固定化植物酯酶的制备及应用  

Preparation and application of immobilized plant-esterase

作　　者：范钦华[1] 周小凡[2] 

机构地区：[1]南京林业大学生物与环境学院,江苏南京210037 [2]南京林业大学轻工科学与工程学院,江苏南京210037

出　　处：《现代化工》2015年第3期112-115,117,共5页Modern Chemical Industry

基　　金：南京林业大学创新项目(163030045);江苏高校优势学科建设工程资助项目

摘　　要：以麦草秸秆为原料,经环氧氯丙烷和乙二胺改性,戊二醛交联,制备了植物酯酶的固定化材料,研究了固定化植物酯酶的最优条件和固定化酶的酶学性质.结果表明,加酶量为20 mL/g（改性麦杆）,固定化时间为6h,温度为35℃,pH为7.0条件下,固定化效果最好,酶活回收率可达46％.固定化酶的pH稳定性、热稳定性和贮存稳定性都明显优于游离酶.固定化酶的米氏常数为41 mmol/L,固定化酶与底物的亲合力低于游离酶.建立了酶抑制反应的标准曲线和检测敌敌畏的方法,线性范围为1.0×10^-4～6.25×10^-4mg/L,检测下限为0.06μg/L.The wheat straw is modified by using epichlorohydrin, ethanediamine and glutaraldehyde to prepare the immobilized material. The optimum immobilization conditions and characterizations of the immobilized esterase are investigated. The results show that the good immobilization effect can be achieved under the following conditions ：20 mL/ g of crude enzyme （with respect to modified wheat straw）, 6 hours of immobilized time, 35℃ of immobilization temperature and 7.0 of pH. The recovery rate of the enzyme activity is 46%. The pH, thermal and storage stability of the immobilized enzyme are greatly improved. The Michaelis constant of the immobilized esterase is 41 mmol/L. The standard curve and method to determination of dichlorvos by enzyme inhibition are established. The linear ranges for the determination of dichlorvos are 1.0×10^-4～6.25×10^-4mg/L. The detection limit is 0. 06 μg/L.

关 键 词：麦草秸秆 固定化 植物酯酶 敌敌畏 

分 类 号：TS201.2[轻工技术与工程—食品科学]