Metabolic engineering of mannitol production in Lactococcus lactis: influence of overexpression of mannitol 1-phosphate dehydrogenase in different genetic backgrounds.
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 (13)C nuclear magnetic resonance analysis revealed that small amounts (<1%) of mannitol were formed by growing cells of mtlD-overexpressing LDH-deficient and phosphofructokinase-reduced strains, whereas resting cells of the LDH-deficient transformant converted 25% of glucose into mannitol. Moreover, the formed mannitol was not reutilized upon glucose depletion. Of the metabolic-engineering strategies investigated in this work, mtlD-overexpressing LDH-deficient L. lactis seemed to be the most promising strain for mannitol production.
Wisselink
H Wouter
HW
Wageningen Centre for Food Sciences, Wageningen University and Research Centre-Agrotechnology and Food Innovations, Ede, The Netherlands.
Mars
Astrid E
AE
van der Meer
Pieter
P
Eggink
Gerrit
G
Hugenholtz
Jeroen
J
D005818:Genetic Engineering; D005947:Glucose / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000172:deficiency; D013294:Lactococcus lactis / Q000378:metabolism*; D008353:Mannitol / Q000378:metabolism*; D025221:Phosphofructokinases / Q000378:metabolism; D013401:Sugar Alcohol Dehydrogenases / Q000235:genetics / Q000502:physiology*
D008353:Mannitol; D013401:Sugar Alcohol Dehydrogenases; C021544:mannitol-1-phosphate dehydrogenase; D007770:L-Lactate Dehydrogenase; D025221:Phosphofructokinases; D005947:Glucose