Fusion of cellulose binding domain to the catalytic domain improves the activity and conformational stability of chitinase in Bacillus licheniformis DSM13
Fusion of cellulose binding domain to the catalytic domain improves the activity and conformational stability of chitinase in Bacillus licheniformis DSM13
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DOI：
10.1016/j.biortech.2009.12.118
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发表时间：
2010-05-01
影响因子：
11.4
通讯作者：
Podile, Appa Rao
中科院分区：
文献类型：
--
作者：
Neeraja, Chilukoti;Moerschbacher, Bruno;Podile, Appa Rao
地衣芽孢杆菌（Bacillus licheniformis）DSM 13的几丁质酶由N端催化结构域（GH）和C端几丁质结合结构域（ChBD）组成。利用聚合酶链反应（PCR）技术，构建了缺失突变体BliGH和杂交几丁质酶BliGH-CeBD，研究了底物结合结构域的作用。这两种重组几丁质酶保留了它们与乙二醇几丁质（GC）结合的能力。BliGH对胶体几丁质（CC）比BliGH-CeBD更有效，如从增加的V-max和k（cat）值所证明的。与缺失突变体BliGH相比，融合基因BliGH-CeBD对胶体几丁质的亲和性、活性和构象稳定性均有所提高。(c)2009爱思唯尔有限公司版权所有。
Chitinase from Bacillus licheniformis DSM13 consists of an N-terminal catalytic domain (GH) and a C-terminal chitin binding domain (ChBD). A deletion mutant BliGH and a hybrid chitinase BliGH-CeBD were developed using polymerase chain reaction (PCR) to study the role of substrate-binding domain. Both recombinant chitinases retained their ability to bind to glycol-chitin (GC). BliGH was more effective on colloidal chitin (CC) than BliGH-CeBD as evident from the increased V-max and k(cat) values. The fusion of CeBD improved the affinity to colloidal chitin, activity and conformational stability in BliGH-CeBD when compared with deletion mutant BliGH. (c) 2009 Elsevier Ltd. All rights reserved.