甲磺隆水解酶酶促反应体系的建立与特性研究  

Study on Reaction System and Characteristics of Metsulfuron-methyl Hydrolase

作　　者：黄星[1] 孙纪全[1] 潘继杰[1] 顾立锋[1] 孙笑非[1] 李顺鹏[1] 

机构地区：[1]南京农业大学生命科学学院农业部农业环境微生物工程重点开放实验室,江苏南京210095

出　　处：《农药学学报》2005年第4期329-333,共5页Chinese Journal of Pesticide Science

基　　金：国家自然科学基金(30500010);江苏省自然科学基金(BE2003343)等项目

摘　　要：在室内控制条件下对甲磺隆降解菌S113(Methylopila sp.)的水解酶进行了研究,发现该酶为胞内酶、非诱导酶.建立了甲磺隆水解酶的酶促反应体系:2 830μL Na2HPO4-NaH2PO4缓冲液(0.2 mol/L,pH 7.0),150μL甲磺隆(1000 mg/L),20μL粗酶液(约1.2μg粗蛋白),30℃水浴30 min.甲磺隆水解酶pH值稳定范围为6～9,最适pH值为8.0.水解酶热不稳定,45℃处理30 min,酶活下降59.22%,70℃处理30 min可完全灭活.测定了8种金属离子对水解酶的作用,发现1 mmol/L的Ca2+对酶活有促进作用.Metsulfuron-methyl hydrolase from metsulfuron-methyl degrading strain S113 （Methylopila sp. ） was studied. The hydrolase in the bacterium was endoemzyme. The enzymatic reaction system was found as follows： Na2HPO4-NaH2PO4 buffer 2 830 μL, metsulfuron-methyl （ 1 000 mg/L ） 150 μL, crude enzyme 20 μL （about 1.2 μg crude protein）, 30℃ water bathing 30 min. The characteristics of metsulfuron-methyl hydrolase was determined. The pH suitable for keeping enzyme was 6 - 9, with the optimum pH 8.0. The activity of enzyme was down to 59.22% and zero when treated at 45℃ for 30 rain and 70℃ for 30 min, respectively. Eight metal ions were chosen to study their effects on the metsulfuron-methyl hydrolase activity, and the result showed that 1 mmol/L Ca^2＋ can enhance the enzyme activity.

关 键 词：甲磺隆 水解酶 酶促反应体系 酶液特性 

分 类 号：Q556[生物学—生物化学]