摘要
利用大肠杆菌混菌发酵生物合成咖啡醇糖苷。通过在酪氨酸高产大肠杆菌BTAL中表达异源基因酪氨酸解氨酶(tyrosine ammonia-lyase from Rhodotorula glutinis,RgTAL),对羟基肉桂酰辅酶A连接酶(hydroxycinnamate:CoA ligase from Petroselinum crispum,Pc4CL)和肉桂酰辅酶A还原酶(cinnamyl-CoA reductase from Arabidopsis thaliana,AtCCR),与大肠杆菌BL21(DE3)中表达对羟基苯乙酸3-羟化酶HpaBC和糖基转移酶(UGT73C5 from A.thaliana,AtUGT73C5)混合培养发酵生产了3种咖啡醇葡萄糖苷,其中2种为非天然化合物。优化共培养比例至3∶1,咖啡醇-4-O-葡萄糖苷的最高产量达到(141.63±3.42)mg/L。这是首次微生物异源合成咖啡醇糖苷的报道,该工作扩大了咖啡醇糖苷的结构多样性,为后续的构效分析提供了新的候选化合物,有望得到具有较好生物活性的咖啡醇糖苷化合物。
Escherichia coli co-culture was designed for biosynthesis of caffeyl alcohol glucosides from glucose.Three caffeyl alcohol monoglucosides,namely caffeyl alcohol-3-O-glucoside,caffeyl alcohol-9-O-glucoside and caffeyl alcohol-4-O-glucoside,were produced by co-culturing two recombinant E.coli strains.A biosynthetic pathway of p-coumaryl alcohol was constructed in the high-tyrosine-producing E.coli BTAL-CAD01 by expressing tyrosine ammonialyase,hydroxycinnamate:CoA ligase(4CL),and cinnamyl-CoA reductase from Rhodotorula glutinis,Petroselinum crispum,and Arabidopsis thaliana,respectively,together with endogenous alcohol dehydrogenases or aldo-keto reductases.Then,4-hydroxyphenlacetate 3-hydroxylase(HpaBC)from E.coli and glucosyltransferase UGT73C5 from A.thaliana were co-expressed in wild-type E.coli BL21(DE3),resulting in a recombinant strain BWT-CAD01.Caffeyl alcohol monoglucosides were successfully produced by co-cultivating the recombinant strains BTAL-CAD01 and BWT-CAD01.The highest titer of caffeyl alcohol-4-O-glucoside reached(141.63±3.42)mg/L,with optimized inoculation ratio of 3∶1.To our knowledge,this is the first report on heterologous biosynthesis of caffeyl alcohol glucosides,both caffeyl alcohol-3-O-glucoside and caffeyl alcohol-9-O-glucoside are unnatural compounds.This work expands the structural diversity of caffeyl alcohol glucosides,which makes it possible the discovery of new analogues with improved pharmaceutical properties.
作者
王帅
庄以彬
刘浩
毕慧萍
刘涛
WANG Shuai;ZHUANG Yibin;LIU Hao;BI Huiping;LIU Tao(College of Engineering,Tianjin University of Science and Technology,Tianjin 300457,China;Tianjin Institute of Industrial Biotechnology,Chinese Academy of Sciences,Tianjin 300308,China)
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2020年第23期7-13,共7页
Food and Fermentation Industries
基金
国家自然科学基金项目(31970065)。
关键词
咖啡醇糖苷
生物合成
大肠杆菌
混菌培养
发酵生产
caffeyl alcohol glucosides
biosynthesis
Escherichia coli
microbial co-culture
fermentation production
作者简介
第一作者:王帅,硕士研究生;通讯作者:毕慧萍,副研究员,E-mail:bi_hp@tib.cas.cn。
