玉米赤霉烯酮降解酶基因ZEN-jjm的克隆、表达及活性分析.pdfVIP

农业生物技术学报，2010 年，第 18 卷，第2 期，第225-230 页 Journal of Agricultural Biotechnology, 2010, Vol.18, No.2, 225-230 研究报告 A Letter 玉米赤霉烯酮降解酶基因(ZEN-jjm) 的克隆、表达及活性分析 1,2 2 2 2 2 2* 程波财 史文婷 罗洁 彭福中 万翠香 魏华 1 中南大学资源加工与生物工程学院, 长沙, 410083; 2 食品科学与技术国家重点实验室, 南昌大学, 南昌, 330047 * 通讯作者, hua_wei114@ 摘 要 玉米赤霉烯酮(zearalenone, ZEN)是世界上污染范围最广泛的一种镰刀霉毒素，能导致人或动物不 孕、流产等症状，损害内脏器官并促进癌细胞的生长。本文通过设计 1 对特异性引物，从粉红螺旋聚孢霉 (Gliocladiumroseum) 中克隆获得大小为795 bp 的DNA 片段(ZEN-jjm)，通过构建E.coil 原核表达载体进行 表达。经HPLC 检测证实，IPTG 诱导后的细胞裂解上清能在3 h 内完全降解液体中1 滋g/mL 的ZEN 。序列分 析结果表明 与 存在9 个碱基的差异， 编码的ZEN 降解酶与文献报道的乳糖水解 ZEN-jjm zhd101 ZEN-jjm 酶存在3 个氨基酸的差异，但ZEN 毒素降解实验证实二者活性相似。 关键词 玉米赤霉烯酮, 粉红螺旋聚孢霉, ZEN 降解酶, 基因克隆, 表达 Cloning of Zearalenone-degraded Enzyme Gene (ZEN-jjm) and its Expression and Activity Analysis Cheng Bocai 1,2 Shi Wenting 2 Luo Jie 2 Peng Fuzhong 2 Wan Cuixiang 2 Wei Hua 2* 1 School of Minerals Processing Bioengineering, Centre South University, Changsha, 410083; 2 State Key Laboratory of Food Science and Technol- ogy, Nanchang University, Nanchang, 330047 * Corresponding author, hua_wei114@ DOI: 10.3969/j.issn.1674-7968.2010.02.004 Abstract Zearalenone (ZEN) is the most widely distributed Fusariumtoxinsall over the world. ZEN can lead to a series of estrogenic effects symptoms such as infertility and abortion, damage the internal organs, and even pro- mote the cancer cells to grow. One 795 bp DNA fragment (ZEN-jjm ) from Gliocladiumroseumin charge of lacto- nohydrolase was cloned into a vector and transferred into E.coli, the degradation capabilitites of expressed protein in the supernatant of disrupted cells was verified by HPLC test. In 3 h the concentration of ZEN decreased from 1 滋g/mL to undetected limited line. Sequencing results showed that there were 9 different bases differences in the amplified gene compared with that reported gene zhd101, resulti