应用毛细管气相色谱法检测棉铃虫细胞色素P450 O-脱甲基活性

Using capillary gas chromatography detect the activity of O-demethylation of cytochrome P450 in Helicoverpa armigera

依据毛细管气相色谱法灵敏度高 ,样品用量少 ,分辨率高的特点 ,建立了一个用于检测棉铃虫Helicoverpaarmigera幼虫中肠和脂肪体细胞色素P45 0O -脱甲基活性产物的小型反应体系 ( 875 μL) ,其中含 0 1mol L ,pH 7 8Tris HCl缓冲液 ,0 5 μmol,NADPH ,7 5 μmolMgCl2 ,0 0 85 %BSA ,0 1 5 7mmol对硝基苯甲醚及酶液。利用毛细管气相色谱直接检测该反应体系中棉铃虫细胞色素P45 0的O -脱甲基产物对硝基苯酚。

A 875μL of reaction system determining the activity of p\| nitroanisole O\|demethylation by cytochrome p450 was developed based on the traits of the capillary gas chromatography(CGC), which has higher sensitivity, less sample volume and higher resolution power than spectra photometry. The reaction system contained 0\^1 mol/L, pH 7\^8 Tris\|HCl buffer, 0\^5 μmol NADPH, 7\^5 μmol MgCl 2, 0\^085% BSA,0\^157 mmol p nitroanisole and enzyme. The p\| nitrophenol,the product of p\| nitroanisole O\|demethylation by cytochrome P450 from the midgut and fat body of H.armigera larva was detected by CGC.