Molecular Cloning and Bacterial Expression of Germacrene A Synthase cDNA from Crepidiastrum sonchifolium

作     者：REN Jie LIU Yan-qiu YANG Li NI Rui YOU Song 

作者机构：School of Pharmaceutical Engineering Shenyang Pharmaceutical UniversityShenyang 110016 P. R. China 

出 版 物：《Chemical Research in Chinese Universities》 (高等学校化学研究（英文版）)

年 卷 期：2006年第22卷第5期

页      面：606-611页

核心收录：

学科分类：1008[医学-中药学(可授医学、理学学位)] 1006[医学-中西医结合] 0703[理学-化学] 100602[医学-中西医结合临床] 10[医学] 

主　　题：Germacrene A synthase Crepidiastrum sonchifolium Sesquiterpene synthase Expression Fusion protein 

摘      要：Germacrene A synthase（GAS） catalyzes the biosynthesis of germacrene A, which is a key precursor for sesquiterpone lactones. Cloning of a novel full-length cDNA encoding GAS from the medicinal plant Crepidiastrum sonchifolium（dosignated CsGAS） is reported in this study. The cDNA is 1837 bp long and contains a 1680-bp open reading frame encoding a 559 amino-acid protein. The functional expression of the cDNA in Escherichia coli, as an N-terminal thioredoxin fusion protein, with the pET32a vector yielding a recombinant enzyme. Sequence analysis was used to compare this enzyme with the mechanistically related epi-aristolochene synthase from tobacco, and the effect of possible involvement of a number of amino acids in sesquiterpone synthase on product specificity was also discussed.