This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Platteeuw, C. Articles by de Vos, W. M. Search for Related Content PubMed PubMed Citation Articles by Platteeuw, C. Articles by de Vos, W. M. Agricola Articles by Platteeuw, C. Articles by de Vos, W. M.

 Previous Article  |  Next Article 

Appl. Environ. Microbiol., 11 1995, 3967-3971, Vol 61, No. 11
Copyright © 1995, American Society for Microbiology

Metabolic engineering of Lactococcus lactis: influence of the overproduction of alpha-acetolactate synthase in strains deficient in lactate dehydrogenase as a function of culture conditions

C Platteeuw, J Hugenholtz, M Starrenburg, I van Alen-Boerrigter and WM de Vos
Department of Biophysical Chemistry, NIZO, Ede, The Netherlands.

The als gene for alpha-acetolactate synthase of Lactococcus lactis MG1363 was cloned on a multicopy plasmid under the control of the inducible L. lactis lacA promoter. More than a hundredfold overproduction of alpha-acetolactate synthase was obtained in L. lactis under inducing conditions as compared with that of the host strain, which contained a single chromosomal copy of the als gene. The effect of alpha-acetolactate synthase overproduction on the formation of end products in various L. lactis strains was studied under different fermentation conditions. Under aerobic conditions and with an initial pH of 6.0, overexpression of the als gene resulted in significant acetoin production that amounted to more than one-third of the pyruvate converted. However, the effect of the alpha-acetolactate synthase overproduction was even more pronounced in the lactate dehydrogenase- deficient strain L. lactis NZ2700. Anaerobic cultivation of this strain resulted in a doubling of the butanediol formation of up to 40% of the converted pyruvate. When cultivated aerobically at an initial pH of 6.8, overexpression of the als gene in L. lactis NZ2700 resulted in the conversion of more than 60% of the pyruvate into acetoin, while no butanediol was formed. Moreover, at an initial pH of 6.0, similar amounts of acetoin were obtained, but in addition approximately 20% of the pyruvate was converted into butanediol. These metabolic engineering studies indicate that more than 80% of the lactose can be converted via the activity of the overproduced alpha-acetolactate synthase in L. lactis.

This article has been cited by other articles:

• Larsen, N., Boye, M., Siegumfeldt, H., Jakobsen, M. (2006). Differential Expression of Proteins and Genes in the Lag Phase of Lactococcus lactis subsp. lactis Grown in Synthetic Medium and Reconstituted Skim Milk. Appl. Environ. Microbiol. 72: 1173-1179 [Abstract] [Full Text]  
• Bongers, R. S., Hoefnagel, M. H. N., Kleerebezem, M. (2005). High-Level Acetaldehyde Production in Lactococcus lactis by Metabolic Engineering. Appl. Environ. Microbiol. 71: 1109-1113 [Abstract] [Full Text]  
• Vido, K., le Bars, D., Mistou, M.-Y., Anglade, P., Gruss, A., Gaudu, P. (2004). Proteome Analyses of Heme-Dependent Respiration in Lactococcus lactis: Involvement of the Proteolytic System. J. Bacteriol. 186: 1648-1657 [Abstract] [Full Text]  
• Gaspar, P., Neves, A. R., Ramos, A., Gasson, M. J., Shearman, C. A., Santos, H. (2004). Engineering Lactococcus lactis for Production of Mannitol: High Yields from Food-Grade Strains Deficient in Lactate Dehydrogenase and the Mannitol Transport System. Appl. Environ. Microbiol. 70: 1466-1474 [Abstract] [Full Text]  
• Bongers, R. S., Hoefnagel, M. H. N., Starrenburg, M. J. C., Siemerink, M. A. J., Arends, J. G. A., Hugenholtz, J., Kleerebezem, M. (2003). IS981-Mediated Adaptive Evolution Recovers Lactate Production by ldhB Transcription Activation in a Lactate Dehydrogenase-Deficient Strain of Lactococcus lactis. J. Bacteriol. 185: 4499-4507 [Abstract] [Full Text]  
• Koebmann, B. J., Solem, C., Pedersen, M. B., Nilsson, D., Jensen, P. R. (2002). Expression of Genes Encoding F1-ATPase Results in Uncoupling of Glycolysis from Biomass Production in Lactococcus lactis. Appl. Environ. Microbiol. 68: 4274-4282 [Abstract] [Full Text]  
• Hoefnagel, M. H. N., Starrenburg, M. J. C., Martens, D. E., Hugenholtz, J., Kleerebezem, M., Van Swam, I. I., Bongers, R., Westerhoff, H. V., Snoep, J. L. (2002). Metabolic engineering of lactic acid bacteria, the combined approach: kinetic modelling, metabolic control and experimental analysis. Microbiology 148: 1003-1013 [Abstract] [Full Text]  
• Gosalbes, M. J., Esteban, C. D., Galán, J. L., Pérez-Martínez, G. (2000). Integrative Food-Grade Expression System Based on the Lactose Regulon of Lactobacillus casei. Appl. Environ. Microbiol. 66: 4822-4828 [Abstract] [Full Text]  
• Hugenholtz, J., Kleerebezem, M., Starrenburg, M., Delcour, J., de Vos, W., Hols, P. (2000). Lactococcus lactis as a Cell Factory for High-Level Diacetyl Production. Appl. Environ. Microbiol. 66: 4112-4114 [Abstract] [Full Text]  
• Hols, P., Ramos, A., Hugenholtz, J., Delcour, J., de Vos, W. M., Santos, H., Kleerebezem, M. (1999). Acetate Utilization in Lactococcus lactis Deficient in Lactate Dehydrogenase: a Rescue Pathway for Maintaining Redox Balance. J. Bacteriol. 181: 5521-5526 [Abstract] [Full Text]  
• Curic, M., Stuer-Lauridsen, B., Renault, P., Nilsson, D. (1999). A General Method for Selection of alpha -Acetolactate Decarboxylase-Deficient Lactococcus lactis Mutants To Improve Diacetyl Formation. Appl. Environ. Microbiol. 65: 1202-1206 [Abstract] [Full Text]  
• Lopez de Felipe, F., Kleerebezem, M., de Vos, W. M., Hugenholtz, J. (1998). Cofactor Engineering: a Novel Approach to Metabolic Engineering in Lactococcus lactis by Controlled Expression of NADH Oxidase. J. Bacteriol. 180: 3804-3808 [Abstract] [Full Text]