马克斯克鲁维酵母原生质体制备和再生条件的研究

Study on the Formation & Regeneration of K.Marxianus Protoplast

研究了菌龄、酶解温度、酶解时间、酶浓度4个单因素对马克斯克鲁维酵母原生质体形成率和再生率的影响,采用菌龄、酶解温度、酶解时间、酶浓度、稳渗剂进行5因素4水平的正交试验,分析各因素的极差值,评价各因素的影响程度。结果表明,酶浓度的极差值最高,其次为酶解时间,然后依次为菌龄、稳渗剂、酶解温度,酶浓度为影响原生质体形成的主要因素,原生质体形成和再生的最佳条件为:菌龄为12h,蜗牛酶浓度为1.5%,酶解时间为1h,酶解温度为28℃,稳渗剂为KCl,在此条件下,原生质体形成率为92.3%,再生率为29.6%。

The effects of four single factors including hypha age, enzymolysis temperature, enzymolysis time and enzyme concentration on the formation rate and regeneration rate of K.Marxianus protoplast were studied. Five-factor and four- level orthogonal experiments were designed to study the range value and to evaluate the influencing effects of each single factor, The results indicated that the range value sequence from the highest to the lowest were enzyme concentration, enzymolysis time, hypha age, osmotant and enzymolysis temperature. Enzyme concentration was the main influencing factor for the formation of protoplast. And the optimum conditions for K.Marxianus protoplast generation and regeneration were summed up as follows： hypha age was 12 h, snail enzyme concentration was 1.5 %, enzymatic hydrolysis time was 1 h, enzymatic temperature was at 28 ℃ and potassium chloride was selected as osmotants. Under such conditions, the formation rate and regeneration rate of protoplast were 92.3 % and 29.6 % respectively.