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目的:采用双亲灭活原生质体技术制备粘质沙雷氏菌和红曲霉的跨界产色素融合子,并测定其抑菌活性。方法:经0.2%溶菌酶处理获得粘质沙雷氏菌的原生质体并热灭活;经混合酶(0.8%溶菌酶+1.2%蜗牛酶+1.6%纤维素酶)处理获得红曲霉的原生质体并紫外灭活;用含25%PEG的原生质体融合剂进行促融合与再生。观察融合子的菌落形态和色素合成能力,测定融合子色素提取物对金黄色葡萄球菌的抑制活性。结果:在优化条件下,粘质沙雷氏菌原生质体的形成率为92.58%,红曲霉原生质体形成数约为106个/mL,两菌原生质体灭活率均为100%。共获得13个融合子,9个能产红色素,融合率为1×10-5%。其中8个融合子的95%乙醇提取物对金黄色葡萄球菌表现出不同程度的抑制。结论:采用双亲灭活原生质体技术,能够制备具有抑菌活性的粘质沙雷氏菌和红曲霉的跨界产色素融合子。
OBJECTIVE: To prepare cross-border pigment fusion of Serratia marcescens and Monascus spp. By in-amphiphilic inactivation protoplast technology and determine its antibacterial activity. Methods: Protoplasts of Serratia marcescens were obtained by 0.2% lysozyme treatment and protoplasts of Monascus spp. Were isolated by a mixed enzyme (0.8% lysozyme + 1.2% snail enzyme + 1.6% cellulase) And inactivated by UV light. The fusion and regeneration were performed with 25% PEG protoplast fusion. The colony morphology and biosynthesis ability of the fusion were observed, and the inhibitory activity of the fusion pigment on Staphylococcus aureus was determined. Results: Under the optimal conditions, the formation rate of Serratia marcescens protoplasts was 92.58%, the number of protoplasts of Monascus was about 106 / mL, and the inactivation rates of both protoplasts were both 100%. A total of 13 fusants were obtained, 9 of them could produce red pigment, and the fusion rate was 1 × 10-5%. Among them, 95% ethanol extract of eight fusants showed different degrees of inhibition on Staphylococcus aureus. Conclusion: Cross-bifunctional pigment fusion of Serratia marcescens and Monascus spp. With antibacterial activity can be prepared by in-amparent inactivated protoplast technology.