Cloning, expression and structural stability of a cold-adapted β-galactosidase from Rahnella sp R3

被引：38

作者：

Fan, Yuting ^{[1
,2
]}

Hua, Xiao ^{[1
]}

Zhang, Yuzhu ^{[3
]}

Feng, Yinghui ^{[2
]}

Shen, Qiuyun ^{[2
]}

Dong, Juan ^{[1
]}

Zhao, Wei ^{[2
]}

Zhang, Wenbin ^{[2
]}

Jin, Zhengyu ^{[1
]}

Yang, Ruijin ^{[1
,2
]}

机构：

[1] Jiangnan Univ, State Key Lab Food Sci & Technol, Wuxi 214122, Peoples R China

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

[3] ARS, USDA, Pacific West Area, Western Reg Res Ctr, Albany, CA 94710 USA

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2015年 / 115卷

关键词：

beta-Galactosidase; Cold-adapted enzyme; Enzyme structure stability; Lactose hydrolysis; BIOCHEMICAL-CHARACTERIZATION; GENOME SEQUENCE; PURIFICATION; STRAIN; AQUATILIS;

D O I：

10.1016/j.pep.2015.07.001

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A novel gene was isolated for the first time from a psychrophilic gram-negative bacterium Rahnella sp. R3. The gene encoded a cold-adapted beta-galactosidase (R-beta-Gal). Recombinant R-beta-Gal was expressed in Escherichia coli BL21 (DE3), purified and characterized. R-beta-gal belongs to the glycosyl hydrolase family 42. Circular dichroism spectrometry of the structural stability of R-beta-Gal with respect to temperature indicated that the secondary structures of the enzyme were stable to 45 degrees C. In solution, the enzyme was a homo-trimer and was active at temperatures as low as 4 degrees C. The enzyme did not require the presence of metal ions to be active, but Mg2+, Mn2+, and Ca2+ enhanced its activity slightly, whereas Fe3+, Zn2+ and Al3+ appeared to inactive it. The purified enzyme displayed K-m values of 6.5 mM for ONPG and 2.2 mM for lactose at 4 degrees C. These values were lower than the corresponding K(m)s reported for other cold-adapted beta-Gals. (C) 2015 Elsevier Inc. All rights reserved.

引用

页码：158 / 164

页数：7

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