双亲灭活制备粘质沙雷氏菌和红曲霉的跨界产色素融合子

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。

Preparation of Cross-border Integration of Sub-pigment in SerratiaMarcescens and Monascus by Parents Inactivated Protoplast Technology

Objective: To prepare inter-kingdom pigmented fusant from Serratia marcescens and Monascus by double parentsinactivated protoplasts method and determine the inhibition activity of the pigmented fusants. Methods:The protoplast of Serratiamarcescens was obtained by 0.2% lysozyme treatment and then inactivated by heat treatment, while the protoplast of Monascus wasobtained by enzyme mixture with 0.8% lysozyme, 1.2% snail enzyme and 1.6% cellulase, then inactivated by ultraoviolet treatment. Theprotoplast fusion of double inactivated parents was carried out using fusion solution with 25% polyethylene glycol. The morphology andpigment produced ability of the fusants were observed, while the inhibition activity of the pigment extrative on Staphylococcus aureu wastested. Results: Under the optimal conditions, the protoplast formation rate of Serratia marcescens and Monascus was 92.58% and 106/mL, respectively. The protoplast inactivated rate of both microbes was 100%. Thirteen protoplast fusants was prepared while nine ofthem can produce pigment with 1×10-5% fusion rate. Pigment extractive with 95% ethanol from eight protoplast fusants showedinhibiton on Staphylococcus aureus of varing degree. Conclusion:Inter-kingdom protoplast fusants of Serratia marcescens and Monascuswith antimicrobial activity can be prepared by double parents inactivated protoplasts method.