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https://hdl.handle.net/20.500.14094/90004184
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2024-04-27
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90004184 (fulltext)
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メタデータID
90004184
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open access
出版タイプ
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タイトル
Enzymatic improvement of mitochondrial thiol oxidase Erv1 for oxidized glutathione fermentation by Saccharomyces cerevisiae
著者
Kobayashi, Jyumpei ; Sasaki, Daisuke ; Hara, Kiyotaka Y. ; Hasunuma, Tomohisa ; Kondo, Akihiko
著者ID
A0224
研究者ID
1000090759987
著者名
Kobayashi, Jyumpei
小林, 淳平
コバヤシ, ジュンペイ
所属機関名
科学技術イノベーション研究科
著者ID
A0276
研究者ID
1000000650615
著者名
Sasaki, Daisuke
佐々木, 大介
ササキ, ダイスケ
所属機関名
科学技術イノベーション研究科
著者名
Hara, Kiyotaka Y.
著者ID
A0960
研究者ID
1000020529606
KUID
https://kuid-rm-web.ofc.kobe-u.ac.jp/search/detail?systemId=73b63639d47d0a4b520e17560c007669
著者名
Hasunuma, Tomohisa
蓮沼, 誠久
ハスヌマ, トモヒサ
所属機関名
先端バイオ工学研究センター
著者ID
A1715
研究者ID
1000040205547
KUID
https://kuid-rm-web.ofc.kobe-u.ac.jp/search/detail?systemId=a324eb4a1b052e53520e17560c007669
著者名
Kondo, Akihiko
近藤, 昭彦
コンドウ, アキヒコ
所属機関名
科学技術イノベーション研究科
収録物名
Microbial Cell Factories
巻(号)
16
ページ
44-44
出版者
BioMed Central
刊行日
2017-03-15
公開日
2017-09-14
抄録
Background: Oxidized glutathione (GSSG) is the preferred form for industrial mass production of glutathione due to its high stability compared with reduced glutathione (GSH). In our previous study, over-expression of the mitochondrial thiol oxidase ERV1 gene was the most effective for high GSSG production in Saccharomyces cerevisiae cells among three types of different thiol oxidase genes. Results: We improved Erv1 enzyme activity for oxidation of GSH and revealed that S32 and N34 residues are critical for the oxidation. Five engineered Erv1 variant proteins containing S32 and/or N34 replacements exhibited 1.7- to 2.4-fold higher in vitro GSH oxidation activity than that of parental Erv1, whereas the oxidation activities of these variants for γ-glutamylcysteine were comparable. According to three-dimensional structures of Erv1 and protein stability assays, S32 and N34 residues interact with nearby residues through hydrogen bonding and greatly contribute to protein stability. These results suggest that increased flexibility by amino acid replacements around the active center decrease inhibitory effects on GSH oxidation. Over-expressions of mutant genes coding these Erv1 variants also increased GSSG and consequently total glutathione production in S. cerevisiae cells. Over-expression of the ERV1(S32A) gene was the most effective for GSSG production in S. cerevisiae cells among the parent and other mutant genes, and it increased GSSG production about 1.5-fold compared to that of the parental ERV1 gene. Conclusions: This is the first study demonstrating the pivotal effects of S32 and N34 residues to high GSH oxidation activity of Erv1. Furthermore, in vivo validity of Erv1 variants containing these S32 and N34 replacements were also demonstrated. This study indicates potentials of Erv1 for high GSSG production.
カテゴリ
科学技術イノベーション研究科
先端バイオ工学研究センター
学術雑誌論文
権利
© The Author(s) 2017.
This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
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資源タイプ
journal article
言語
English (英語)
eISSN
1475-2859
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DOI
https://doi.org/10.1186/s12934-017-0658-0
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