产4-乙烯基愈创木酚细菌的筛选及酚酸脱羧酶基因的克隆与表达  

Screening of 4-vinylguaiacol-producing bacteria and cloning and expression of phenolic acid decarboxylase gene

作　　者：巩园园 毛豪 晋湘宜 冯向东 陈茂彬 方尚玲 GONG Yuanyuan;MAO Hao;JIN Xiangyi;FENG Xiangdong;CHEN Maobin;FANG Shangling(Key Laboratory of Fermentation Engineering,Ministry of Education,School of Food and Biological Engineering,Hubei University of Technology,Wuhan 430068,China;Hubei Daohuaxiang Wine Industry Co.,Ltd.,Yichang 443000,China)

机构地区：[1]湖北工业大学生物工程与食品学院教育部发酵工程重点实验室,湖北武汉430068 [2]湖北稻花香酒业股份有限公司,湖北宜昌443000

出　　处：《中国酿造》2021年第7期160-164,共5页China Brewing

基　　金：“十三五”国家重点研发计划项目(2016YFD0400500);湖北省科技厅重大专项(2018ABA084)。

摘　　要：采用传统筛菌方法从发酵醅中筛选产4-乙烯基愈创木酚(4-VG)的细菌,通过形态观察及分子生物学技术对其进行菌种鉴定,并采用基因克隆技术对其酚酸脱羧酶基因进行克隆与异源表达。结果表明,筛选得到一株产4-VG的菌株NF1,其4-VG产量为0.30 mg/g,并鉴定该菌株为枯草芽孢杆菌(Bacillus subtilis)。从该菌株中成功克隆得到酚酸脱羧酶基因,并在大肠杆菌(Escherichia coli)表达系统中进行诱导表达,以50 mmol/L的阿魏酸为底物,酚酸脱羧酶酶活力为66.74 IU/mL,经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)检测,该酶的分子质量约为20 kDa。A 4-vinylguaiacol(4-VG)producing bacterium was screened from fermented grains by traditional screening method,and identified through morphological observation and molecular biology technology.The phenolic acid decarboxylase gene was cloned and expressed heterologously through gene cloning technology.The results showed that a 4-VG-producing strain NF1 was screened and identified as Bacillus subtilis,and its 4-VG yield was 0.30 mg/g.The phenolic acid decarboxylase gene was successfully cloned from this strain and expressed in an E.coli expression system.With ferulic acid 50 mmol/L as the substrate,the activity of the phenolic acid decarboxylase was 66.74 IU/ml.Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)results showed that the molecular mass of the enzyme was 20 kDa.

关 键 词：4-乙烯基愈创木酚 筛选 鉴定 酚酸脱羧酶 克隆 表达 

分 类 号：Q786[生物学—分子生物学]