Functional characterization of seven terpene synthases from Ophiopogon japonicus via engineered Escherichia coli  

作　　者：Wen Xu Qing Ma Tong Chen Haiyan Zhang Guanglei Ma Sibongile Mafu Juan Guo Xiaohui Fan Guanghong Cui Baolong Jin 

机构地区：[1]State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing,China [2]College of Pharmaceutical Sciences,Zhejiang University,Hangzhou,China [3]National Key Laboratory of Chinese Medicine Modernization,Innovation Center of Yangtze River Delta,Zhejiang University,Jiaxing,China [4]Department of Biochemistry and Molecular Biology,University of Massachusetts-Amherst,Amherst,MA,United States

出　　处：《Science of Traditional Chinese Medicine》2024年第4期312-318,共7页中医药科学（英文）

基　　金：supported by National Natural Science Foundation of China(U23A20513);the Fundamental Research Funds for the Central Public Welfare Research Institutes(ZZ14-YQ-049);National Natural Science Foundation of China(82274054);Key project at central government level:the ability to establish sustainable use of valuable Chinese Medicine Resources(2060302).

摘　　要：Background:Ophiopogon japonicus(L.f)Ker-Gawl.growing in Zhejiang is recognized as the Dao-di medicinal herb for the production of Ophiopogonis Radix.Borneol-7-O-[β-D-apiofuranosyl-(1→6)]-β-D-glucopyranoside,a prominent pharmacologically active compound,serves as a marker distinguishing O.japonicus in Zhejiang from those in other geographical areas.It is synthesized from borneol through glycosylation,with terpene synthase(TPS)being the critical enzyme catalyzing the conversion of terpene precursors into borneol.Objective:The aim of the study was to define key genes involved in biosynthesis of borneol in O.japonicus.Methods:The candidate terpene synthase genes were identified from the root and leaf transcriptome data of O.japonicus in Zhejiang and the functions of these enzymes were identified using engineered Escherichia coli.Results:This study developed a rapid expression system for monoterpene and sesquiterpene synthases based on engineered E.coli.Seven terpene synthase genes(OjTPS1 to OjTPS7)were identified in different terpene synthase subfamilies,including 2 from TPS-a,4 from TPS-b,and 1 from TPS-g.Biochemical analysis using an engineered system E.coli demonstrated that all the 7 terpene synthases produced monoterpenes,and OjTPS3,OjTPS5,and OjTPS6 also yielded sesquiterpenes.Conclusions:These 7 terpene synthases produced 13 monoterpenes and 8 sesquiterpenes.Notably,OjTPS1 produced borneol establishing the groundwork for elucidating the biosynthetic pathways of borneol-7-O-[β-D-apiofuranosyl-(1→6)]-β-D-glucopyranoside and other volatile oil components.

关 键 词：Ophiopogon japonicus Ophiopogon japonicus in Zhejiang Terpene synthase BORNEOL 

分 类 号：O62[理学—有机化学]