污白干酪菌核糖核酸酶的分离纯化和理化性质

Purification and Characterization of Ribonuclease from Tyromyces amygdalinus

采用DEAE-cellulose阴离子交换层析、CM-cellulose阳离子交换层析和FPLC-Superdex-75凝胶过滤层析,从污白干酪菌(Tyromyces amygdalinus)干子实体纯化得到一种核糖核酸酶,SDS-PAGE电泳检测其为单一蛋白,分子量为25 k Da。最适反应条件为60℃,p H4.4。该酶对几种寡聚核糖核苷酸的水解活性低于对t-RNA的水解活性。

A novel ribonuclease （RNase） was purified from dry fruit bodies of Tyromyces amygdalinus using a purification procedure which involved ion exchange chromatography on DEAE-cellulose, CM-cellulose and gel filteration chromatography by FPLC on superdex 75HR 10/300 column. Combine with the result from FPLC and SDS-PAGE, it was estimated that this ri- bonuclease was a monomeric protein with a molecular weight of 25 kD. The optimum temperature of this RNase is 60℃, and optimum pH of it is 4.4. The activity of this enzyme toward poly A and poly C was lower than that of t-RNA.