【摘要】 从高效降解真菌镰孢霉属WZ-Ⅰ(FusariumLK.ex Fx)中提取了降解酶,研究了该降解酶的分离条件及对毒死蜱的降解特性,研究表明,其胞内酶对毒死蜱的降解率高达60.8%,细胞碎片对毒死蜱的降解率为48%,但由(NH4)2SO4沉淀提取的胞外酶液对毒死蜱的降解率仅为11.3%,经8次非诱导条件下培养提取粗酶液,酶活力损失较少,判断WZ-Ⅰ菌株的毒死蜱降解酶为胞内酶且属于组成酶.以牛血清白蛋白为标准蛋白测得粗提酶中可溶性蛋白的含量为3.36mg.mL-1;该酶对毒死蜱的酶促降解最适pH为6.8,在pH 6.0~9.0之间都有较高的活性;最适温度为40℃,在实验温度范围(20~50℃)内该降解酶均具有较好的降解活性,但在55℃时,酶活迅速降低,降低到最高酶活力的41%.测得粗提酶中其米氏常数Km为1.049 26mmol.L-1,vmax为0.253 5μmol.(mg.min)-1.研究结果表明该酶具有较好的热稳定性和pH稳定性,对热和pH均具有较好的耐受力,高效降解真菌WZ-Ⅰ所产生的胞内酶对毒死蜱具有较好的降解效果.更多还原

【Abstract】 Degradation characteristics of chlorpyrifos insecticides was determined by the crude enzyme extracted from the isolated strain WZ-Ⅰ（Fusarium LK.ex Fx）.The best separating condition and the degrading characteristic of chlorpyrifos were studied. Rate of degradation for chlorpyrifos by its intracellular enzyme,extracellular enzyme and cell fragment was 60.8%,11.3% and 48%,respectively.The degrading enzyme was extracted after this fungus was incubated for 8 generations in the condition of non-inducement,and its enzymic activity lost less,the results show that this enzyme is an intracellular and connatural enzyme.The solubility protein of the crude enzyme was determined with Albumin（bovine serum） as standard protein and the solubility protein of the crude enzyme was 3.36 mg·mL^-1. The pH optimum for crude enzyme was 6.8 for enzymatic degradation of chlorpyrifos,and it had comparatively high activity in the range of pH 6.0～9.0.The optimum temperature for enzymatic activity was at 40℃,it still had comparatively high activity in the range of temperature 20～50℃,the activity of enzyme rapidly reduced at 55℃,its activity was 41% of the maximal activity.The crude enzyme showed K_m value for chlorpyrifos of（1.049 26）mmol·L^-1,and the maximal enzymatic degradation rate was（0.253 5）μmol·（mg·min）^-1.Additional experimental evidence suggests that the enzyme had the stability of endure for temperature and pH,the crude enzyme of fungus WZ-Ⅰcould effectively degrade chlorpyrifos.更多还原