真菌胞内蛋白质提取方法的建立

目的：研究不同的提取方法对白假丝酵母和茄病镰刀菌胞内蛋白质浓度、种类及数量的影响，为建立一种稳定可靠的真菌胞内蛋白质提取方法提供依据。方法：分别以白假丝酵母菌和茄病镰刀菌为研究对象，培养时间为36、48和72 h，超声波工作时间为20 s、3 min、5 min，10 min，超声波功率为332.5和475.0 W，通过蛋白质浓度检测和SDS-PAGE蛋白质电泳分析，比较不同条件下提取的蛋白质浓度、种类及数量，观察不同因素对蛋白质提取效果的影响。结果：2株真菌均在培养36 h时提取胞内蛋白质的浓度最大，白假丝酵母在功率332.5 W、超声3 min时得到胞内蛋白质的数量和种类最多，茄病镰刀菌在功率332.5 W、超声10 min时得到蛋白质的数量和种类最多。结论：使用超声波法与玻璃珠研磨法结合提取胞内蛋白质，从菌株的培养时间、超声波工作时间和功率3方面建立了真菌胞内蛋白质的最佳提取方法。

Establishment of intraceilular protein extraction methods from fungus

Objective To study the effect of different cultivate time,ultrasonic time and ultrasonic power on the concentration,type and quantity of the intracellular protein from Candida albicans and Fusarium solina and to provide the theoretical basis for establishing a reliable and stable extraction method for intracellular protein from fungus.Methods Candida albicans and Fusarium solina were cultured.The different parameters included the culture time(36,48,and 72 h),ultrasonic time(20 s,3 min,5 min,and 10 min) and ultrasonic power(332.5 W and 475.0 W).The concentrations,species and quantities of protein under different factors were detected by the protein concentration detection and SDS-PAGE electrophoresis analysis.Results Both strains got the most quantity of protein when cultivated for 36 h.The most quantity and species of Candida albicans were obtained when the ultrasonic power was 332.5 W and ultrasonic time was 3 min,but as to Fusarium solina,the corresponding parameters were 332.5 W and 10 min,respectively.Conclusion The intracellular protein is extracted by compositely using ultrasonic method and glass bead transformation method.The optimal extraction method of intracellular protein from fungus is developed from the aspects of culture time,ultrasonic power and ultrasonic working time of strains.