头状链霉菌原生质体的制备及其诱变  

Preparation and Mutation of Protoplasts from Streptomyces caespitosus

作　　者：马文哲[1] 王鲁燕[1] 冯艳春 陈代杰[2] 许文思[3] 

机构地区：[1]中国药科大学生物制药学院,南京210009 [2]上海来益生物药物研究开发中心,201203 [3]上海医药工业研究院,200040

出　　处：《中国药科大学学报》2002年第1期62-65,共4页Journal of China Pharmaceutical University

摘　　要：目的 :建立头状链霉菌原生质体的制备方法 ,进而对其原生质体进行诱变 ,以期筛选到丝裂霉素 C的高产菌株。方法 :采用酶法制备原生质体 ,用紫外及激光进行原生质体诱变 ,HPLC检测突变株的生产能力。结果 :头状链霉菌原生质体制备的最佳条件为菌丝体培养基中含有 1 .5 %甘氨酸、2 5 %蔗糖 ,酶解液中含有溶菌酶5 mg/ ml、蜗牛酶 2 mg/ ml、纤维素酶 2 mg/ m,溶菌温度 33℃ ,酶解 2 h。对其原生质体进行诱变 ,激光诱变正突变率高于紫外诱变 ,经激光诱变后得到两株高产突变株 ,分别比出发菌株产量高 2 2 .2 %及 2 3.0 %。结论 :该方法有效地改善了此株头状链霉菌原生质体制备效果 ,用其进行激光诱变筛选到了丝裂霉素AIM The purpose is to develop the method of the preparation of protoplasts from Streptomyces caespitosus and to screen high-yielding mutant strains by mutation of the protoplasts. METHOD The protoplasts were prepared by enzyme and irradiated with UV and laser. The mutation result was determined by HPLC. RESULTS The optimum conditions for preparing the protoplasts from Streptomyces caespitosus were summarized as follows: to cultivate the mycelia in the medium containing 1.5%Gly and 25% sucrose and to lysis the mycelia in the solution containing 5 mg/ml lysozyme,2 mg/ml snailase and 2 mg/ml cellulase in the bath of 33℃, 2 h. The positive rate of mutants irradiated with laser was higher than that with UV and two high-yielding mutants were obtained by laser irradiation whose yield is 22.2% and 23.0% higher than that of the parent strain respectively. CONCLUSION The method greatly improved the preparation of protoplasts from Streptomyces caespitosus and high-yielding mutant strains could be obtained by laser irradiation of the protoplast.

关 键 词：头状链霉菌 原生质体 紫外诱变 激光诱变 制备 诱变 

分 类 号：Q939.9[生物学—微生物学]