【摘要】 以青霉素(Penicillin)和链霉素(Streptomycin)分别作为克东腐乳发酵生产优势菌株藤黄微球菌(Micrococcus luteus)QSY和QSG的抗药性标记,根据原生质体形成率和再生率优化溶菌酶浓度、酶解时间、酶解温度和酶解pH值。实验结果表明:40μg/mL的青霉素和140μg/mL的链霉素可作为2株菌的抗性标记。对数生长末期的QSG原生质体最佳形成和再生的条件为:溶菌酶浓度3mg/mL、酶解时间60min、酶解温度37℃和酶解体系pH6.5;对数生长末期的QSY原生质体最佳形成和再生的条件为:溶菌酶浓度2mg/mL、酶解时间30min、酶解温度34℃和酶解体系pH6.5。更多还原

【Abstract】 Penicillin and Streptomycin were chosen as the sign of resistance to antibiotics of QSG snd QSY respectively and concentration of Penicillin of 40 μg/mL and Streptomycin of 140 μg/mL could be used for detecting of fusion cell. The effects of culture time, lysozyme concentration, enzymolysis time, enzymolysis temperature and enzymolysis pH on protoplast formation and regeneration of strains QSG and QSY were studied. The results showed that the best conditions of QSG protoplast formation and regeneration were lysozyme concentration of 3 mg/mL, enzymolysis time of 60 min, enzymolysis temperature of 37 ℃ , enzymolysis pH6.5, and the protoplast formation rates and regeneration rates were 84.2% and 25.4% respectively. The best conditions of QSY protoplast formation and regeneration were lysozyme concentration of 2 mg/mL, enzymolysis time of 30 min, enzymolysis temperature of 34 ℃, enzymolysis pH6.5, and the protoplast formation rates and regeneration rates were 93.3% and 38.4% respectively. These results and data would provide important parameters for the improvement of strains QSG and QSY through the protoplasttechnology.更多还原