α-glucosidase mutant catalyzes "α-glycosynthase"-type reaction

被引：76

作者：

Okuyama, M ^{[1
]}

Mori, H ^{[1
]}

Watanabe, K ^{[1
]}

Kimura, A ^{[1
]}

Chiba, S ^{[1
]}

机构：

[1] Hokkaido Univ, Grad Sch Agr, Div Appl Biosci, Sapporo, Hokkaido 0608589, Japan

来源：

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY | 2002年 / 66卷 / 04期

关键词：

alpha-glucosidase; catalytic nucleophile mutant; glycosynthase; oligosaccharide production;

D O I：

10.1271/bbb.66.928

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：

Replacement of the catalytic nucleophile Asp481 by glycine in Schizosaccharomyces pombe alpha-glucosidase eliminated the hydrolytic activity. The mutant enzyme (D481G) was found to catalyze the formation of an alpha-glucosidic linkage from beta-glucosyl fluoride and 4-nitrophenyl (PNP) alpha-glucoside to produce two kinds of PNP alpha-diglucosides, alpha-isomaltoside and alpha-maltoside. The two products were not hydrolyzed by D481G, giving 41 and 29% yields of PNP alpha-isomaltoside and alpha-maltoside, respectively. PNP monoglycosides, such as alpha-xyloside, alpha-mannoside, or beta-glucoside, acted as the substrate, but PNP alpha-galactoside and maltose could not. No detectable product was observed in the combination of alpha-glucosyl fluoride and PNP alpha-glucoside. This study is the first report on an "alpha-glycosynthase"-type reaction to form an alpha-glycosidic linkage.

引用

页码：928 / 933

页数：6

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