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Extracellular glutathione fermentation using engineered Saccharomyces cerevisiae expressing a novel glutathione exporter

  • Applied microbial and cell physiology
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Abstract

A novel extracellular glutathione fermentation method using engineered Saccharomyces cerevisiae was developed by following three steps. First, a platform host strain lacking the glutathione degradation protein and glutathione uptake protein was constructed. This strain improved the extracellular glutathione productivity by up to 3.2-fold compared to the parental strain. Second, the ATP-dependent permease Adp1 was identified as a novel glutathione export ABC protein (Gxa1) in S. cerevisiae based on the homology of the protein sequence with that of the known human glutathione export ABC protein (ABCG2). Overexpression of this GXA1 gene improved the extracellular glutathione production by up to 2.3-fold compared to the platform host strain. Finally, combinatorial overexpression of the GXA1 gene and the genes involved in glutathione synthesis in the platform host strain increased the extracellular glutathione production by up to 17.1-fold compared to the parental strain. Overall, the metabolic engineering of the glutathione synthesis, degradation, and transport increased the total (extracellular + intracellular) glutathione production. The extracellular glutathione fermentation method developed in this study has the potential to overcome the limitations of the present intracellular glutathione fermentation process in yeast.

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Acknowledgments

We are grateful to Dr. J. Ishii (Organization of Advanced Science and Technology, Kobe University) for providing us with the pGK plasmid series. This study was supported by the Special Coordination Funds for Promoting Science and Technology, Creation of Innovation Centers for Advanced Interdisciplinary Research Areas (Innovative Bioproduction Kobe), MEXT, Japan. K.Y. Hara was supported by a Grant-in-Aid for Young Scientists (B) (22760608).

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Authors and Affiliations

  1. Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, 657-8501, Japan

    Kentaro Kiriyama & Akihiko Kondo

  2. Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, 657-8501, Japan

    Kiyotaka Y. Hara

Authors
  1. Kentaro Kiriyama
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  2. Kiyotaka Y. Hara
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  3. Akihiko Kondo
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Corresponding author

Correspondence to Akihiko Kondo.

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Kentaro Kiriyama and Kiyotaka Y. Hara contributed equally to this work.

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Kiriyama, K., Hara, K.Y. & Kondo, A. Extracellular glutathione fermentation using engineered Saccharomyces cerevisiae expressing a novel glutathione exporter. Appl Microbiol Biotechnol 96, 1021–1027 (2012). https://doi.org/10.1007/s00253-012-4075-3

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  • DOI: https://doi.org/10.1007/s00253-012-4075-3

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