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Using Lactococcus lactis for glutathione overproduction

  • Applied Genetics and Molecular Biotechnology
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Abstract

Glutathione and γ-glutamylcysteine were produced in Lactococcus lactis using a controlled expression system and the genes gshA and gshB from Escherichia coli encoding the enzymes γ-glutamylcysteine synthetase and glutathione synthetase. High levels of γ-glutamylcysteine were found in strains growing on chemically defined medium and expressing either gshA alone or both gshA and gshB. As anticipated, glutathione was found in a strain expressing gshA and gshB. The level of glutathione production could be increased by addition of the precursor amino acid cysteine to the medium. The addition of cysteine led to an increased activity of glutathione synthetase, which is remarkable because the amino acid is not a substrate of this enzyme. The final intracellular glutathione concentration attained was 358 nmol mg−1 protein, which is the highest concentration reported for a bacterium, demonstrating the suitability of engineered L. lactis for fine-chemical production and as a model for studies of the impact of glutathione on flavour formation and other properties of food.

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Acknowledgements

The authors want to thank Mr. Jan van Riel, for his assistance in HPLC analysis. This project was partly supported by National Science Foundation of China (Contract No. 30300009).

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Authors and Affiliations

  1. Wageningen Centre for Food Sciences, NIZO food research, P.O. Box 20, 6710, Ede, The Netherlands

    Yin Li, Jeroen Hugenholtz, Wilbert Sybesma & Douwe Molenaar

  2. The Key Laboratory of Industrial Biotechnology, Ministry of Education, Southern Yangtze University, Huihe Road 170, Wuxi, 214036, People’s Republic of China

    Yin Li

  3. Laboratory of Food Microbiology, Wageningen University, 6703, Wageningen, The Netherlands

    Tjakko Abee

Authors
  1. Yin Li
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  2. Jeroen Hugenholtz
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  3. Wilbert Sybesma
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  4. Tjakko Abee
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  5. Douwe Molenaar
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Correspondence to Douwe Molenaar.

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Li, Y., Hugenholtz, J., Sybesma, W. et al. Using Lactococcus lactis for glutathione overproduction. Appl Microbiol Biotechnol 67, 83–90 (2005). https://doi.org/10.1007/s00253-004-1762-8

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  • DOI: https://doi.org/10.1007/s00253-004-1762-8

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