To explore the factors controlling metabolite formation under aeration in Lactococcus lactis, metabolic patterns, enzymatic activities, and transcriptional profiles of genes involved in the aerobic pathway for acetate anabolism were compared between a parental L. lactis strain and its NADH-oxidase-overproducer derivative. Deregulated catabolite repression mutans in the ccpA or pstH genes, encoding CcpA or its co-activator HPr, respectively, were compared with a parental strain, as well. Although the NADH-oxidase activity was derepressed in ccpA, but not in the pstH background, a mixed fermentation was displayed by either mutant, with a higher acetate production in the pstH variant. Moreover, transcription of genes encoding phosphotransacetylase and acetate kinase were derepressed, and the corresponding enzymatic activities increased, in both catabolite repression mutants. These results and the dependence on carbon source for acetate production in the NADH-oxidase-overproducer support the conclusion that catabolite repression, rather than NADH oxidation, plays a critical role to control acetate production. Furthermore, fructose 1,6-bisphosphate influenced the in vitro phosphotransacetylase and acetate kinase activities, while the former was sensitive to diacetyl. Our study strongly supports the model that, under aerobic conditions, the homolactic fermentation in L. lactis MG1363 is maintained by CcpA-mediated repression of mixed acid fermentation.
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We thank Dr J. Deutscher for providing us the strain pstH1 mutant (LIG101 strain) and M.-A. Petit for helpful discussion. F.L.F would like to acknowledge grant no. PR2004-0289 from the Secretaría de Estado de Educación y Universidades of Spain to promote the mobility of senior researchers.
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Lopez de Felipe, F., Gaudu, P. Multiple control of the acetate pathway in Lactococcus lactis under aeration by catabolite repression and metabolites. Appl Microbiol Biotechnol 82, 1115–1122 (2009). https://doi.org/10.1007/s00253-009-1897-8
- Lactococcus lactis
- Aerobic conditions
- Catabolite repression