酿酒酵母过表达Ehrlich途径基因高效合成色醇  

Efficient synthesis of indole-3-ethanol in Saccharomyces cerevisiae byoverexpression of Ehrlich pathway genes

作　　者：孙井震 何玙冰 杨卫华 杨艳坤[1,2] 刘秀霞 刘春立[1,2] 詹锦铃 李业 白仲虎 SUN Jingzhen;HE Yubing;YANG Weihua;YANG Yankun;LIU Xiuxia;LIU Chunli;ZHAN Jinling;LI Ye;BAI Zhonghu(School of Biotechnology,Jiangnan University,Wuxi 214122,China;National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing,Jiangnan University,Wuxi 214122,China;Changxing Pharmaceutical Co.Ltd.,Changxing 313100,China)

机构地区：[1]江南大学生物工程学院,江苏无锡214122 [2]江南大学,粮食发酵与食品生物制造国家工程研究中心,江苏无锡214122 [3]长兴制药股份有限公司,浙江长兴313100

出　　处：《食品与发酵工业》2022年第24期16-23,共8页Food and Fermentation Industries

基　　金：国家自然科学基金面上项目(21878124)。

摘　　要：色醇(indole-3-ethanol,IET)正在日益成为重要的医药和化工合成的中间体,但是其合成目前还是以化学合成为主。该文通过对酿酒酵母(Saccharomyces cerevisiae)Ehrlich途径相关基因的研究,利用不同基因的组合,通过生物生产IET来解决化学合成不环保、成本高的问题。该研究对涉及色氨酸代谢到IET的代谢途径中相关的13个基因根据中间产物吲哚丙酮酸和吲哚乙醛的代谢过程进行了分步骤的过表达,运用高效液相色谱对发酵液进行产物的检测,同时检测竞争途径副产物对羟基苯乙醇和苯乙醇的变化,探索出了最适合的利于IET生物生产的基因组合,并探究出了该过程的主要限速步骤,使IET在添加5 mmol/L色氨酸情况下得到了1.09 mmol/L的产量。Indole-3-ethanol(IET)is a metabolic derivative of tryptophan(Trp)that exhibits pleiotropic effects in humans,plants and microorganisms.The mechanism of IET biosynthesis was originally explored decades ago.IET is an essential intermediate for pharmaceutical and chemical synthesis,but its industrial production is dominated by chemical synthesis.In this paper,we investigated genes related to the Ehrlich pathway of Saccharomyces cerevisiae to solve the problem of environmentally unfriendly and costly chemical synthesis by combining different genes for biological production IET.In this study,13 genes related to the metabolic pathway involving the metabolism of tryptophan(Trp)to IET were overexpressed in steps according to the metabolic processes of the intermediate products indole-3-pyruvate(IPY)and indole-3-acetaldehyde(IAC),and the fermentation broth was examined for the products using HPLC,as well as for changes in by-products of competing pathways p-hydroxy-phenylethanol(pPET)and phenylethanol(PET).In order to observe the changes of various substances related to IET production more intuitively and conveniently,the process of biological production was further explored by adding Trp.At the same time,the remaining amount of Trp after fermentation was detected,and the relationship between the amount of endogenous and exogenous Trp in IET production was simply explored.In addition,the most suitable gene combinations for IET bioproduction were explored,and the main rate-limiting steps of the process were explored,resulting in a yield of 1.09 mmol/L of IET with the addition of 5 mmol/L Trp.

关 键 词：色醇 色氨酸 生物生产 酿酒酵母 Ehrlich途径 

分 类 号：TQ923[轻工技术与工程—发酵工程]