利用橡胶树来源β-石竹烯新合成酶构建工程酿酒酵母的研究  

Construction of engineering Saccharomyces cerevisiae with a newβ-caryophyllene synthase from Hevea brasiliensis

作　　者：石彬 娄晶 杨竹如 邓小敏[2] SHI Bin;LOU Jing;YANG Zhuru;DENG Xiaomin(College of Life and Health,Wuhan Vocational College of Software and Engineering(Wuhan Open University),Wuhan 430205,China;Key Laboratory of Biology and Genetic Resources of Rubber Tree of Ministry of Agriculture and Rural Affairs,State Key Laboratory Incubation Base for Cultivation&Physiology of Tropical Crops,Rubber Research Institute,Chinese Academy of Tropical Agricultural Sciences,Haikou 571101,China)

机构地区：[1]武汉软件工程职业学院(武汉开放大学)生命健康学院,湖北武汉430205 [2]中国热带农业科学院橡胶研究所,农业农村部橡胶树生物学与遗传资源利用重点实验室,海南省热带作物栽培生理学重点实验室,海南海口571101

出　　处：《中国酿造》2023年第8期147-152,共6页China Brewing

基　　金：武汉市知识创新项目曙光计划(2022022020801020440);海南省自然科学基金高层次人才项目(320RC734);海南省自然科学基金(323QN298)。

摘　　要：为了利用生物技术高效合成β-石竹烯,该研究从橡胶树(Hevea brasiliensis)中筛选并鉴定到β-石竹烯新合成酶,并构建高产β-石竹烯酿酒酵母(Saccharomyces cerevisiae)工程菌株。结果表明,在橡胶树中筛选鉴定出一个新的β-石竹烯合成酶基因,命名为HbBACS,其与已报道的β-石竹烯合成酶相比,编码蛋白的氨基酸序列同源性和相似性均较低。利用密码子优化的该基因构建工程酿酒酵母YCAR02,其β-石竹烯摇瓶发酵产量为206 mg/L。进一步对工程酿酒酵母YCAR02进行两阶段分批补料发酵,在添加10%肉豆蔻酸异丙酯覆盖剂的条件下,5 L发酵罐中β-石竹烯产量达到8.47 g/L。橡胶树来源的新β-石竹烯合成酶活性较高,这为利用合成生物技术规模化生产β-石竹烯提供了新的思路与方法。To efficiently synthesizeβ-caryophyllene using biotechnology,a newβ-caryophyllene synthase was screened and identified from Hevea brasiliensis,and the engineering strain of Saccharomyces cerevisiae with highβ-caryophyllene yield was constructed.The results showed that a newβ-caryophyllene synthase gene,named as HbBACS,was screened and identified in H.brasiliensis.Compared with the reportedβ-caryophyllene synthase,the homology and similarity of amino acid sequences encoding protein were lower.The engineering S.cerevisiae YCAR02 was constructed with the codon-optimized gene and theβ-caryophyllene yield was 206 mg/L in shake flask fermentation.Further,the two-stage batch feed fermentation of engineering S.cerevisiae YCAR02 was carried out.Under the condition of adding 10%isopropyl myristate coating agent,theβ-caryophylene yield in 5 L fermenter reached 8.47 g/L.The activity ofβ-caryophyllene synthetase from H.brasiliensis was higher,which provided a new idea and method for large scale production ofβ-caryophyllene by synthetic biotechnology.

关 键 词：β-石竹烯 橡胶树 新合成酶 酿酒酵母 倍半萜 分批补料发酵 

分 类 号：Q819[生物学—生物工程] TS201.2[轻工技术与工程—食品科学]