杀灭菊酯降解菌菌株的分离鉴定和降解纯化酶的特性研究  

Isolation and Identification of Fenvalerate Degrading Bacteria and Characteristics of Purified Degrading Enzymes

作　　者：阎华[1] 于博[1] YAN Hua;YU Bo(College of Food Science and Technology,Hubei University of Arts and Sciences,Xiangyang 441053,China)

机构地区：[1]湖北文理学院食品科学技术学院,湖北襄阳441053

出　　处：《东北农业科学》2023年第4期56-61,94,共7页Journal of Northeast Agricultural Sciences

基　　金：湖北省高等学校优秀中青年科技创新团队计划项目(T201616);湖北文理学院食品新型工业化学科群建设项目(XKQ08321)。

摘　　要：从湖北省襄阳市南漳王冠茶园表层土分离得到降解杀灭菊酯农药的菌株TK-2。菌株TK-2革兰氏染色为阴性,镜检为长棒状,生理生化特征和16S rDNA序列特征符合铜绿假单胞菌典型特征,综合鉴定其为铜绿假单胞菌。TK-2在添加杀灭菊酯的无机盐培养基培养72 h后,获得最高产量的杀灭菊酯降解酶,酶活力为10.8 U/mL。TK-2杀灭菊酯降解酶经过DEAE-52和CM-52纯化后,纯化倍数达到3.4倍,酶活力回收率达到56.2%,凝胶电泳后分析得到酶的分子量为83.6 kDa。TK-2杀灭菊酯降解纯化酶酶促反应的最适温度是35℃,25~45℃稳定性良好;酶促反应的最适pH值是8.0,pH 7.0~9.0稳定性良好,对底物反应的米氏常数Km是0.738 mmol/L,最大反应速率Vmax是1.123 mmol/(L·min)。Strain TK-2 was isolated from the topsoil of Nanzhang Wangguan Tea Garden in Xiangyang City, Hubei Province. The strain TK-2 was Gram-negative and long rod-shaped by microscopic examination. Its physiological and biochemical characteristics, as well as the 16S rDNA sequence characteristics, corresponded to the typical features of Pseudomonas aeruginosa. P. aeruginosa TK-2 was cultured in inorganic salt medium supplemented with fenvalerate for 72 h. The highest yield of fenvalerate degrading enzyme was obtained, with an enzyme activity of 10.8U/mL. After being purified by DEAE-52 and CM-52, the fenvalerate degrading enzyme of P. aeruginosa TK-2reached 3.4 times, and the recovery rate of enzyme activity reached 56.2%. The molecular weight of the enzyme was83.6 kDa after gel electrophoresis. The optimum temperature for degradation and purification of fenvalerate by P. aeruginosa TK-2 was 35℃, exhibiting good stability within the range of 25-45℃. The optimal pH value for the enzymatic reaction was 8.0, with good stability in the pH range of 7.0-9.0. The Michaelis constant(Km) for the substrate reaction was 0.738 mmol/L, and the maximum reaction rate Vmax was 1.123 mmol/(L·min).

关 键 词：铜绿假单胞菌 杀灭菊酯 纯化酶 降解特性 

分 类 号：X172[环境科学与工程—环境科学]