3-(4-羟基苯基)丙酸在酿酒酵母中的生物合成  

Biosynthesis of 3-(4-Hydroxyphenyl)Propionic Acid in Saccharomyces cerevisiae

作　　者：江晶洁 庄以彬[2] 殷华[2] 刘涛[2] 刘少伟[1] JIANG Jlng-jle ZHUANG Yi-bin YIN Hua LIU Tao LIU Shao-wei(College of Biotechnology , the State Key Laboratory of Bioreactor Engineering , East China University of Scienee and Technology , Shanghai 200237 Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308)

机构地区：[1]华东理工大学,上海200237 [2]中国科学院天津工业生物技术研究所,天津300308

出　　处：《生物技术通报》2017年第9期184-190,共7页Biotechnology Bulletin

基　　金：国家自然科学基金资助项目(31400026,21302214);国家重点基础研究发展计划("973"计划)(2012CB721100)

摘　　要：3-(4-羟基苯基)丙酸(HPPA)是一种芳香类化合物,作为中间体主要应用于医药、食品、化学领域,具有重要的经济价值。旨在实现HPPA在酿酒酵母中的从头合成,通过在酿酒酵母中过表达约氏黄杆菌来源的酪氨酸转移酶(Fj TAL)、拟南芥来源的对香豆酰辅酶A连接酶(At4CL),同时利用酿酒酵母BY4742自身来源的超长链烯酰辅酶A还原酶(Sc TSC13)、硫酯水解酶,实现了在酿酒酵母中从头合成HPPA,产量约为70 mg/L左右。在以4 mmol/L酪氨酸为前体、半乳糖诱导Fj TAL过表达、30℃发酵培养72 h的条件下,约36%的酪氨酸转化生成对香豆酸;在以4 mmol/L对香豆酸或咖啡酸为前体,半乳糖诱导At4CL过表达,同时利用酵母内源Sc TSC13、硫酯水解酶,30℃发酵培养72 h的条件下,约94%的对香豆酸被还原成HPPA,约91%的咖啡酸被还原成3,4-二羟基苯丙酸(DHPPA),为进一步生物合成HPPA及其衍生物奠定了基础。3-（4-Hydroxyphenyl）propionic acid（HPPA）is an aromatic compound with importantly economic value,because itis an intermediate mainly used in medicine,food and chemical industry. In this work,we aim to achieve the De novo synthesis of HPPA inSaccharomyces cerevisiae,and it was conducted via overexpressing a tyrosine ammonia-lyase from Flavobacterium johnsoniaeu（Fj TAL）anda p-coumaroyl-Co A ligase from Arabidopsis thaliana（At4CL）,as well as using super long-chain enoyl-Co A reductase（Sc TSC13）andacyl-Co A thioesterases from S. cerevisiae BY4742. The yield of HPPA reached 70 mg/L. Bioconversion rate of tyrosine into p-coumaric acidwas 36% under the condition of 4 mmol/L tyrosine enhancement in shake flask after 72 h fermentation at 30℃ and galactose inducing Fj TALoverexpression. While in the recombinant S. cerevisiae overexpressing At4 CL combined with enzymes Sc TSC13 and acyl-Co A thioesterases,94% of 4 mmol/L p-coumaric acid was converted to HPPA in shake flask after 72 h fermentation at 30℃,and 91 % of 4 m M caffeic acid wasconverted to DHPPA. This work laid a foundation for the biosynthesis of HPPA and its derivatives.

关 键 词：酿酒酵母 3-(4-羟基苯基)丙酸(HPPA) 生物合成 酪氨酸氨基转移酶 对香豆酰辅酶A连接酶 

分 类 号：Q936[生物学—微生物学]