高产肌醇-1-磷酸合酶大肠杆菌重组工程菌的构建及其产肌醇发酵条件优化  

Construction of high-yield inositol-1-phosphate synthase recombinant Escherichia coli andoptimization of fermentation conditions for myo-inositol production

作　　者：易明花 任国平 胡琼 钱蒙蒙 YI Minghua;REN Guoping;HU Qiong;QIAN Mengmeng(Department of Health and Tourism,Hangzhou Wanxiang Polytechnic,Hangzhou 310014,China;Research Center of Good Hangzhou,Hangzhou Wanxiang Polytechnic,Hangzhou 310014,China)

机构地区：[1]杭州市万向职业技术学院康养旅游系,浙江杭州310014 [2]杭州市万向职业技术学院美好杭州研究中心,浙江杭州310014

出　　处：《中国酿造》2024年第12期143-148,共6页China Brewing

基　　金：浙江省教育厅项目(232022020);浙江省新苗计划项目(2023R449);杭州万向职业技术学院项目(YN2023048)。

摘　　要：该研究以酿酒酵母(Saccharomyces cerevisiae)SC288基因组为模版,利用基因工程技术克隆肌醇合成途径关键酶肌醇-1-磷酸合酶(IPS)的表达基因INO1,以质粒pETDuet-1为载体,构建重组质粒pETDuet-INO1,导入大肠杆菌(Escherichia coli)BL21,构建重组菌株E.coli BL21/pETDuet-INO1,实现IPS在大肠杆菌高效表达,并采用单因素试验及正交试验对其产肌醇的发酵条件进行优化。结果表明,成功构建重组大肠杆菌E.coli BL21/pETDuet-INO1,其发酵产肌醇的最佳工艺条件为:初始葡萄糖添加量12 g/L,诱导温度28℃,接种量3%。在此条件下进行诱导发酵24 h,肌醇产量高达(0.81±0.007)g/L,比优化前提高60.39%。In this study,the recombinant plasmid pETDuet-INO1 was constructed by introducing the expression gene INO1 of inositol-1-phosphate synthase(IPS)(a key enzyme in inositol synthesis pathway)cloned from Saccharomyces cerevisiae SC288 by gene engineering technology into plasmid pETDuet-1.The recombinant strain Escherichia coli BL21/pETDuet-INO1 was constructed by introducing the recombinant plasmid pETDuet-INO1 into E.coli BL21 to achieve efficient expression of IPS in E.coli.The fermentation conditions for myo-inositol production were optimized by single factor experiments and orthogonal experiments.The results showed that the recombinant E.coli BL21/pETDuet-INO1 was successfully constructed,and the optimal fermentation conditions for myo-inositol production were as follows:initial glucose addition 12 g/L,induction temperature 28℃,and inoculum 3%.The yield of myo-inositol was(0.81±0.007)g/L after induction 24 h under these conditions,which was 60.39%higher than that before optimization.

关 键 词：大肠杆菌 肌醇-1-磷酸合酶 基因工程 肌醇 正交试验 发酵条件 

分 类 号：Q812[生物学—生物工程]