贝莱斯芽孢杆菌(Bacillus velezensis)CF57中烟嘧磺隆降解酶的分离与鉴定  

Isolation and identification of nicosulfuron-degrading enzymes from Bacillus velezensis CF57

作　　者：张晨芳 司贺龙[1] 陈来 梁小菊 张金林[1] ZHANG Chenfang;SI Helong;CHEN Lai;LIANG Xiaoju;ZHANG Jinlin(College of Plant Protection,Hebei Agricultural University,Baoding 071001,China;Agricultural Technology Extension Station of Fengnan Town,Fengnan district,Tangshan city,Tangshan 063300,China)

机构地区：[1]河北农业大学植物保护学院,河北保定071001 [2]河北省唐山市丰南区丰南镇农业技术推广站,河北唐山063300

出　　处：《河北农业大学学报》2020年第4期43-48,共6页Journal of Hebei Agricultural University

基　　金：河北省现代农业产业技术体系玉产业创新团队(HBCT2018020205);国家自然科学基金(31471786).

摘　　要：为明确贝莱斯芽孢杆菌(Bacillus velezensis)CF57参与降解烟嘧磺隆的关键降解酶,本试验通过降解酶定域试验确定了贝莱斯芽孢杆菌(B.velezensis)CF57中对烟嘧磺隆具有降解作用的酶主要为胞外酶。在此基础上,采用丙酮沉淀法、DEAE-FF阴离子交换层析柱和PAGE法对胞外酶进行分离纯化,并结合水解圈法和高效液相色谱(HPLC)对分离得到的活性组分进行检测,选择其中活性较高的3种降解酶(P3-4、P4-2和P4-4)进行质谱测定。经序列比对,得知这3种降解酶分别为糖磷酸异构酶(P3-4)、亮氨酸氨基肽酶(P4-2)和精氨酸酶(P4-4),其对烟嘧磺隆的酶比活分别为0.0418、0.0368和0.0382μmoL/(min·mg)。该研究结果为进一步深入研究降解酶的酶学特性及代谢机理奠定基础。To determine the key degrading enzymes of Bacillus velezensis CF57 involved in the degradation of nicosulfuron,the extracellular enzyme was determined to be the key site for the degradation of nicosulfuron by Bacillus velezensis CF57 by means of enzyme localization test.Then,on this basis,the extracellular enzymes were separated and purified by acetone precipitation method,DEAE-FF anion exchange chromatography column and PAGE method,and the active components were detected by hydrolytic circle method and HPLC.Three kinds of degrading enzymes(P3-4,P4-2,P4-4)with high activity were selected for MALDI–TOF–MS.The sequence comparison showed that the three degrading enzymes were sugar phosphate isomerase(P3-4),probable cytosol aminopeptidase(P4-2)and arginase(P4-4),respectively.Their specific enzyme activity to nicosulfuron was 0.0418,0.0368 and 0.0382μmoL/(min·mg).These results lay a foundation for further study on the enzymatic characteristics and metabolic mechanism of degrading enzymes.

关 键 词：烟嘧磺隆 贝莱斯芽孢杆菌 酶 降解 

分 类 号：S482.4[农业科学—农药学]