常压室温等离子体与5-溴尿嘧啶复合诱变及快速选育腺苷高产菌株  

Combined mutagenesis of ARTP and 5-BU for improving production of adenosine in Bacillus subtilis

作　　者：杨心萍 宋词 张伟豪 刘艳 王洲 薛正莲[1,2] YANG Xinping;SONG Ci;ZHANG Weihao;LIU Yan;WANG Zhou;XUE Zhenglian(College of Biological and Chemical Engineering,Anhui Polytechnic University,Wuhu 241000,China;Anhui Engineering Technology Research Center of Microbial Fermentation,Wuhu 241000,China)

机构地区：[1]安徽工程大学生物与化学工程学院,安徽芜湖241000 [2]微生物发酵安徽省工程技术研究中心,安徽芜湖241000

出　　处：《食品与发酵工业》2020年第9期73-77,共5页Food and Fermentation Industries

摘　　要：采用常压室温等离子体(atmospheric room temperature plasma, ARTP)与5-溴尿嘧啶(5-bromouracil, 5-BU)对枯草芽孢杆菌B-0进行复合诱变,通过磺胺胍(sulfaguanidine, SG)抗性初筛、48孔板定量复筛的培养方法快速选育腺苷高产菌株。结果显示,48孔板在枯草芽孢杆菌发酵过程中存在边缘孔效应,并应用2 mL无菌水填充可消除边缘孔效应,且通过6轮复合诱变最终获得1株较出发菌株腺苷产量提升35%且遗传稳定的突变株B-6。ARTP与5-BU复合诱变结合48孔板快速选育,显著提高腺苷高产菌株选育效率与选育的准确率,也为其他核苷类菌株的选育提供了理论参考。The mutagenesis of Bacillus subtilis B-0 was carried out using atmospheric and room temperature plasma(ARTP) and 5-bromouracil(5-BU), and the adenosine high-yield strain was selected by sulfaguanidine(SG) resistance and in 48-well plates. A genetic stable mutant strain B-6 with 35% higher yield of adenosine than that of Bacillus subtilis B-0 was finally obtained after 6 cycles of mutagenesis and screening. Briefly, the combination of ARTP and 5-BU mutagenesis combined with 48-well plates can significantly improve the breeding efficiency and accuracy of adenosine high-yielding strains, and also provide a theoretical reference for the selection of other nucleoside strains.

关 键 词：腺苷 枯草芽孢杆菌 常压室温等离子体 化学诱变 微孔板培养 

分 类 号：TQ929[轻工技术与工程—发酵工程]