Cloning and characterization of PcCHS1 from Polygonum cuspidatum

doi:10.7523/j.issn.1002-1175.2013.02.010

Abstract

Abstract:

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.

Key words: Polygonum cuspidatum, chalcone synthase, site-directed mutagenesis, enzyme activity

CLC Number:

S567.239

LI Xing, WANG Hong. Cloning and characterization of PcCHS1 from Polygonum cuspidatum[J]. , 2013, 30(2): 206-212.

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