原生质体诱变及融合选育那西肽高产菌株  

Screening of high nosiheptide-producing strain by protoplast mutation and fusion

作　　者：公维亮 薛正莲[1] 周扬[1] 张勇[1] 张奥[1] 沈强[1] 

机构地区：[1]安徽工程大学生物与化学工程学院,芜湖241000

出　　处：《中国抗生素杂志》2016年第5期335-339,381,共6页Chinese Journal of Antibiotics

基　　金：安徽省高校自然科学基金(No.KJ2013A048);国家大学生创新创业训练计划(No.201410363025)

摘　　要：目的提高工业发酵那西肽生产水平,获得那西肽高产菌株。方法利用溶菌酶剥离菌体细胞壁制备原生质体,通过等离子(ARTP)诱变与双亲灭活原生质体融合技术选育那西肽高产菌株。结果通过对原生质体制备参数甘氨酸浓度,溶菌酶浓度以及酶解时间优化,确定了甘氨酸最适浓度0.6%,溶菌酶最适浓度50mg/m L,酶解时间70min。通过ARTP诱变筛选出突变株AH-12产量提高18%。亲本最佳紫外灭活时间为4min,最佳热灭活条件为65℃水浴50min。对融合子进行摇瓶选育,得到了GSM-4,GSM-5两株效价提高15%以上菌株。结论利用原生质体诱变及融合方法可获得那西肽高产菌株。Objective To raise the level of nosiheptide fermentation, and obtain the mutants with improved nosiheptide production. Methods Screening high production strain of nosiheptide by ARTP mutagenesis and fusion of inactivated parental protoplasts prepared by using lysozyme. Results The optimum conditions of preparing protoplast were as follows： the mycelia of Streptomyces actuosu were processed with 50mg/mL lysozyme for 70min and the seed medium containing 0.6% glycine. The strain of AH-12 was obtained by ARTP mutagenesis and the increased yield was about 18% higher than that of the parent strain. Before fusion, one of the strain of parent is inactivated by ultraviolet for 4 minutes and the other one in the conditions of 65℃ water for 50 minutes. Two high yielding strains of GSM-4 and GSM-5 were isolated and the increased yield was about 15% higher than that of the parent strain. Conclusion It is a way to screen high nosiheptide-producing strain by protoplast mutation and fusion.

关 键 词：那西肽 原生质体制备 原生质体诱变 原生质体融合 

分 类 号：R978.1[医药卫生—药品]