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Mutation of Trp137 to glutamate completely removes transglycosyl activity associated with the Aspergillus fumigatus AfChiB1
Lue, Yang ; Yang, Haitao ; Hu, Hongyan ; Wang, Ying ; Rao, Zihe ; Jin, Cheng
2010-10-12 ; 2010-10-12
关键词Chitinase Transglycosylation Aspergillus fumigatus Mutation SUBSTRATE-ASSISTED CATALYSIS HUMAN MACROPHAGE CHITINASE NOCARDIA-ORIENTALIS GLYCOSYL HYDROLASE CRYSTAL-STRUCTURES ALLOSAMIDIN LYSOZYME COMPLEX CHITOTRIOSIDASE HYDROLYSIS Biochemistry & Molecular Biology
中文摘要Family 18 chitinases hydrolyze chitin through a substrate-assisted catalytic mechanism and are to a variable extent able to catalyze transglycosylation reactions. Previously Aspergillus fumigatus AfChiB1 was found to be able to catalyze transglycosylation reactions. Structural analysis reveals that AfChiB1 consists of an eight-stranded beta/alpha-barrel. Like other members of the family 18 hydrolases, AfChiB1 has conserved substrate binding site and catalytic acid, while a suitable nucleophile is missing. In this study, Trp137, Asp246, and Met243, which are close to the glycosidic cleavage site, were mutated to glutamate individually. As a result, the W137E remained its hydrolytic activity and was completely devoid of transglycosyl activity, while the D246E reduced its chitinolytic activity and increased its transglycosyl activity. And the M243E showed a remarkable reduction of chitinolytic activity and complete loss of transglycosyl activity. These results suggested that the transglycosyl reaction catalyzed by the AfChiB1 is due to lacking of nucleophile.
语种英语 ; 英语
出版者SPRINGER ; DORDRECHT ; VAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDS
内容类型期刊论文
源URL[http://hdl.handle.net/123456789/81835]  
专题清华大学
推荐引用方式
GB/T 7714
Lue, Yang,Yang, Haitao,Hu, Hongyan,et al. Mutation of Trp137 to glutamate completely removes transglycosyl activity associated with the Aspergillus fumigatus AfChiB1[J],2010, 2010.
APA Lue, Yang,Yang, Haitao,Hu, Hongyan,Wang, Ying,Rao, Zihe,&Jin, Cheng.(2010).Mutation of Trp137 to glutamate completely removes transglycosyl activity associated with the Aspergillus fumigatus AfChiB1..
MLA Lue, Yang,et al."Mutation of Trp137 to glutamate completely removes transglycosyl activity associated with the Aspergillus fumigatus AfChiB1".(2010).
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