Swapping the chitin-binding domain in Bacillus chitinases improves the substrate binding affinity and conformational stability

被引：11

作者：

Neeraja, Chilukoti ^{[1
,3
]}

Subramanyam, Rajagopal ^{[2
]}

Moerschbacher, Bruno M. ^{[3
]}

Podile, Appa Rao ^{[1
]}

机构：

[1] Univ Hyderabad, Dept Plant Sci, Hyderabad 500046, Andhra Pradesh, India

[2] Univ Hyderabad, Dept Biochem, Hyderabad 500046, Andhra Pradesh, India

[3] Univ Munster, Dept Plant Biochem & Biotechnol, D-48143 Munster, Germany

来源：

MOLECULAR BIOSYSTEMS | 2010年 / 6卷 / 08期

关键词：

THURINGIENSIS SUBSP KURSTAKI; SECONDARY STRUCTURE; CIRCULANS WL-12; ESCHERICHIA-COLI; LICHENIFORMIS; GENE; A1; PURIFICATION; SPECTROSCOPY; EXPRESSION;

D O I：

10.1039/b923048c

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：

Chitinase from Bacillus thuringiensis and Bacillus licheniformis consisting of an N-terminal catalytic domain (GH18) and a C-terminal chitin-binding domain (ChBD), were cloned and characterised. In order to study the importance of individual domains, chimeric chitinases (BtGH-BliChBD and BliGH-BtChBD) were constructed using domain swapping as a strategy to exchange the CBD of BtGH-ChBD with that of BliGH-ChBD and vice versa. Both chimeric chitinases showed increased affinity to colloidal chitin. BtGH-BliChBD was different from the three other chitinases studied concerning optimum temperature and pH. Additionally, BtGH-BliChBD and BliGH-BtChBD showed significant improvement in functional stability, conformational stability, and binding ability towards insoluble chitinous substrates compared to those of the native chitinases.

引用

页码：1492 / 1502

页数：11

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