作为单加氧酶的细胞色素P450 BM-3的纯化

Purification of cytochrome P450 BM-3 as a monooxygenase.

作者信息

Jun Huang, Lehe Mei, Qing Sheng, Dongqiang Lin, Shanjing Yao

机构信息

Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou, PR China.

出版信息

Protein Pept Lett. 2005 May;12(4):327-31. doi: 10.2174/0929866053765653.

DOI:10.2174/0929866053765653

PMID:15907176

Abstract

After investigating two anion-exchange resins, the purification factor and activity yields of P450 BM-3 were higher with Resource Q than with DEAE-Sepharose FF. Screening of HIC media showed that Source 15ISO was the most suitable for purification of P450 BM-3. An effective isolation and purification procedure of P450 BM-3 was developed and included three steps: 35%-70% saturation (NH(4))(2)SO(4) precipitation, Source 15ISO hydrophobic interaction chromatograph and Sephacryl S-200 gel filtration chromatography. Using this protocol, the purification factor and P450 BM-3 activity recovery was 13.5 and 13.7%, respectively.

摘要

在研究了两种阴离子交换树脂后，Resource Q对P450 BM-3的纯化因子和活性回收率高于DEAE-Sepharose FF。疏水相互作用色谱介质筛选表明，Source 15ISO最适合纯化P450 BM-3。开发了一种有效的P450 BM-3分离纯化方法，包括三步：35%-70%饱和度的硫酸铵沉淀、Source 15ISO疏水相互作用色谱和Sephacryl S-200凝胶过滤色谱。使用该方法，纯化因子和P450 BM-3活性回收率分别为13.5和13.7%。