A novel cold-adapted and glucose-tolerant GH1 β-glucosidase from Exiguobacterium antarcticum B7

被引：53

作者：

Crespim, Elaine ^{[1
]}

Zanphorlin, Leticia M. ^{[1
]}

de Souza, Flavio H. M. ^{[2
]}

Diogo, Jose A. ^{[1
]}

Gazolla, Alex C. ^{[1
]}

Machado, Carla B. ^{[1
]}

Figueiredo, Fernanda ^{[1
]}

Sousa, Amanda S. ^{[1
]}

Nobrega, Felipe ^{[3
]}

Pellizari, Vivian H. ^{[3
]}

Murakami, Mario T. ^{[2
]}

Ruller, Roberto ^{[1
]}

机构：

[1] CNPEM, Lab Nacl Ciencia & Tecnol Bioetanol CTBE, BR-13083970 Campinas, SP, Brazil

[2] CNPEM, Lab Nacl Biociencias LNBio, BR-13083970 Campinas, SP, Brazil

[3] Univ Sao Paulo, Inst Oceanog, Sao Paulo, SP, Brazil

来源：

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES | 2016年 / 82卷

基金：

巴西圣保罗研究基金会;

关键词：

beta-Glucosidase; Cold adaptation; Glucose tolerance; Exiguobacterium antarcticum B7; STABILITY RELATIONSHIPS; MICROBIAL METAGENOME; MOLECULAR-CLONING; PURIFICATION; CELLULASE; FERMENTATION; HYDROLYSIS; EXPRESSION; HYDROLASES; SEQUENCE;

D O I：

10.1016/j.ijbiomac.2015.09.018

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A novel GH1 beta-glucosidase (EaBgl1 A) from a bacterium isolated from Antarctica soil samples was recombinantly overexpressed in Escherichia coil cells and characterized. The enzyme showed unusual pH dependence with maximum activity at neutral pH and retention of high catalytic activity in the pH range 6 to 9, indicating a catalytic machinery compatible with alkaline conditions. EaBgl1 A is also a coldadapted enzyme, exhibiting activity in the temperature range from 10 to 40 degrees C with optimal activity at 30 degrees C, which allows its application in industrial processes using low temperatures. Kinetic characterization revealed an enzymatic turnover (K-cat) of 6.92s(-1) (cellobiose) and 32.98 s(-1) (pNPG) and a high tolerance for product inhibition, which is an extremely desirable feature for biotechnological purposes. Interestingly, the enzyme was stimulated by up to 200mM glucose, whereas the commercial cocktails tested were found fully inhibited at this concentration. These properties indicate EaBgl1A as a promising biocatalyst for biotechnological applications where low temperatures are required. (c) 2015 Elsevier B.V. All rights reserved.

引用

页码：375 / 380

页数：6

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