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Applied and Environmental Microbiology, July 2004, p. 4286-4292, Vol. 70, No. 7
0099-2240/04/$08.00+0 DOI: 10.1128/AEM.70.7.4286-4292.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.
Wageningen Centre for Food Sciences,1 Wageningen University and Research CentreAgrotechnology and Food Innovations, 6700 AA Wageningen,2 NIZO Food Research, 6710 BA Ede, The Netherlands3
Received 22 November 2003/ Accepted 12 March 2004
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 (<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.
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