Expression and Biochemical Characterization of a Thermostable Branching Enzyme from Geobacillus thermoglucosidans

被引：19

作者：

Ban, Xiaofeng ^{[1
,2
]}

Li, Caiming ^{[1
,2
]}

Gu, Zhengbiao ^{[1
,2
,3
]}

Bao, Chunhui ^{[1
,2
]}

Qiu, Yijing ^{[1
,2
]}

Hong, Yan ^{[1
,2
,3
]}

Cheng, Li ^{[1
,2
]}

Li, Zhaofeng ^{[1
,2
,3
]}

机构：

[1] Jiangnan Univ, State Key Lab Food Sci & Technol, Wuxi, Jiangsu, Peoples R China

[2] Jiangnan Univ, Sch Food Sci & Technol, Wuxi 214122, Jiangsu, Peoples R China

[3] Jiangnan Univ, Collaborat Innovat Ctr Food Safety & Qual Control, Wuxi, Jiangsu, Peoples R China

来源：

JOURNAL OF MOLECULAR MICROBIOLOGY AND BIOTECHNOLOGY | 2016年 / 26卷 / 05期

基金：

美国国家科学基金会; 国家教育部博士点专项基金资助; 中国国家自然科学基金;

关键词：

Branching enzyme; Geobacillus thermoglucosidans; Thermostability; alpha-1,6 branch points; Starch; BACILLUS-STEAROTHERMOPHILUS; CYCLIZATION REACTION; NEUROSPORA-CRASSA; MOLECULAR-CLONING; ESCHERICHIA-COLI; GLYCOGEN; STARCH; GENE; AMYLOSE; MAIZE;

D O I：

10.1159/000446582

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：

The branching enzyme (EC 2.4.1.18) catalyzes the formation of alpha-1,6 branch points in starch. In this study, the Geobacillus thermoglucosidans gene-encoding branching enzyme was expressed in Escherichia coli BL21 (DE3) and the protein was isolated and characterized. G. thermoglucosidans branching enzyme is a thermostable enzyme with an optimal reaction temperature of nearly 60 degrees C and a half-life at 65 degrees C of approximately 1.1 h. The activity of the recombinant enzyme is optimal at pH 7.5, with broad stability between pH 5.5 and 9.0. Its thermostability, relatively broad pH stability and optimal temperature near the temperature at which starch begins to gelatinize may make it easy to use in industrial production. Furthermore, the enzyme is activated by Mg2+, Ba2+, K+ and Na+ in a concentration-dependent manner and dramatically inhibited by Ni2+ and Co2+. Its substrate dependence, using amylopectin as the substrate, could be adequately fitted using the Michaelis-Menten equation, yielding a K-m of 0.99 mg/ml. High-performance anion exchange chromatography results showed that the chain length distribution of branching enzyme-treated waxy corn starch is indistinguishable from that of the branching enzyme-treated common corn starch. This enzyme may therefore be a promising tool for the enzymatic modification of starch. (C) 2016 S. Karger AG, Basel

引用

页码：303 / 311

页数：9

共 52 条

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