收稿日期: 2012-03-29
修回日期: 2012-04-13
网络出版日期: 2012-04-13
基金资助
国家自然科学基金(61173098)和中国科学院"十二五"基础前沿专项(KSCX2-EW-J-29)资助
Cloning and characterization of PcCHS1 from Polygonum cuspidatum
Received date: 2012-03-29
Revised date: 2012-04-13
Online published: 2012-04-13
李星 , 王红 . 虎杖查尔酮合酶PcCHS1基因的克隆与功能分析[J]. 中国科学院大学学报, 2013 , 30(2) : 206 -212 . DOI: 10.7523/j.issn.1002-1175.2013.02.010
A full length cDNA encoding a type Ⅲ PKS (PcCHS1) was isolated from Polygonum cuspidatum Sieb. et Zucc. Functional and enzymatic assays biochemically confirmed that PcCHS1 was a chalcone synthase. When incubated with p-coumaroyl-CoA and malonyl-CoA at pH 7-8, PcCHS1 catalyzed the formation of chalcone as the major product. However, at pH 9 both p-hydroxybenzalacetone and chalcone were detected. Site-directed mutagenesis of Phe216 into Leu caused reduction of enzyme activity. Another mutation N333K performed activity of releasing some common derailment products, CTAL and BNY, of many CHSs in vitro.
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