采用硫酸铵分级分离、Sephadex G-100凝胶过滤,DEAE-cellulose 离子交换层析以及5′-AMP-Sepharose 4B 亲和层析,从猪肝微粒体中纯化得到可溶性的 NADH-细胞色素 b_5还原酶,提纯倍数为750～800,总回收率为40％左右。纯化的酶呈典型的黄素蛋白吸收光谱,最大吸收波长273,390和460nm,在415和475nm 处有一肩峰,A273/A460比值为5.8。荧光光谱分析该纯酶含 FAD 基团。在 SDS—聚丙烯酰胺凝胶电泳板上呈单一的蛋白质区带。分子 Purification from swine liver microsomes resulted in soluble NADH cytochrome b_5 reduction using ammonium sulfate fractionation, Sephadex G-100 gel filtration, DEAE-cellulose ion exchange chromatography, and 5’-AMP-Sepharose 4B affinity chromatography Enzyme, purification factor is 750 ~ 800, the total recovery is about 40%. The purified enzyme exhibits a typical flavoprotein absorption spectrum with maximum absorption wavelengths of 273, 390 and 460 nm, a shoulder at 415 and 475 nm, and A273 / A460 ratio of 5.8. Fluorescence spectroscopy analysis of the pure enzyme containing FAD group. On SDS-polyacrylamide gel electrophoresis showed a single protein band. molecular