Unexpected regioselectivity of Humicola insolens Cel7B glycosynthase mutants

被引：11
作者：
Blanchard, Sophie
Armand, Sylvie
Couthino, Pedro
Patkar, Shamkant
Vind, Jesper
Samain, Eric
Driguez, Hugues
Cottaz, Sylvain
机构：
[1] CNRS, FR 2607, CERMAV, F-38041 Grenoble, France
[2] AFMB, UMR 6098, F-13288 Marseille, France
[3] NOVOZYMES, DK-2880 Bagsvaerd, Denmark
来源：
CARBOHYDRATE RESEARCH | 2007年 / 342卷 / 05期
关键词：
glycosynthase; cellulase; regioselectivity; enzymatic synthesis;
D O I：
10.1016/j.carres.2006.12.017
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：
Four Humicola insolens Ce17B glycoside hydrolase mutants have been evaluated for the coupling of lactosyl fluoride on O-allyl N-I-acetyl-2(II)-azido-beta-chitobioside. Double mutants Ce17B E197A H209A and Ce1713 E197A H209G preferentially catalyze the formation of a beta-(1 -> 4) linkage between the two disaccharides, while single mutant Cel7B E197A and triple mutant Cel7B E197A H209A A211T produce predominantly the beta-(1 -> 3)-linked tetrasaccharide. This result constitutes the first report of the modulation of the regioselectivity through site-directed mutagenesis for an endoglycosynthase. (c) 2007 Elsevier Ltd. All rights reserved.
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收藏
页码：710 / 716
页数：7
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