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Applied and Environmental Microbiology, March 2005, p. 1507-1514, Vol. 71, No. 3
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.3.1507-1514.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Overproduction of Heterologous Mannitol 1-Phosphatase: a Key Factor for Engineering Mannitol Production by Lactococcus lactis

H. Wouter Wisselink,1,2 Antoine P. H. A. Moers,2 Astrid E. Mars,1,2 Marcel H. N. Hoefnagel,3 Willem M. de Vos,1 and Jeroen Hugenholtz1,4*

Wageningen Centre for Food Sciences (WCFS),1 Wageningen University and Research Centre—Agrotechnology and Food Innovations, Wageningen,2 NIZO Food Research, Ede,4 Centre for Biological Medicines and Medical Technology, RIVM, Bilthoven, The Netherlands3

Received 26 July 2004/ Accepted 18 October 2004

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.

* 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, March 2005, p. 1507-1514, Vol. 71, No. 3
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.3.1507-1514.2005
Copyright © 2005, 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]  


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