黑曲霉降酸菌株F1降解酒石酸关键酶的分离纯化

Isolation and Purification of Key Tartaric Acid Degrading Enzyme from Aspergillus niger F1

为得到黑曲霉菌株F1中具有降酸作用的蛋白,将黑曲霉菌株F1经酒石酸诱导发酵得到菌丝体,用液氮研磨破碎细胞壁,超声波辅助提取得到粗酶液,硫酸鱼精蛋白去除核酸,热变除杂蛋白,聚乙二醇浓缩,透析脱盐,DEAE-纤维素52阴离子交换层析,Sephadex G-75葡聚糖凝胶柱分子筛层析,最终得到该蛋白的单一组分,其纯化倍数为14.32,并确定该酶为脱氢酶类。

The purpose of this study was to prepare a key tartaric acid degrading enzyme from Aspergillus niger F1.Aspergillus niger F1 was cultured and the expression of tartaric acid degrading enzymes was induced by adding tartaric acid into the medium.Cultured mycelia of Aspergillus niger F1 were collected and homogenized in liquid nitrogen.Crude enzyme extract was obtained by ultrasonic assisted extraction,added with protamine sulfate for removing nucleic acids,denatured for removing unwanted proteins,concentrated by PEG treatment,dialyzed for desalting,and then purified by DEAE-cellulose 52 anion exchange chromatography and Sephadex G-75 gel filtration chromatography to obtain a single component with a purification factor of 14.32.The enzyme was proven to be a dehydrogenase.