高能燃料前体α-红没药烯的生物制造过程优化  

Optimization of biomanufacturing process of high-energy fuel precursorα-bisabolene

作　　者：马治东 张亚鹏 高慧鹏 李文强 吕波 秦磊 张全 李春[1,2,4] MA Zhidong;ZHANG Yapeng;GAO Huipeng;LI Wenqiang;LYU Bo;QIN Lei;ZHANG Quan;LI Chun(School of Chemistry and Chemical Engineering/State Key Laboratory Incubation Base for Green Processing of Chemical Engineering,Shihezi University,Shihezi 832003,Xinjiang,China;Key Laboratory of Medical Molecule Science and Pharmaceutics Engineering,Ministry of Industry and Information Technology,Institute of Biochemical Engineering,School of Chemistry and Chemical Engineering,Beijing Institute of Technology,Beijing 100081,China;Sinopec Dalian Research Institute of Petroleum and Petrochemicals,Dalian 116023,Liaoninng,China;Key Laboratory for Industrial Biocatalysis,Ministry of Education,Department of Chemical Engineering,Tsinghua University,Beijing 100084,China)

机构地区：[1]石河子大学化学化工学院/化工绿色过程省部共建国家重点实验室培育基地,新疆石河子832003 [2]北京理工大学化学与化工学院生物化工研究所/医药分子科学与制剂工信部重点实验室,北京100081 [3]中石化大连石油化工研究院有限公司,辽宁大连116023 [4]清华大学化学工程系工业生物催化教育部重点实验室,北京100084

出　　处：《化工学报》2024年第12期4702-4711,F0004,共11页CIESC Journal

基　　金：国家自然科学基金项目(22138006);中国石化课题(421127)。

摘　　要：α-红没药烯是一种主要存在于红没药、柠檬等植物精油中的倍半萜化合物,因具备多支链、环状结构,其烷化产物红没药烷可作为航空燃料替代品,受到了国内外的广泛关注。以酿酒酵母为底盘细胞来合成α-红没药烯是一种绿色、可持续的备选生产方式。本研究在强化甲羟戊酸(mevalonate,MVA)合成途径的酿酒酵母中表达来自北美冷杉的α-红没药烯合酶AgBIS,利用葡萄糖诱导型启动子PHXT1对支路鲨烯合酶及脂肪酸合成途径进行动态下调,进一步系统增强MVA途径多个基因表达,并对辅因子供应与乙醇利用途径进行强化,使α-红没药烯在摇瓶中的产量达到510 mg/L。在3 L发酵罐中,通过乙醇补料发酵使α-红没药烯产量达到16.5 g/L,为目前报道的最高产量。本研究为微生物细胞工厂高效合成α-红没药烯提供了有效策略。α-bisabolene is a sesquiterpene compound mainly found in plant essential oils such as myrrh and lemon.Due to its multi-branched and cyclic structure,its alkylation product bisabolane can be used as a substitute for aviation fuel and has attracted widespread attention at home and abroad.The biosynthesis ofα-bisabolene using Saccharomyces cerevisiae as a chassis is a green and sustainable alternative production mode.In this study,aα-bisabolene synthase AgBIS from Abies grandis was expressed in S.cerevisiae with the enhanced key genes of the mevalonate(MVA)pathway.The glucose inducible promoter PHXT1 was used to dynamically downregulate the endogenous squalene synthase and fatty acid synthesis pathway.Further systematic enhancement of the expression of multiple genes in the MVA pathway,cofactor supplementary and ethanol utilization pathway systematically,theα-bisabolene production reached 510 mg/L in the flask.In a 3 L fermenter,the highest reported production ofα-bisabolene was achieved at 120 h with ethanol supplementation,reaching 16.5 g/L.This study provides an effective strategy and research foundation for the efficient biosynthesis ofα-bisabolene by microbial cell factories.

关 键 词：α-红没药烯 酿酒酵母 代谢工程 微生物细胞工厂 合成生物学 

分 类 号：Q819[生物学—生物工程]