Genetic Cloning and Functional Verification of Hydroxycinnamoyltransferase in Ixeris sonchifolia Hance

doi:10.13560/j.cnki.biotech.bull.1985.2018-0340

Abstract

Abstract: Phenolic acid compounds from Ixeris sonchifolia Hance,such as chlorogenic acid,etc.,are reported to have antioxidation and free radicals scavenging activities. Hydroxycinnamoyltransferase（HCT）is a key enzyme in the biosynthetic pathway of plant chlorogenic acid. A hydroxycinnamoyltransferase gene from I. sonchifolia Hance leaves was cloned using transcriptome sequencing,sequence analysis and RT-PCR strategies,and named as IsHCT（GenBank accession number：MH151086）. The open read frame of the gene was 1320 bp encoding a putative protein of 439 amino acids. Multiple sequence alignment and bioinformatics analysis results showed that IsHCT contained two conserved domains,and belonged to the HCT enzyme in the BAHD family. Phylogenetic tree analysis demonstrated that this gene had the highest homology with CiHCT from Cichorium intybus,and the similarity reached 95%.Finally,by overexpressing IsHCT and At4CL from Arabidopsis thaliana in Escherichia coli and feeding the substrates caffeic acid and quinic acid in the fermentation medium,and chlorogenic acid was detected in the fermentation liquid,indicating that the IsHCT catalyzed the chlorogenic acid synthesis from caffeyl coenzyme A and quinic acid,i.e.,it functioned as HCT

Key words: Ixeris sonchifolia Hance, HCT, chlorogenic acid, Escherichia coli

LI Tian-zhen, ZHOU Wei, YIN Hua, ZHANG Tong-cun, LIU Tao. Genetic Cloning and Functional Verification of Hydroxycinnamoyltransferase in Ixeris sonchifolia Hance[J]. Biotechnology Bulletin, 2018, 34(11): 120-126.

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