzyme phosphoenolpyruvate carboxylase. J. Bacteriol. 88: 946-955.
Tsai SP, Lee YH (1988) Application of metabolic pathway stoichiometry to statistical analysis of bioreactor measurement data. Biotechnol. Bioeng. 32: 713-715.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Xie, L., Lee, S., Hanel, B. et al. Anaerobic fermentation of Salmonella typhimurium with and without pyruvate carboxylase . Biotechnology Letters 23, 111–117 (2001). https://doi.org/10.1023/A:1010383527130
Issue Date:
DOI: https://doi.org/10.1023/A:1010383527130