农药残留快速检测固定化酶片的研究  

Study on Immobilized Enzyme for Rapid Detection of Pesticide Residues

作　　者：许娟[1] 李建科[1] 牛乐[1] 

机构地区：[1]陕西师范大学食品工程与营养科学学院,陕西西安710062

出　　处：《食品科学》2008年第6期268-272,共5页Food Science

基　　金：西安市科技攻关项目(GG05112);陕西省科技攻关项目(2004K01-G7)

摘　　要：采用硝酸纤维素膜(NC)和植物酯酶研究了农药残留快速检测固定化酶片。通过对酶用量、戊二醛浓度、牛血清白蛋白浓度、固定化pH值和温度的参数优化,建立了NC膜固定化植物酯酶的方法,即:20μl游离酶,0.05%戊二醛2μl,0.5%牛血清白蛋白15μl,pH7.0缓冲液体系,4℃下固定8h。对游离酶和固定化酶的特性进行了比较,发现固定化植物酯酶对底物的最适反应温度为35℃,比游离酶提高了5℃;最适pH为8.0,比游离酶提高了0.5个pH单位。游离酶和固定化酶对四种农药(敌敌畏、灭多威、氧乐果、久效磷)的敏感性进行了比较,发现固定化酶对四种农药敏感性都较游离酶有所提高,固定化酶更适合于农药的快速检测。Soybean phytoesterase （SPE） was used to immobilize on nitrocellulose membrane （NCM） in this study. Through the parameters optimization of enzyme dosage, glutaraldehyde （GA） concentration, bovine serum albumin （BSA） concentration, pH value and temperature, the method of NCM immobilizing SPE was established： adding 20 μl enzyme, 2 μl0.05% GA and 15μl0.5% BSA into test tube, then supperadding 0.03 mol/L pH 7.0 PBS buffer to 100μl, immobilizing for 8 h at 4 ℃. Comparing the characteristics between immobilized enzyme and free enzyme, the results showed that the optimum reaction temperature of immobilized enzyme is 35 ℃, 5 ℃ higher than that of free enzyme. The optimum pH value is 8.0, 0.5 higher than that of free enzyme; The sensitivity of immobilized enzyme to four pesticides （dichlorvos, methomyl, omethoate and monocrotophos） is better than that of free enzyme. So, it can be conclude that the immobilized enzyme is more suitable for rapid detection of pesticide.

关 键 词：植物酯酶 固定化 农残检测 

分 类 号：Q814[生物学—生物工程]