Overproduction of heterologous mannitol 1-phosphatase : a key factor for engineering mannitol production by Lactococcus lactis

H.W. Wisselink, A.P.H.A. Moers, A.E. Mars, M.H.N. Hoefnagel, W.M. de Vos, J. Hugenholtz

Research output: Contribution to journalArticleAcademicpeer-review

39 Citations (Scopus)

Abstract

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.
Original languageEnglish
Pages (from-to)1507-1514
JournalApplied and Environmental Microbiology
Volume71
Issue number3
DOIs
Publication statusPublished - 2005

Keywords

  • controlled gene-expression
  • complete genome sequence
  • acid bacteria
  • lactate-dehydrogenase
  • lactobacillus-plantarum
  • leuconostoc
  • deficient
  • protects
  • glucose
  • mesenteroides

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