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6h内对10μg ZEN分子的降解率为98%,酶活为200 U/mL,从而获得了1个新的ZEN降解酶基因.
Zearalenone(ZEN)was a mycotoxin produced mainly by fungi belonging to the genus Fusarium which frequently contaminated crops.ZEN had an intense estrogen-like effects and was extremely carcinogenic,which posed great threats to the health of animals and human.Through homologous alignment in NCBI,some homologous sequences of ZHD101 degradation enzyme were found,and one of them which came from fungus-protein sequences of Marssonina brunnea-was found to be 32%homologic to ZHD101,and the gene size was 861 bp.To determine its ZEN-degrading activity,the full-length gene was obtained by chemical synthetic method,and the protein expression was made by constructing E.coli Expression System HPLC results showed that the degradation rate of protein aganst 10 jxg ZEN within 6 h was 98%;the enzyme activity was 200 U/mL and a new ZEN-degradation enzyme was successfully obtained.
作者
柴成梁
常晓娇
王楠希
伍松陵
孙长坡
Chai Chengliang;Chang Xiaojiao;Wang Nanxi;Wu Songling;Sun Changpo(Academy of National Food and Strategic Reserves Administration,Beijing 100037)
出处
《中国粮油学报》
EI
CAS
CSCD
北大核心
2019年第S01期188-193,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).
关键词
玉米赤霉烯酮
ZEN降解酶基因
克隆
蛋白表达纯化
zearalenone
ZEN-degradation enzyme
gene cloning
protein expression and purification
作者简介
柴成梁,男,1984年出生,助理研究员,生物化学与分子生物学。;通信作者:孙长坡,男,1975年出生,研究员,粮油微生物与质量安全。
