caa a22 4500 605500908 CHVBK 20210128100558.0 cr unu---uuuuu 210128e20151101xx s 000 0 eng 10.1007/s00253-015-6847-z doi (NATIONALLICENCE)springer-10.1007/s00253-015-6847-z Improvement of oxidized glutathione fermentation by thiol redox metabolism engineering in Saccharomyces cerevisiae [Elektronische Daten] [Kiyotaka Hara, Naoko Aoki, Jyumpei Kobayashi, Kentaro Kiriyama, Keiji Nishida, Michihiro Araki, Akihiko Kondo] 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. Springer-Verlag Berlin Heidelberg, 2015 Oxidized glutathione nationallicence Thiol oxidase nationallicence Yeast nationallicence Saccharomyces cerevisiae nationallicence Metabolic engineering nationallicence Cell factory nationallicence Hara Kiyotaka Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai-cho, 657-8501, Kobe, Nada-ku, Japan aut Aoki Naoko Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai-cho, 657-8501, Kobe, Nada-ku, Japan aut Kobayashi Jyumpei Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai-cho, 657-8501, Kobe, Nada-ku, Japan aut Kiriyama Kentaro Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodaicho, 657-8501, Kobe, Nada-ku, Japan aut Nishida Keiji Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai-cho, 657-8501, Kobe, Nada-ku, Japan aut Araki Michihiro Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai-cho, 657-8501, Kobe, Nada-ku, Japan aut Kondo Akihiko Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodaicho, 657-8501, Kobe, Nada-ku, Japan aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/22(2015-11-01), 9771-9778 0175-7598 99:22<9771 2015 99 253 https://doi.org/10.1007/s00253-015-6847-z text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6847-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Hara Kiyotaka Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai-cho, 657-8501, Kobe, Nada-ku, Japan aut NATIONALLICENCE 700 1- Aoki Naoko Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai-cho, 657-8501, Kobe, Nada-ku, Japan aut NATIONALLICENCE 700 1- Kobayashi Jyumpei Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai-cho, 657-8501, Kobe, Nada-ku, Japan aut NATIONALLICENCE 700 1- Kiriyama Kentaro Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodaicho, 657-8501, Kobe, Nada-ku, Japan aut NATIONALLICENCE 700 1- Nishida Keiji Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai-cho, 657-8501, Kobe, Nada-ku, Japan aut NATIONALLICENCE 700 1- Araki Michihiro Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai-cho, 657-8501, Kobe, Nada-ku, Japan aut NATIONALLICENCE 700 1- Kondo Akihiko Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodaicho, 657-8501, Kobe, Nada-ku, Japan aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/22(2015-11-01), 9771-9778 0175-7598 99:22<9771 2015 99 253