Engineering Escherichia coli for efficient glutathione production.

Autor: Mori H; Agri-Bio Research Center, KANEKA CORPORATION, 1-8, Miyamae-cho, Takasago-cho, Takasago, Hyogo, 676-8688, Japan., Matsui M; Agri-Bio Research Center, KANEKA CORPORATION, 1-8, Miyamae-cho, Takasago-cho, Takasago, Hyogo, 676-8688, Japan., Bamba T; Engineering Biology Research Center, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan., Hori Y; Engineering Biology Research Center, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan., Kitamura S; Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5, Yamadaoka, Suita, Osaka, 565-0871, Japan., Toya Y; Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5, Yamadaoka, Suita, Osaka, 565-0871, Japan., Hidese R; Engineering Biology Research Center, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan., Yasueda H; Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan; Research and Development Center for Precision Medicine, University of Tsukuba, 1-2 Kasuga, Tsukuba-shi, Ibaraki, 305-8550, Japan., Hasunuma T; Engineering Biology Research Center, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan; Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan; RIKEN Center for Sustainable Resource Science, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan., Shimizu H; Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5, Yamadaoka, Suita, Osaka, 565-0871, Japan., Taoka N; Agri-Bio Research Center, KANEKA CORPORATION, 1-8, Miyamae-cho, Takasago-cho, Takasago, Hyogo, 676-8688, Japan., Kobayashi S; Agri-Bio Research Center, KANEKA CORPORATION, 1-8, Miyamae-cho, Takasago-cho, Takasago, Hyogo, 676-8688, Japan. Electronic address: Shingo.Kobayashi@kaneka.co.jp.
Jazyk: angličtina
Zdroj: Metabolic engineering [Metab Eng] 2024 Jul; Vol. 84, pp. 180-190. Date of Electronic Publication: 2024 Jul 03.
DOI: 10.1016/j.ymben.2024.07.001
Abstrakt: Glutathione is a tripeptide of excellent value in the pharmaceutical, food, and cosmetic industries that is currently produced during yeast fermentation. In this case, glutathione accumulates intracellularly, which hinders high production. Here, we engineered Escherichia coli for the efficient production of glutathione. A total of 4.3 g/L glutathione was produced by overexpressing gshA and gshB, which encode cysteine glutamate ligase and glutathione synthetase, respectively, and most of the glutathione was excreted into the culture medium. Further improvements were achieved by inhibiting degradation (Δggt and ΔpepT); deleting gor (Δgor), which encodes glutathione oxide reductase; attenuating glutathione uptake (ΔyliABCD); and enhancing cysteine production (P ompF -cysE). The engineered strain KG06 produced 19.6 g/L glutathione after 48 h of fed-batch fermentation with continuous addition of ammonium sulfate as the sulfur source. We also found that continuous feeding of glycine had a crucial role for effective glutathione production. The results of metabolic flux and metabolomic analyses suggested that the conversion of O-acetylserine to cysteine is the rate-limiting step in glutathione production by KG06. The use of sodium thiosulfate largely overcame this limitation, increasing the glutathione titer to 22.0 g/L, which is, to our knowledge, the highest titer reported to date in the literature. This study is the first report of glutathione fermentation without adding cysteine in E. coli. Our findings provide a great potential of E. coli fermentation process for the industrial production of glutathione.
(Copyright © 2024 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: