Arthrobacter globiformis酯酶基因的克隆、表达和酶活性测定
Cloning and Expression of the Esterase Gene of Arthrobacter globiformis and the Activity Assay of the Esterase
-
摘要: 采用聚合酶链反应(PCR)的方法扩增了A rthrobacter g lobiform is中酯酶编码基因,与载体质粒连接后在大肠杆菌BL 21(DE 3)中进行表达。以包涵体形式表达的蛋白在8 m o l/L尿素作用下溶解,经过透析复性后获得了具有活性的粗蛋白。产物活性实验表明,酯酶表现出较高的催化活性,为菊酯作为杀虫剂的应用奠定了基础。Abstract: A novel esterase which stereoselectively hydrolyzes the( )-trans stereoisomer of ethyl chrysanthemate to produce( )-trans-chrysanthemic acid was discussed.The gene coding for the esterase was amplified from Arthrobacter globiformis by polymerase chain reaction(PCR).The gene was inserted into a vector and expressed in Escherichia coli BL21(DE3).The protein expressed in body was dissolve in 8 mol/L urea before dialysis.The purified crude enzyme show high catalysis activity.It may suggest the possibility that( )-trans-chrysanthemate is identified as the insecticide.