Applied Microbiology and Biotechnology

, Volume 99, Issue 22, pp 9771–9778 | Cite as

Improvement of oxidized glutathione fermentation by thiol redox metabolism engineering in Saccharomyces cerevisiae

  • Kiyotaka Y. Hara
  • Naoko Aoki
  • Jyumpei Kobayashi
  • Kentaro Kiriyama
  • Keiji Nishida
  • Michihiro Araki
  • Akihiko Kondo
Applied microbial and cell physiology

Abstract

Glutathione is a valuable tripeptide widely used in the pharmaceutical, food, and cosmetic industries. In industrial fermentation, glutathione is currently produced primarily using the yeast Saccharomyces cerevisiae. Intracellular glutathione exists in two forms; the majority is present as reduced glutathione (GSH) and a small amount is present as oxidized glutathione (GSSG). However, GSSG is more stable than GSH and is a more attractive form for the storage of glutathione extracted from yeast cells after fermentation. In this study, intracellular GSSG content was improved by engineering thiol oxidization metabolism in yeast. An engineered strain producing high amounts of glutathione from over-expression of glutathione synthases and lacking glutathione reductase was used as a platform strain. Additional over-expression of thiol oxidase (1.8.3.2) genes ERV1 or ERO1 increased the GSSG content by 2.9-fold and 2.0-fold, respectively, compared with the platform strain, without decreasing cell growth. However, over-expression of thiol oxidase gene ERV2 showed almost no effect on the GSSG content. Interestingly, ERO1 over-expression did not decrease the GSH content, raising the total glutathione content of the cell, but ERV1 over-expression decreased the GSH content, balancing the increase in the GSSG content. Furthermore, the increase in the GSSG content due to ERO1 over-expression was enhanced by additional over-expression of the gene encoding Pdi1, whose reduced form activates Ero1 in the endoplasmic reticulum. These results indicate that engineering the thiol redox metabolism of S. cerevisiae improves GSSG and is critical to increasing the total productivity and stability of glutathione.

Keywords

Oxidized glutathione Thiol oxidase Yeast Saccharomyces cerevisiae Metabolic engineering Cell factory 

Notes

Acknowledgments

We are grateful to Dr. J. Ishii (Organization of Advanced Science and Technology, Kobe University) for providing us with the pGK plasmid series. We also thank Dr. M. Mochizuki for technical assistance.

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2015_6847_MOESM1_ESM.pdf (134 kb)
ESM 1(PDF 134 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Kiyotaka Y. Hara
    • 1
    • 3
  • Naoko Aoki
    • 1
  • Jyumpei Kobayashi
    • 1
  • Kentaro Kiriyama
    • 2
  • Keiji Nishida
    • 1
  • Michihiro Araki
    • 1
  • Akihiko Kondo
    • 2
  1. 1.Organization of Advanced Science and TechnologyKobe UniversityKobeJapan
  2. 2.Department of Chemical Science and Engineering, Graduate School of EngineeringKobe UniversityKobeJapan
  3. 3.Graduate School of Nutritional and Environmental SciencesUniversity of ShizuokaShizuokaJapan

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