摘要
目的研究井冈霉素产生菌吸水链霉菌井冈变种的原生质体制备与再生的最佳条件。方法使用溶菌酶水解菌体细胞壁法,分别考察影响吸水链霉菌井冈变种原生质体的制备与再生的各种因素。结果在R2YE液体培养基中,一级菌丝体的最佳培养时间为24 h,二级菌丝体的最佳培养时间为16 h,最佳甘氨酸质量浓度为5 g.L-1,最佳溶菌酶质量浓度为2 g.L-1,最佳酶解时间为60 min,最佳再生培养基为R2YE,原生质体的再生率最高可达到15.79%。结论本实验确定了吸水链霉菌井冈变种的原生质体制备与再生的最佳条件,能够得到较高的再生率。
Objective To obtain the optimal condition of protoplasts formation and the ability of regeneration of Streptomyces hygrascopicus var. Jinggangensis Yen. Methods All factors about protoplasts formation and regeneration of S. hygrascopicus were studied by the method of losozyme hydrolyzation cell wall. Result and conclusions The experiments results showed the protoplast regeneration rate could approach 15.79% under the follow conditions： primary mycelia were cultivated for 24 h in R2YE liquid medium, and then transferred to the same medium with 5 g·L^-1 glycine for the secondary mycelia cultivation. After 16 h cultivation, the mycelia were collected, washed and lysed by lysozyme at the optimal concentration of 2 g·L^-1 for 60 min, Then spread the prepared protoplast on R2YE solid medium for regeneration.
出处
《沈阳药科大学学报》
CAS
CSCD
北大核心
2007年第5期315-319,324,共6页
Journal of Shenyang Pharmaceutical University
关键词
吸水链霉菌
原生质体制备
原生质体再生
Streptomyces hygrascopicus
protoplast formation
protoplast regeneration
