玉米赤霉烯酮降解酶基因mbZHD的原核表达及其降解毒素初步研究  

Preliminary Study on Prokaryotic Expression and Its Degradation Activity of a New Zearalenone-Degradation Enzyme(mbZHD)

作　　者：柴成梁[1] 常晓娇[1] 王楠希 伍松陵[1] 孙长坡[1] 

机构地区：[1]国家粮食局科学研究院,北京100037

出　　处：《中国粮油学报》2017年第8期29-33,38,共6页Journal of the Chinese Cereals and Oils Association

基　　金：国家科技支撑计划(2015BAK43B01);公益性行业(粮食)科研专项(201513006)

摘　　要：玉米赤霉烯酮(Zearalenone,ZEN)是由镰刀菌属产生的次生代谢物,是极易污染粮食作物的真菌毒素之一。ZEN具有强烈的雌激素样效应及致癌性,给人类和动物的健康造成巨大威胁。本研究在NCBI数据库中进行同源比对,获得了一些ZHD101降解酶的同源基因,其中,1个来源于真菌——杨盘二孢菌(Marssonina brunnea)的蛋白序列与ZHD101具有32%的同源性,基因大小为861 bp。通过化学合成方法得到该全长基因,通过构建E.coli原核表达系统进行蛋白表达,蛋白经亲和色谱纯化后对ZEN分子进行降解活性验证,HPLC结果表明,该蛋白在6 h内对10μg ZEN分子的降解率为98%,酶活为200 U/m L,从而获得了1个新的ZEN降解酶基因。Zearalenone （ZEN） is a mycotoxin produced mainly by fungi belonging to the genus Fusarium which frequently contaminates crops. ZEN is a serious threat to vertebrate animals including humans because it is frequently implicated in reproductive disorders of farm animals and in hyperoestrogenic syndromes and carcinogenicity in humans. Through BLAST in NCBI Using ZHD101 as template, some homologous sequences were found, and one of the possible ZEN-degrading genes named mbZHD from Marssonina brunnea was decided to further study. To determine its ZEN-degrading activity, the gene fragment was synthesized chemically and cloned into the E.coli Expression System, the protein was then purified using Ni-NTA affinity chromatography. The protein was incubated with ZEN for 6 h, and then analyzed by high-performance liquid chromatography system（HPLC）. The result indicated that the enzyme activity is 200 U/mL and a new ZEN-degradation enzyme was successfully obtained.

关 键 词：玉米赤霉烯酮ZEN降解酶基因 克隆蛋白表达纯化 

分 类 号：Q71[生物学—分子生物学]