常温常压等离子体诱变选育高产L-异亮氨酸谷氨酸棒杆菌  

Mutation breeding of high-yield L-isoleucine Corynebacterium glutamicum by atmospheric and room temperature plasmas mutagenesis

作　　者：孔帅 陈敏[1] 郑美娟[1] 许鹏飞 吕育财[1] 谢飞 周宜平 龚大春[1] KONG Shuai;CHEN Min;ZHENG Meijuan;XU Pengfei;LV Yucai;XIE Fei;ZHOU Yiping;GONG Dachun(Hubei Engineering Technology Research Center for Biological Jiaosu,China Three Gorges University,Yichang 443002,China;Yichang Sanxia Pharmaceutical Co.,Ltd.,Yichang 443002,China)

机构地区：[1]三峡大学湖北省生物酵素工程技术研究中心,湖北宜昌443002 [2]宜昌三峡制药有限公司,湖北宜昌443002

出　　处：《中国酿造》2019年第7期76-79,共4页China Brewing

基　　金：国家自然基金项目(21776162);宜昌市科技攻关项目(A17-302-a04)

摘　　要：以谷氨酸棒杆菌(Corynebacterium glutamicum)23798为原始菌株,对其进行常温常压等离子体(ARTP)诱变,以磺胺胍抗性和氨基酸与茚三酮特异显色为筛选标记,以期得到高产L-异亮氨酸的诱变谷氨酸棒杆菌,并对其遗传稳定性进行研究。结果表明,原始菌株23798经过ARTP诱变处理180s后,经0.4mg/mL磺胺胍抗性筛选、多孔板高通量筛选、发酵培养复筛,选育出一株高产L-异亮氨酸诱变谷氨酸棒杆菌(Corynebacterium glutamicum)B1。该菌株在摇瓶中发酵培养48h,L-异亮氨酸产量达18.5g/L,比原始菌株提高62.03%,且遗传性状稳定。Using Corynebacterium glutamicum 23798 as original strain,the original strain was mutated by atmospheric and room temperature plasma (ARTP). Using the sulfaguanidine resistance and the characteristic color of amino acid and ninhydrine as selection markers,the high-yield L-isoleucine C. glutamicum mutant was obtained,and its genetic stability was studied. The results showed that after the original strain 23798 was mutated for 180 s by ARTP,a high-yield L-isoleucine C. glutamicum mutant strain B1 was bred by 0.4 mg/ml sulfaguanidine resistance screening,perforated plate high-throughput screening and fermentation rescreening. The fermentation of strain with stable genetic character was carried out in shake flask for 48 h,and the L-leucine yield reached 18.5 g/L,which was 62.03% higher than that of the original strain.

关 键 词：谷氨酸棒杆菌 常温常压等离子体 磺胺胍抗性标记 高通量筛选 L-异亮氨酸 

分 类 号：TQ922.9[轻工技术与工程—发酵工程]