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. 1996 Jul;62(7):2641–2643. doi: 10.1128/aem.62.7.2641-2643.1996

Genetic manipulation of the pathway for diacetyl metabolism in Lactococcus lactis.

S R Swindell 1, K H Benson 1, H G Griffin 1, P Renault 1, S D Ehrlich 1, M J Gasson 1
PMCID: PMC168044  PMID: 8779601

Abstract

Diacetyl is an important food flavor compound produced by certain strains of citrate-metabolizing lactic acid bacteria. Citrate is converted to pyruvate, from which diacetyl is produced via intermediate alpha-acetolactate. This paper reports the cloning and analysis of the gene (aldB) encoding alpha-acetolactate decarboxylase from Lactococcus lactis MG1363. Deletion of the MG1363 chromosomal aldB gene was achieved by double crossover homologous recombination. The mutant strain was found to produce diacetyl; alpha-acetolactate decarboxylase activity was eliminated. Overexpression of the cloned ilvBN genes (encoding an alpha-acetolactate synthase) in the aldB deletion strain produced even higher levels of alpha-acetolactate, acetoin, and diacetyl.

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Selected References

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  1. Bassit N., Boquien C. Y., Picque D., Corrieu G. Effect of Initial Oxygen Concentration on Diacetyl and Acetoin Production by Lactococcus lactis subsp. lactis biovar diacetylactis. Appl Environ Microbiol. 1993 Jun;59(6):1893–1897. doi: 10.1128/aem.59.6.1893-1897.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  3. Gasson M. J. Plasmid complements of Streptococcus lactis NCDO 712 and other lactic streptococci after protoplast-induced curing. J Bacteriol. 1983 Apr;154(1):1–9. doi: 10.1128/jb.154.1.1-9.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Godon J. J., Chopin M. C., Ehrlich S. D. Branched-chain amino acid biosynthesis genes in Lactococcus lactis subsp. lactis. J Bacteriol. 1992 Oct;174(20):6580–6589. doi: 10.1128/jb.174.20.6580-6589.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Godon J. J., Delorme C., Bardowski J., Chopin M. C., Ehrlich S. D., Renault P. Gene inactivation in Lactococcus lactis: branched-chain amino acid biosynthesis. J Bacteriol. 1993 Jul;175(14):4383–4390. doi: 10.1128/jb.175.14.4383-4390.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Goupil N., Corthier G., Ehrlich S. D., Renault P. Imbalance of leucine flux in Lactococcus lactis and its use for the isolation of diacetyl-overproducing strains. Appl Environ Microbiol. 1996 Jul;62(7):2636–2640. doi: 10.1128/aem.62.7.2636-2640.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Griffin H. G., I'Anson K. J., Gasson M. J. Rapid isolation of genes from bacterial lambda libraries by direct polymerase chain reaction screening. FEMS Microbiol Lett. 1993 Aug 15;112(1):49–53. doi: 10.1111/j.1574-6968.1993.tb06422.x. [DOI] [PubMed] [Google Scholar]
  8. Phalip V., Monnet C., Schmitt P., Renault P., Godon J. J., Diviès C. Purification and properties of the alpha-acetolactate decarboxylase from Lactococcus lactis subsp. lactis NCDO 2118. FEBS Lett. 1994 Aug 29;351(1):95–99. doi: 10.1016/0014-5793(94)00820-5. [DOI] [PubMed] [Google Scholar]
  9. Renault P., Godon J. J., Goupil N., Delorme C., Corthier G., Ehrlich S. D. Metabolic operons in Lactococci. Dev Biol Stand. 1995;85:431–441. [PubMed] [Google Scholar]
  10. Snoep J. L., Teixeira de Mattos M. J., Starrenburg M. J., Hugenholtz J. Isolation, characterization, and physiological role of the pyruvate dehydrogenase complex and alpha-acetolactate synthase of Lactococcus lactis subsp. lactis bv. diacetylactis. J Bacteriol. 1992 Jul;174(14):4838–4841. doi: 10.1128/jb.174.14.4838-4841.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Starrenburg M. J., Hugenholtz J. Citrate Fermentation by Lactococcus and Leuconostoc spp. Appl Environ Microbiol. 1991 Dec;57(12):3535–3540. doi: 10.1128/aem.57.12.3535-3540.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. van der Vossen J. M., van der Lelie D., Venema G. Isolation and characterization of Streptococcus cremoris Wg2-specific promoters. Appl Environ Microbiol. 1987 Oct;53(10):2452–2457. doi: 10.1128/aem.53.10.2452-2457.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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