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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.

Metabolic Engineering of Mannitol Production in Lactococcus lactis: Influence of Overexpression of Mannitol 1-Phosphate Dehydrogenase in Different Genetic Backgrounds

H. Wouter Wisselink,1,2 Astrid E. Mars,1,2 Pieter van der Meer,2 Gerrit Eggink,1,2 and Jeroen Hugenholtz1,3*

Wageningen Centre for Food Sciences,1 Wageningen University and Research Centre—Agrotechnology 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.

* Corresponding author. Mailing address: NIZO Food Research, Kernhemseweg 2, 6718 ZB Ede, The Netherlands. Phone: (31) 318 659511. Fax: (31) 317 650400. E-mail: Jeroen.Hugenholtz{at}nizo.nl.

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.



This article has been cited by other articles:

  • Ladero, V., Ramos, A., Wiersma, A., Goffin, P., Schanck, A., Kleerebezem, M., Hugenholtz, J., Smid, E. J., Hols, P. (2007). High-Level Production of the Low-Calorie Sugar Sorbitol by Lactobacillus plantarum through Metabolic Engineering. Appl. Environ. Microbiol. 73: 1864-1872 [Abstract] [Full Text]  
  • Wisselink, H. W., Moers, A. P. H. A., Mars, A. E., Hoefnagel, M. H. N., de Vos, W. M., Hugenholtz, J. (2005). Overproduction of Heterologous Mannitol 1-Phosphatase: a Key Factor for Engineering Mannitol Production by Lactococcus lactis. Appl. Environ. Microbiol. 71: 1507-1514 [Abstract] [Full Text]  


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