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亚洲中文字幕无线乱码:A variable loop involved in the substrate selectivity of pinoresinol/lariciresinol reductase from

作者:  文章来源:   点击数: 更新日期:2019-04-23
 

Title

A variable loop involved in the substrate selectivity of pinoresinol/

lariciresinol reductase from Camellia sinensis


Authors

Yingling Wua, Dawei Xinga, Guoliang Maa, Xinlong Daia, Liping Gaob??, Tao Xia?

 

Phytochemistry 162 (2019) 1–9


https://doi.org/10.1016/j.phytochem.2019.02.003.


Abstract

Pinoresinol/lariciresinol reductase (PLR), an NADPH-dependent reductase that catalyzes the sequential reduction of pinoresinol into secoisolariciresinol via Lariciresinol, can lead to the structural and stereochemical diversity of lignans. The relationship between substrate-selective reaction of PLR and sequence homology still remains unclear. In this study, we focused on the contribution of the variable region between PLRs in determining substrate selectivity. Here, two CsPLRs (CsPLR1 and CsPLR2) were identified in the tea plant (Camellia sinensis var. sinensis cv. Shuchazao). In vitro enzymatic assays showed that CsPLR1 could convert (+)- and (?)-pinoresinol into lariciresinol or secoisolariciresinol, whereas CsPLR2 catalyzed (+)-pinoresinol enantioselectively into (?)-secoisolariciresinol. Homology modeling and site-directed mutagenesis were used to examine the role of a variable loop in catalysis and substrate selectivity. The L174I mutant in CsPLR1 lost the capacity to reduce either (+)- or (?)-pinoresinol but retained the ability to catalyze the reduction of (?)-lariciresinol. These findings provide a basis for better understanding of the substrate-selective reaction of PLR.




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