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Expression and characterization of l-arabinose isomerase from Geobacillus stearothermophilus for improved activity under acidic condition
https://repo.qst.go.jp/records/80893
https://repo.qst.go.jp/records/808933293bd53-4850-4600-81da-4e8ebeb1f5b4
| Item type | 学術雑誌論文 / Journal Article(1) | |||||
|---|---|---|---|---|---|---|
| 公開日 | 2020-10-28 | |||||
| タイトル | ||||||
| タイトル | Expression and characterization of l-arabinose isomerase from Geobacillus stearothermophilus for improved activity under acidic condition | |||||
| 言語 | ||||||
| 言語 | eng | |||||
| 資源タイプ | ||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
| 資源タイプ | journal article | |||||
| アクセス権 | ||||||
| アクセス権 | metadata only access | |||||
| アクセス権URI | http://purl.org/coar/access_right/c_14cb | |||||
| 著者 |
Amreta Laksmi, Fina
× Amreta Laksmi, Fina× Arai, Shigeki× Arakawa, Tsutomu× Tsurumaru, Hirohito× Nakamura, Yoshitaka× Sakson, Budi× Tokunaga, Masao× Ishibashi, Matsujiro× Shigeki, Arai |
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| 抄録 | ||||||
| 内容記述タイプ | Abstract | |||||
| 内容記述 | A low-calorie sugar-substituting sweetener, d-tagatose, can be produced by l-arabinose isomerase (l-AI) from the substrate d-galactose. However, this process suffers from a Maillard reaction when performed at alkaline pH and high temperature. For industrial applications, therefore, a reaction under slightly acidic conditions is desirable to minimize the Maillard reaction. Previously, we obtained a mutant of l-AI, H18T, from Geobacillus stearothermophilus with greater substrate specificity. Although H18T possessed excellent thermostability, its activity under acidic conditions was not optimal. Here, we successfully obtained a potential variant of the H18T protein, H18T-Y234C, which achieved improved activity at pH 6.0, based on random mutagenesis using error-prone PCR around the binding pocket area of H18T. This double H18T-Y234C mutant possessed 1.8-fold and 3-fold higher activity at pH 6.0 than the parent H18T and the wild type, thereby broadening the optimal pH range to 6.0-8.0. Mutation from Tyr to Cys at residue 234 had little effect on the secondary structure of L-AI. Furthermore, the formation of disulfide bonds was not detected. Thus, the improvement of activity at pH 6.0 is probably caused by the change in the binding pocket area involving residue 234. This study offers insight into the importance of residue 234 in improving the activity under acidic conditions. | |||||
| 書誌情報 |
Protein Expression and Purification 巻 175, p. 105692, 発行日 2020-07 |
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| 出版者 | ||||||
| 出版者 | Elsevier | |||||
| ISSN | ||||||
| 収録物識別子タイプ | ISSN | |||||
| 収録物識別子 | 1046-5928 | |||||
| PubMed番号 | ||||||
| 識別子タイプ | PMID | |||||
| 関連識別子 | 32681957 | |||||
| DOI | ||||||
| 識別子タイプ | DOI | |||||
| 関連識別子 | 10.1016/j.pep.2020.105692 | |||||
| 関連サイト | ||||||
| 識別子タイプ | URI | |||||
| 関連識別子 | https://www.sciencedirect.com/science/article/abs/pii/S1046592820302837?via%3Dihub | |||||
