咖啡酸苯乙酯生物合成体系的构建及优化  

Construction and optimization of Caffeic acid phenethyl ester biosynthesis pathway

作　　者：张根林[1] 李媛媛 邱笛 赵金雨 ZHANG Genlin;LI Yuanyuan;QIU Di;ZHAO JinYu(College of Chemistry and Chemical Engineering/Key Laboratory of Chemical Green Processing of Xinjiang Corps,Shihezi University,Shihezi 832003,Xinjiang,China)

机构地区：[1]石河子大学化学化工学院/化工绿色过程兵团重点实验室,新疆石河子832003

出　　处：《西安工程大学学报》2025年第1期9-16,共8页Journal of Xi’an Polytechnic University

基　　金：国家自然科学基金(22178226)。

摘　　要：咖啡酸苯乙酯具有多种药理活性,但天然来源受限,化学合成存在环境隐患,利用合成生物技术开发咖啡酸苯乙酯的微生物合成更符合绿色经济要求。通过构建咖啡酸苯乙酯从头合成途径、强化酰基转移酶表达、优化2-苯乙醇途径和苯丙酮酸途径等策略,实现咖啡酸苯乙酯的酵母合成,酵母工程菌S4发酵120 h后咖啡酸苯乙酯产量达到(98.56±4.56)μg/L。平衡2种前体咖啡酰辅酶A和2-苯乙醇供应,在大肠杆菌中表达酪氨酸解氨酶(TAL)、羟基肉桂酰基转移酶(HCT)和辅酶A连接酶(4CL)形成咖啡酰辅酶A合成工程菌强化咖啡酰辅酶A供应,与酵母工程菌S4构建混菌发酵体系,混菌培养条件优化后咖啡酸苯乙酯产量达到(667.64±10.26)μg/L。Caffeic acid phenethyl ester(CAPE)has various pharmacological activities;however,its natural sources are limited,and chemical synthesis poses environmental hazards.Its microbial synthesis using synthetic biotechnology will be more in line with green economy.In this study,CAPE was synthesized in yeast by constructing its de novo synthesis pathway,overexpressing acyltransferase,optimizing 2-phenylethanol pathway and phenylpyruvate pathway.After 120 h of fermentation,the titer of it reached(98.56±4.56)μg/L by yeast engineering strain S4.After the balancing of the supply of two precursors,caffeoyl coenzyme A and 2-phenylethanol,a caffeoyl CoA synthesis engineering strain was consrtruted through expressing tyrosine deaminase(TAL),hydroxycinnamoyl transferase(HCT)and CoA ligase(4CL)in Escherichia coli.Then,a mixed culture system was constructed with yeast engineering strain S4 and caffeoyl CoA synthesis engineering strain,resulted in(667.64±10.26)μg/L of CAPE after optimizing the culture conditions.This study provides a reference for the cellular engineering synthesis of CAPE.

关 键 词：咖啡酸苯乙酯 途径优化 生物合成 混菌发酵 2-苯乙醇 

分 类 号：Q815[生物学—生物工程]