論文 - 詳細
| RRC ID | 34514 |
|---|---|
| 著者 | Koyama Y, Hidaka M, Nishimoto M, Kitaoka M. |
| タイトル | Directed evolution to enhance thermostability of galacto-N-biose/lacto-N-biose I phosphorylase. |
| ジャーナル | Protein Eng Des Sel |
| Abstract |
Galacto-N-biose/lacto-N-biose I phosphorylase (GLNBP) is the key enzyme in the enzymatic production of lacto-N-biose I. For the purpose of industrial use, we improved the thermostability of GLNBP by evolutionary engineering in which five substitutions in the amino acid sequence were selected from a random mutagenesis GLNBP library constructed using error-prone polymerase chain reaction. Among them, C236Y and D576V mutants showed considerably improved thermostability. Structural analysis of C236Y revealed that the hydroxyl group of Tyr236 forms a hydrogen bond with the carboxyl group of E319. The C236Y and D576V mutations together contributed to the thermostability. The C236Y/D576V mutant exhibited 20°C higher thermostability than the wild type. |
| 巻・号 | 26(11) |
| ページ | 755-61 |
| 公開日 | 2013-11-1 |
| DOI | 10.1093/protein/gzt049 |
| PII | gzt049 |
| PMID | 24065834 |
| MeSH | Amino Acid Sequence Directed Molecular Evolution / methods* Enzyme Stability Hot Temperature Molecular Sequence Data Phosphorylases / chemistry* Phosphorylases / genetics* Phosphorylases / metabolism Sequence Alignment |
| IF | 1.774 |
| 引用数 | 10 |
| WOS 分野 | BIOTECHNOLOGY & APPLIED MICROBIOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY |
| リソース情報 | |
| 一般微生物 | JCM 1217 |