随机诱变和基因组重排选育阿维拉霉素高产菌  

Random mutagenesis and genome shuffling screen Streptomyces viridoehrongenes producing high-yield avilamycin

作　　者：胡向东 冯云会 叶茂 梁新乐[2] HU Xiangdong;FENG Yunhui;YE mao;LIANG Xinle(Hangzhou Crown Agricultural Biotechnology Co.Ltd,Hangzhou 310012,China;College of Food Science and Biotechnology,Zhejiang Gongshang University,Hangzhou 310018,China)

机构地区：[1]杭州皇冠农业生物工程技术研究中心有限公司,浙江杭州310012 [2]浙江工商大学食品与生物工程学院,浙江杭州310018

出　　处：《食品与发酵工业》2019年第7期1-7,共7页Food and Fermentation Industries

基　　金：浙江省重大科技专项重点农业项目(2015C02025)

摘　　要：以绿色产色链霉菌(Streptomyces viridoehrongenes 2-21)为原始出发菌株,依次进行常压等离子(atmospheric and room temperature, ARTP)和2轮^(137)Csγ诱变,从中筛选出高产阿维拉霉素的突变株,作为基因组重排的亲本菌株,以利福平为筛选剂,经过4轮原生质体递归融合,最终得到1株遗传稳定的高产阿维拉霉素重组菌株H_4-15。摇瓶发酵结果表明,阿维拉霉素产量可达到(2 155.48±7.81) mg/L,中试发酵结果表明,阿维拉霉素产量达到(2 356.44±6.34)mg/L,是原始出发菌株的3.47倍。高产菌株H_4-15发酵产物的LC-MS结果显示,其主要组分是阿维拉霉素A和阿维拉霉素B。综合表明,以利福平为筛选剂,结合传统随机诱变和基因组重排技术选育阿维拉霉素高产菌株,能大幅提升其生产能力,具有工业化生产的潜在价值。Streptomyces viridoehrongenes 2-21 was used as an original strain and treated with atmospheric room temperature plasma(ARTP) and two rounds of 137Csγ mutagenesis. Mutant strains with higher avilamycin productivity were screened and used as starting strains for genome shuffling. Using rifampicin as a selector, after four rounds of protoplast fusion, a high-yield-avilamycin-producing recombinant strain H4-15 was obtained. The avilamycin yield reached(2 155.48±7.81) mg/L in the flask, and(2 356.44±6.34) mg/L after a pilot-scale fermentation, which was 3.47 times higher than that of the original strain. Besides, its main components were found to be avilamycin A and avilamycin B. In conclusion, combining traditional random mutagenesis and genome shuffling and using rifampicin as a selector can significantly improve the yield of avilamycin, and its future potential value was proved.

关 键 词：绿色产色链霉菌 阿维拉霉素 随机诱变 基因组重排 

分 类 号：TQ920.1[轻工技术与工程—发酵工程]