Improved thermostability of a metagenomic glucose-tolerant β-glycosidase based on its X-ray crystal structure

被引：13

作者：

Matsuzawa, Tomohiko ^{[1
]}

Watanabe, Masahiro ^{[2
,3
]}

Yaoi, Katsuro ^{[1
]}

机构：

[1] Natl Inst Adv Ind Sci & Technol, Bioprod Res Inst, Tsukuba Cent 6,1-1-1 Higashi, Tsukuba, Ibaraki 3058566, Japan

[2] Natl Inst Adv Ind Sci & Technol, Biomass Refinery Res Ctr, 3-11-32 Kagamiyama, Higashihiroshima, Hiroshima 7390046, Japan

[3] Natl Inst Adv Ind Sci & Technol, Res Inst Sustainable Chem, 3-11-32 Kagamiyama, Higashihiroshima, Hiroshima 7390046, Japan

来源：

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY | 2017年 / 101卷 / 23-24期

关键词：

beta-glycosidase; Metagenome; Biomass; Thermostabilization; TRICHODERMA-REESEI; PICHIA-PASTORIS; AMINO-ACID; EXPRESSION; IDENTIFICATION; ADSORPTION; STABILITY; CLONING; GENE; CELLOBIOHYDROLASES;

D O I：

10.1007/s00253-017-8525-9

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

MeBglD2, a metagenomic beta-glycosidase, is stimulated by various saccharides, including d-glucose, d-xylose, and maltose, and it promotes the enzymatic saccharification of plant biomass. To improve the thermostability of MeBglD2, its X-ray crystal structure was analyzed, and the amino acid residues responsible for its thermostability were identified using the structural information. Mutations in His8, Asn59, and Gly295 improved the thermostability of MeBglD2, and the combination of these mutations resulted in the highest thermostability. Compared with wild-type MeBglD2, thermostable MeBglD2 mutants promoted plant biomass saccharification using Trichoderma reesei cellulase. In addition to thermostability, the thermostable mutants exhibited higher tolerance to ethanol, dimethyl sulfoxide, and copper ions, indicating that the MeBglD2 mutants generated in this study were improved in their tolerance to not only high temperature but also to organic solvents and metal ions.

引用

页码：8353 / 8363

页数：11

共 51 条

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