期刊
PROCESS BIOCHEMISTRY
卷 47, 期 5, 页码 853-857出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.procbio.2012.02.024
关键词
Module sub-division; CBM; Transplanting; Xylanase
资金
- Natural Science Foundation of China [30972123, 20971050]
Transplantation is useful for elucidating the functions of structural modules and for engineering enzyme properties. Unexpectedly, transplanting a hyper-thermophilic carbohydrate-binding module, CBM9_1-2, into the mesophilic Aspergillus niger GH11 xylanase (Xyn) slightly decreased the thermal inactivation half-life of Xyn. This effect was further investigated by dividing the CBM9_1-2 module into two smaller parts, C1 and C2, which were transplanted into Xyn to create the, chimeras Xyn-C1 and Xyn-C2. Both chimeras exhibited higher catalytic activities on xylan than native Xyn. Xyn-C2 exhibited higher binding affinities for both oat spelt and birch wood xylans, and its thermal inactivation half-life (69.3 min) was 4 or 5 times longer than that of Xyn (17.6 min), Xyn-C1 (13.4 min), and the original chimera containing CBM9_1-2 (13.8 min). In contrast, Xyn-C1 exhibited higher binding affinity for oat spelt xylan, but not for birch wood xylan. Through this rational engineering of the fungal xylanase, the C2 sub-module was shown to have a different thermostabilizing effect than the Cl sub-module. The different functions of the smaller parts of a large module can play pivotal roles in transplantation. (C) 2012 Elsevier Ltd. All rights reserved.
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