文章基本信息

标题：Conversion of a Rhizopus chinensis lipase into an esterase by lid swapping
本地全文：下载
作者：Xiao-Wei Yu ; Shan-Shan Zhu ; Rong Xiao 等
期刊名称：JLR Papers In Press
印刷版ISSN：0022-2275
电子版ISSN：1539-7262
出版年度：2014
卷号：55
期号：6
页码：1044-1051
DOI：10.1194/jlr.M043950
语种：English
出版社：American Society for Biochemistry and Molecular Biology
摘要：In an effort to explore the feasibility of converting a lipase into an esterase by modifying the lid region, we designed and characterized two novel Rhizopus chinensis lipase variants by lid swapping. The substrate specificity of an R. chinensis lipase was successfully modified toward water-soluble substrates, that is, turned into an esterase, by replacing the hydrophobic lid with a hydrophilic lid from ferulic acid esterase from Aspergillus niger . Meanwhile, as a comparison, the lid of R. chinensis lipase was replaced by a hydrophobic lid from Rhizomucor miehei lipase, which did not alter its substrate specificity but led to a 5.4-fold higher catalytic efficiency ( k * _cat / K * _m ) toward p -nitrophenyl laurate. Based on the analysis of structure-function relationships, it suggests that the amphipathic nature of the lid is very important for the substrate specificity. This study provides new insight into the structural basis of lipase specificities and a way to tune the substrate preference of lipases.
关键词：chain-length specificity • chimera • domain exchange