Prabha V, Gupta M, Seiffge D, Gupta K G
Department of Microbiology, Panjab University, Chandigarh, India.
Can J Microbiol. 1990 Feb;36(2):131-5. doi: 10.1139/m90-023.
Purification studies of 7 alpha-hydroxysteroid dehydrogenase (7 alpha-HSDH) (EC 1.1.1.159) from Escherichia coli 080 showed that 1.59-fold purification could be achieved by heating (60 degrees C for 10 min) the ultracentrifuged enzyme preparation, and 6.46-fold purification was achieved by subsequent precipitation with ammonium sulfate. Further purification on Sephadex G-100 gel gave 10.1-fold purification. After pooling and concentrating the active fractions obtained from the Sephadex G-100 filtration, an 11.1-fold purification was achieved using DEAE-cellulose chromatography. The purified enzyme produced a single band on polyacrylamide gel electrophoresis and its molecular weight was determined to be 54,000. The enzyme was immunogenic and showed immunoprecipitation with homologus antisera.
对来自大肠杆菌080的7α-羟基类固醇脱氢酶（7α-HSDH，EC 1.1.1.159）的纯化研究表明，通过对超速离心后的酶制剂进行加热（60℃，10分钟）可实现1.59倍的纯化，随后用硫酸铵沉淀可实现6.46倍的纯化。在Sephadex G - 100凝胶上进一步纯化得到10.1倍的纯化效果。合并并浓缩从Sephadex G - 100过滤获得的活性组分后，使用DEAE - 纤维素色谱法实现了11.1倍的纯化。纯化后的酶在聚丙烯酰胺凝胶电泳上呈现单一条带，其分子量测定为54,000。该酶具有免疫原性，与同源抗血清呈现免疫沉淀反应。
