title: Overproduction of heterologous mannitol 1-phosphatase: a key factor for engineering mannitol production by Lactococcus lactis.
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.
authors.affiliation: Wageningen Centre for Food Sciences, The Netherlands.
authors.forename: H Wouter
authors.initials: HW
authors.lastname: Wisselink
authors.forename: Antoine P H A
authors.initials: AP
authors.lastname: Moers
authors.forename: Astrid E
authors.initials: AE
authors.lastname: Mars
authors.forename: Marcel H N
authors.initials: MH
authors.lastname: Hoefnagel
authors.forename: Willem M
authors.initials: WM
authors.lastname: de Vos
authors.forename: Jeroen
authors.initials: J
authors.lastname: Hugenholtz
mesh_terms: D000818:Animals; D001483:Base Sequence; D016786:Eimeria tenella / Q000201:enzymology / Q000235:genetics; D005798:Genes, Bacterial; D017125:Genes, Protozoan; D005818:Genetic Engineering; D048191:Lactiplantibacillus plantarum / Q000201:enzymology / Q000235:genetics; D013294:Lactococcus lactis / Q000235:genetics / Q000378:metabolism*; D008353:Mannitol / Q000378:metabolism*; D009561:Nisin / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000096:biosynthesis* / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013401:Sugar Alcohol Dehydrogenases / Q000096:biosynthesis / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D009561:Nisin; D008353:Mannitol; D013401:Sugar Alcohol Dehydrogenases; C021544:mannitol-1-phosphate dehydrogenase; D010744:Phosphoric Monoester Hydrolases; C021543:mannitol-1-phosphatase