利用抗性筛选和基因组重排技术选育ε-聚赖氨酸高产菌株

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英文篇名：Improving ε-poly-L-lysine productivity in Streptomyces albulus M-Z18 through antibiotic resistance screening and genome shuffling technology 作者：赵俊杰 ; 王开方 ; 陈旭升 ; 毛忠贵 英文作者：ZHAO Junjie;WANG Kaifang;CHEN Xusheng;MAO Zhonggui;Key Lab of Industrial Biotechnology,Ministry of Education,School of Biotechnology,Jiangnan University; 关键词：多肽 ; ε-聚赖氨酸 ; 基因组重排 ; 抗性筛选 ; 巴龙霉素 英文关键词：polypeptide;;ε-poly-L-lysine;;genome shuffling;;resistance screening;;paromomycin 中文刊名：ZYZZ 英文刊名：China Oils and Fats 机构：江南大学生物工程学院工业生物技术教育部重点实验室; 出版日期：2019-01-17 15:15 出版单位：中国油脂 年：2019 期：v.44;No.335 基金：江苏省产学研合作前瞻性联合研究项目(BY2016022-25) 语种：中文; 页：ZYZZ201901017 页数：6 CN：01 ISSN：61-1099/TS 分类号：81-86

摘要

聚赖氨酸产生菌Streptomyces albulus M-Z18的摇瓶产量较低,仅为1. 60 g/L。利用抗性筛选和基因组重排技术对S. albulus M-Z18进行菌种选育以获得高产ε-聚赖氨酸菌株。通过引入巴龙霉素抗性到S. albulus M-Z18中,获得两株性状优良的菌株S. albulus P-1和S. albulusP-2,ε-聚赖氨酸产量分别为2. 20 g/L和2. 16 g/L,单位菌体ε-聚赖氨酸合成能力分别为0. 41g/g和0. 39 g/g,作为基因组重排的亲本菌株;运用正交实验优化实验条件,最终获得一株高产ε-聚赖氨酸的融合子S. albulus G12,产量为2. 73 g/L,相比M-Z18提高了70. 63%。
        The ε-poly-L-lysine( ε-PL) producing strain Streptomyces albulus M-Z18 produced only 1. 60 g/L in shake flasks. S. albulus M-Z18 was bred by resistance screening and genome shuffling technology to get the ε-PL high-productivity strain. The paromomycin resistance was introduced into S. albulus M-Z18,and two strains with excellent traits,S. albulus P-1 and S. albulus P-2,were obtained with ε-PL production of 2. 20 g/L and 2. 16 g/L,respectively. The ε-PL biosynthetic capacity per strain was 0. 41 g/g and 0. 39 g/g,respectively. Using S. albulus P-1 and S. albulus P-2 as the parent strains for genome shuffling,the high-yield fusions were screened using protoplast fusion technology,and finally a fusion with high production of ε-PL was obtained. The ε-PL production of S. albulus G12 was 2. 73 g/L,an increase of 70. 63% compared with M-Z18.

引文

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