桂花萜烯合酶（TPS）的生物信息学与原核表达分析

文章编号：1000-3142（2019）05-0624-09

Bioinformatic and prokaryotic expression analysis ofterpene synthase （TPS） from Osmanthus fragrans

ZENG Xiangling1，2， ZOU Jingjing1，2， WANG Caiyun2*

（ 1. School of Nuclear Technology and Chemistry & Biology， Hubei University of Science and Technology， Xianning 437100， Hubei， China;2. Key Laboratory for Biology of Horticultural Plants， Ministry of Education， Huazhong Agricultural University， Wuhan 430070， China ）

Abstract：Volatile terpenes are important aroma active compounds in flowers and fruits of plant. Diversities of terpenes are usually determined by type and function of terpene synthase in different species. Osmanthus fragransis an important fragrant plant， in which terpenes are the important components of floral scent. But there are few studies on terpene synthase in O. fragrans. To reveal the biosynthesis mechanism of terpenes inO. fragrans，we predicted the physicochemical properties， subcellular localization and structure of four TPS proteins by bioinformatics， and expressed them in prokaryotic expression system. Finally， the function of soluble TPS4 recombinant protein was analyzed by enzyme reaction in vitro. The results were as follows： （1） The physicochemical properties of the four TPS proteins had relatively little difference. Only TPS4 protein locating in other targets without signal peptide had low proportional extended strand and no extended strand near amidogen terminal. （2） All of the four TPS proteins were successfully expressed in prokaryotic system， but only TPS4 obtained soluble recombinant protein. （3） The purified TPS4 recombinant protein was reacted with GPP， NDP and FPP respectively， and only one product （trans-β-ocimene， β-phellandrene and α-farnesene） was detected. The results provide reference for functional analysis of floral scent related gene at protein level in O. fragransand for revealing the molecular mechanism of terpenes biosynthesis in plant.