J. Bacteriol., Nov 1996, 6209-6215, Vol 178, No. 21
Copyright © 1996, American Society for Microbiology

The citrate metabolic pathway in Leuconostoc mesenteroides: expression, amino acid synthesis, and alpha-ketocarboxylate transport

C Marty-Teysset, JS Lolkema, P Schmitt, C Divies and WN Konings
Department of Microbiology, Groningen Biotechnology and Biomolecular Sciences Institute, University of Groningen, The Netherlands.

Citrate metabolism in Leuconostoc mesenteroides subspecies mesenteroides is associated with the generation of a proton motive force by a secondary mechanism (C. Marty-Teysset, C. Posthuma, J. S. Lolkema, P. Schmitt, C. Divies, and W. N. Konings, J. Bacteriol. 178:2178-2185, 1996). The pathway consists of four steps: (i) uptake of citrate, (ii) splitting of citrate into acetate and oxaloacetate, (iii) pyruvate formation by decarboxylation of oxaloacetate, and (iv) reduction of pyruvate to lactate. Studies of citrate uptake and metabolism in resting cells of L. mesenteroides grown in the presence or absence of citrate show that the citrate transporter CitP and citrate lyase are constitutively expressed. On the other hand, oxaloacetate decarboxylase is under stringent control of the citrate in the medium and is not expressed in its absence, thereby blocking the pathway at the level of oxaloacetate. Under those conditions, the pathway is completely directed towards the formation of aspartate, which is formed from oxaloacetate by transaminase activity. The data indicate a role for citrate metabolism in amino acid biosynthesis. Internalized radiolabeled aspartate produced from citrate metabolism could be chased from the cells by addition of the amino acid precursors oxaloacetate, pyruvate, alpha-ketoglutarate, and alpha-ketoisocaproate to the cells, indicating a broad specificity of the transamination reaction. The alpha-ketocarboxylates are readily transported across the cytoplasmic membrane. alpha-Ketoglutarate uptake in resting cells of L. mesenteroides was dependent upon the presence of an energy source and was inhibited by inhibition of the proton motive force generating F(0)F(1) ATPase and by selective dissipation of the membrane potential and the transmembrane pH gradient. It is concluded that in L. mesenteroides alpha-ketoglutarate is transported via a secondary transporter that may be a general alpha-ketocarboxylate carrier.

This article has been cited by other articles:

• Sobczak, I., Lolkema, J. S. (2005). The 2-Hydroxycarboxylate Transporter Family: Physiology, Structure, and Mechanism. Microbiol. Mol. Biol. Rev. 69: 665-695 [Abstract] [Full Text]  
• Magni, C., de Mendoza, D., Konings, W. N., Lolkema, J. S. (1999). Mechanism of Citrate Metabolism in Lactococcus lactis: Resistance against Lactate Toxicity at Low pH. J. Bacteriol. 181: 1451-1457 [Abstract] [Full Text]  
• Bandell, M., Lhotte, M. E., Marty-Teysset, C., Veyrat, A., Prévost, H., Dartois, V., Diviès, C., Konings, W. N., Lolkema, J. S. (1998). Mechanism of the Citrate Transporters in Carbohydrate and Citrate Cometabolism in Lactococcus and Leuconostoc Species. Appl. Environ. Microbiol. 64: 1594-1600 [Abstract] [Full Text]  
• Wang, H., Yu, W., Coolbear, T., O'Sullivan, D., McKay, L. L. (1998). A Deficiency in Aspartate Biosynthesis in Lactococcus lactis subsp. lactis C2 Causes Slow Milk Coagulation. Appl. Environ. Microbiol. 64: 1673-1679 [Abstract] [Full Text]  
• Bandell, M., Ansanay, V., Rachidi, N., Dequin, S., Lolkema, J. S. (1997). Membrane Potential-generating Malate (MleP) and Citrate (CitP) Transporters of Lactic Acid Bacteria Are Homologous Proteins. SUBSTRATE SPECIFICITY OF THE 2-HYDROXYCARBOXYLATE TRANSPORTER FAMILY. J. Biol. Chem. 272: 18140-18146 [Abstract] [Full Text]