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Gene cloning, expression, and X-ray crystallographic analysis of a β-mannanase from Eisenia fetida
https://repo.qst.go.jp/records/49415
https://repo.qst.go.jp/records/49415c0893d9c-b17a-4c1b-8ba7-d2ad7e9d402f
| Item type | 学術雑誌論文 / Journal Article(1) | |||||
|---|---|---|---|---|---|---|
| 公開日 | 2019-01-09 | |||||
| タイトル | ||||||
| タイトル | Gene cloning, expression, and X-ray crystallographic analysis of a β-mannanase from Eisenia fetida | |||||
| 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 |
上田, 光宏
× 上田, 光宏× 平野, 優× 福原, 宏章× 中, 裕規× 中澤, 昌美× 阪本, 龍司× Ogata, Yoshiyuki× 玉田, 太郎× 平野 優× 玉田 太郎 |
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| 抄録 | ||||||
| 内容記述タイプ | Abstract | |||||
| 内容記述 | The Ef-Man gene was determined to consist of 1131 bp and encode a 377 amino acid protein. The amino acid sequence showed similarity with the endo-1,4-beta-mannanases of Daphnia pulex (62%), Cryptopygus antarcticus (64%), Crassostrea gigas (61%), Mytilus edulis (60%), and Aplisia kurodai (58%). The gene encoding mature Ef-Man was expressed in Pichia pastoris (GS115 strain). Based on SDS-PAGE analysis, the molecular mass of the purified recombinant Ef-Man (rEf-Man) was estimated to be 39.5 kDa. All catalytically important residues of endo-1,4-beta-mannanases in the glycoside hydrolase (GH) family 5 were conserved in Ef-Man. The optimal temperature for rEf-Man was identified as 60°C. HPLC and HPAEC analyses suggest that Ef-Man requires at least six subsites for efficient hydrolysis and is capable of performing transglycosylation reactions. The overall structure of rEf-Man is similar to those of GH5 family proteins, and tertiary structures around the active site are conserved among endo-1,4-beta-mannanase families. X-ray crystallographic analysis supports the hydrolysis and transglycosylation reaction mechanism determined by HPLC and HPAEC analyses. | |||||
| 書誌情報 |
Enzyme and Microbial Technology 巻 117, p. 15-22, 発行日 2018-05 |
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| ISSN | ||||||
| 収録物識別子タイプ | ISSN | |||||
| 収録物識別子 | 0141-0229 | |||||
| DOI | ||||||
| 識別子タイプ | DOI | |||||
| 関連識別子 | 10.1016/j.enzmictec.2018.05.014 | |||||
| 関連サイト | ||||||
| 識別子タイプ | URI | |||||
| 関連識別子 | https://www.sciencedirect.com/science/article/pii/S0141022918301984?via%3Dihub | |||||
| 関連名称 | https://www.sciencedirect.com/science/article/pii/S0141022918301984?via%3Dihub | |||||
