酿酒酵母与糖化酵母原生质体的形成与再生

Research on Protoplast Formation and Regeneration of S.Cerevisiae and S.Diasticus

对影响酿酒酵母及糖化酵母形成与再生的因素——菌龄、酶浓度、温度、酶解时间、预处理剂的浓度和作用时间及渗透压稳定剂的选择做了初步的研究，确定了原生质体形成与再生的最佳条件：菌龄为13～15h，0．1％β-巯基乙醇作用10min；在30℃下，2．0％的蜗牛酶酿酒酵母、糖化酵母酶解1．0h；配制酶解液时渗透压稳定剂采用0．8mol／LKCl，而在稀释及配制再生培养基时则采用0．53mol／L蔗糖。

A series of single factor test and orthogonal test was used to determine the optimal conditions for protoplast formation and regeneration of S. cereusiae and S. diastaticus. The optimal conditions are that keeping yeast age on 13-15 hours, disposing it in 10 minutes by 0.1% β-Mercaptoethanol,then using 2.0% snaillase to disintegrate the cell wall of S. cereusiae and S. diastaticus for 1 hour in 30℃ warm water , using 0.8 mol/L KCl to confect snaillase solvent while using 0.53 mol/L sucrose to confect dilution and regenerational medium.