原生质体再生与诱变在硫霉素产生菌选育中的应用  

Improvement of thienamycin producing strain by regeneration and mutagenesis of protoplasts

作　　者：崔大鹏[1,2] 石莲英 

机构地区：[1]中国医学科学院 [2]中国协和医科大学医药生物技术研究所

出　　处：《中国抗生素杂志》1999年第4期265-268,共4页Chinese Journal of Antibiotics

基　　金：国家"九五"攻关项目

摘　　要：以硫霉素产生菌卡特利链霉菌３５８（Ｓｔｒｅｐｔｏｍｙｃｅｓｃａｔｌｅｙａ３５８）为原始菌株，用原生质体形成与再生结合物理、化学诱变的方法进行菌种选育。在最适条件下原生质体释放浓度和再生率分别为１．６×１０９／ｍｌ和２９．５％。原生质体再生处理后，硫霉素产量变异向正方向移动，筛选到１株ＰＲ－１１７，相对效价提高６０％。分别以菌株ＰＲ－１１７的孢子和原生质体进行紫外线及亚硫酸氢钠诱变，原生质体对诱变剂的敏感性强于孢子，致死率增加，效价分布更分散。原生质体与孢子紫外线诱变正变率分别为１６．５％和９％，亚硫酸氢钠诱变的正变率都为４％。从原生质体紫外线诱变株中筛选到１株ＰＲｕ－３３６，相对效价比原始菌株提高９０％。Protoplast regeneration combined with physical and chemical mutagenization was performed to improve thienamycin production of Streptomyces cattleya . Under the optimum condition, the concentration and regeneration frequency of protoplast were 1.6×10 9/ml and 29.5% respectively. After protoplast regeneration, the curve of normal distribution of relative titer had a positive mutation and mutant PR 117 was obtained, whose relative titer was 60 percent higher than original strains. Spores and protoplasts of strain PR 117 were treated with UV and NaHSO 3 respectively. Results showed that compared with spores, protoplasts were more sensitive to mutagen, death rate increased and the curve of distribution of relative titer was wider after these treatments. When treated with UV irradiation, the rate of positive mutants of protoplasts and spores were 16.5% and 9% respectively and mutant PRu 336 was obtained, whose relative titer was 90% higher than that of original strain. When treated with NaHSO 3, this positive mutation rate of protoplasts and spores were both 4%.

关 键 词：硫霉素 卡特利链霉菌 原生质体再生 

分 类 号：Q939.13[生物学—微生物学] TQ465.1[化学工程—制药化工]