論文 - 詳細
| RRC ID | 71014 |
|---|---|
| 著者 | Xia W, Bai Y, Shi P. |
| タイトル | Improving the Substrate Affinity and Catalytic Efficiency of β-Glucosidase Bgl3A from Talaromyces leycettanus JCM12802 by Rational Design. |
| ジャーナル | Biomolecules |
| Abstract |
Improving the substrate affinity and catalytic efficiency of β-glucosidase is necessary for better performance in the enzymatic saccharification of cellulosic biomass because of its ability to prevent cellobiose inhibition on cellulases. Bgl3A from Talaromyces leycettanus JCM12802, identified in our previous work, was considered a suitable candidate enzyme for efficient cellulose saccharification with higher catalytic efficiency on the natural substrate cellobiose compared with other β-glucosidase but showed insufficient substrate affinity. In this work, hydrophobic stacking interaction and hydrogen-bonding networks in the active center of Bgl3A were analyzed and rationally designed to strengthen substrate binding. Three vital residues, Met36, Phe66, and Glu168, which were supposed to influence substrate binding by stabilizing adjacent binding site, were chosen for mutagenesis. The results indicated that strengthening the hydrophobic interaction between stacking aromatic residue and the substrate, and stabilizing the hydrogen-bonding networks in the binding pocket could contribute to the stabilized substrate combination. Four dominant mutants, M36E, M36N, F66Y, and E168Q with significantly lower Km values and 1.4-2.3-fold catalytic efficiencies, were obtained. These findings may provide a valuable reference for the design of other β-glucosidases and even glycoside hydrolases. |
| 巻・号 | 11(12) |
| 公開日 | 2021-12-15 |
| DOI | 10.3390/biom11121882 |
| PII | biom11121882 |
| PMID | 34944526 |
| PMC | PMC8699594 |
| MeSH | Binding Sites Biocatalysis Cellobiose / metabolism* Fungal Proteins / chemistry Fungal Proteins / genetics Fungal Proteins / metabolism Hydrogen Bonding Hydrogen-Ion Concentration Models, Molecular Molecular Dynamics Simulation Mutation* Protein Conformation Substrate Specificity Talaromyces / chemistry Talaromyces / enzymology* beta-Glucosidase / chemistry* beta-Glucosidase / genetics beta-Glucosidase / metabolism* |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 1 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| 一般微生物 | JCM12802 |