Overproduction of heterologous mannitol 1-phosphatase : a key factor for engineering mannitol production by Lactococcus lactis
Wisselink, H.W. ; Moers, A.P.H.A. ; Mars, A.E. ; Hoefnagel, M.H.N. ; Vos, W.M. de; Hugenholtz, J. - \ 2005
Applied and Environmental Microbiology 71 (2005)3. - ISSN 0099-2240 - p. 1507 - 1514.
controlled gene-expression - complete genome sequence - acid bacteria - lactate-dehydrogenase - lactobacillus-plantarum - leuconostoc - deficient - protects - glucose - mesenteroides
To achieve high mannitol production by Lactococcus lactis, the mannitol 1-phosphatase gene of Eimeria tenella and the mannitol 1-phosphate dehydrogenase gene mtlD of Lactobacillus plantarum were cloned in the nisin-dependent L. lactis NICE overexpression system. As predicted by a kinetic L. lactis glycolysis model, increase in mannitol 1-phosphate dehydrogenase and mannitol 1-phosphatase activities resulted in increased mannitol production. Overexpression of both genes in growing cells resulted in glucose-mannitol conversions of 11, 21, and 27% by the L. lactis parental strain, a strain with reduced phosphofructokinase activity, and a lactate dehydrogenase-deficient strain, respectively. Improved induction conditions and increased substrate concentrations resulted in an even higher glucose-to-mannitol conversion of 50% by the lactate dehydrogenase-deficient L. lactis strain, close to the theoretical mannitol yield of 67%. Moreover, a clear correlation between mannitol 1-phosphatase activity and mannitol production was shown, demonstrating the usefulness of this metabolic engineering approach.
Metabolic Engineering of Mannitol Production in Lactococcus lactis: Influence of Overexpression of Mannitol 1-Phosphate Dehydrogenase in Different Genetic Backgrounds
Wisselink, H.W. ; Mars, A.E. ; Meer, P. van der; Eggink, G. ; Hugenholtz, J. - \ 2004
Applied and Environmental Microbiology 70 (2004)7. - ISSN 0099-2240 - p. 4286 - 4292.
complete genome sequence - lactate-dehydrogenase - lactobacillus-plantarum - acid bacteria - phosphotransferase system - expression systems - protects - nisin - phosphorylation - activation
To obtain a mannitol-producing Lactococcus lactis strain, the mannitol 1-phosphate dehydrogenase gene (mtlD) from Lactobacillus plantarum was overexpressed in a wild-type strain, a lactate dehydrogenase(LDH)-deficient strain, and a strain with reduced phosphofructokinase activity. High-performance liquid chromatography and 13C nuclear magnetic resonance analysis revealed that small amounts (