吴伟鹏,于洪巍,叶丽丹,周萍萍,姚震,边旗

• 1:浙江大学化学工程与生物工程学院

摘要(Abstract)：

香叶基香叶醇(geranylgeraniol,GGOH)是生产香料、药物、维生素A和E的重要中间体,其生物活性构型为(E,E,E)-GGOH。为了提高酿酒酵母的(E,E,E)-GGOH产量,在表达了香叶基香叶基焦磷酸合酶-法尼基焦磷酸合酶融合酶(BTS1-DPP1)和香叶基香叶基焦磷酸合酶-二酰基甘油二磷酸磷酸酶融合酶(BTS1-ERG20)的初始菌株(产34.85mg?L~(-1)GGOH)的基础上,本实验进一步利用代谢工程的手段对法尼基焦磷酸(FPP)代谢节点处进行代谢流重排。一方面,过表达了来源于红法夫酵母的香叶基香叶基焦磷酸(GGPP)合酶突变体(Crt E03M),增加了前体物质GGPP的供应,从而将GGOH产量提高了1.6倍,达到56.04mg?L~(-1);另一方面,利用葡萄糖诱导性弱启动子PHXT1对竞争支路关键酶角鲨烯合酶(ERG9)进行下调,从而使GGOH产量提高4.5倍,达到156mg?L~(-1)。同时,通过敲除GAL80基因构建了葡萄糖调控的GAL体系,用以控制GGOH的合成,与PHXT1控制的角鲨烯合成支路一起,构成了顺序表达体系,最终,以十二烷为有机相,在摇瓶中进行两相发酵,培养72 h后获得GGOH产量183.07mg?L~(-1),与初始菌株相比提高了5倍。本工作对其他FPP下游途径的代谢调控具有借鉴意义。

关键词(KeyWords)： 香叶基香叶醇;代谢调控;顺序表达;葡萄糖响应;香叶基香叶基焦磷酸合酶

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(21576234,21776244)

作者(Author): 吴伟鹏,于洪巍,叶丽丹,周萍萍,姚震,边旗

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