Functional characterization of triterpene synthases in Cibotium barometz  

作　　者：Zhongju Ji Baolian Fan Yidu Chen Jingyang Yue Jiabo Chen Rongrong Zhang Yi Tong Zhongqiu Liu Jincai Liang Lixin Duan 

机构地区：[1]Guangdong Provincial Key Laboratory of Translational Cancer Research of Chinese Medicines,Joint International Research Laboratory of Translational Cancer Research of Chinese Medicines,International Institute for Translational Chinese Medicine,School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou,China

出　　处：《Synthetic and Systems Biotechnology》2023年第3期437-444,共8页合成和系统生物技术（英文）

基　　金：the National Natural Science Foundation of China(No.81874333);the Key Laboratory of Guangdong Drug Administration(2021ZDB03);the Guangdong Basic and Applied Basic Research Foundation(No.2020B1515130005).

摘　　要：Cibotium barometz(Linn.)J.Sm.,a tree fern in the Dicksoniaceae family,is an economically important industrial exported plant in China and widely used in Traditional Chinese Medicine.C.barometz produces a range of bioactive triterpenes and their metabolites.However,the biosynthetic pathway of triterpenes in C.barometz remains unknown.To clarify the origin of diverse triterpenes in C.barometz,we conducted de novo transcriptome sequencing and analysis of C.barometz rhizomes and leaves to identify the candidate genes involved in C.barometz triterpene biosynthesis.Three C.barometz triterpene synthases(CbTSs)candidate genes were obtained.All of them were highly expressed in C.barometz rhizomes,consisting of the accumulation pattern of triterpenes in C.barometz.To characterize the function of these CbTSs,we constructed a squalene-and oxidosqualene-overproducing yeast chassis by overexpressing all the enzymes in the MVA pathway under the control of GAL-regulated promoter and disrupted the GAL80 gene in Saccharomyces cerevisiae simultaneously.Heterologous expressing CbTS1,CbTS2,and CbTS3 in engineering yeast strain produced cycloartenol,dammaradiene,and diploptene,respectively.Phylogenetic analysis revealed that CbTS1 belongs to oxidosqualene cyclase,while CbTS2 and CbTS3 belong to squalene cyclase.These results decipher enzymatic mechanisms underlying the origin of diverse triterpene in C.barometz.

关 键 词：2 3-oxidosqualene cyclase Squalene cyclase Cibotium barometz Triterpene biosynthesis 

分 类 号：O62[理学—有机化学]