酿酒酵母中表达神经肉毒素A及其功能研究  

Expression of Botulinum Neurotoxins A in Saccharomyces cerevisiae and Its Functional Research

作　　者：NDIKUBWIMANA Jean de Dieu 邵侃凯 高晓冬[1] 中西秀树[1] SHAO Kankai;GAO Xiaodong;NAKANISHI Hideki(School of Biotechnology,Jiangnan University,Wuxi 214122,China)

机构地区：[1]江南大学生物工程学院,江苏无锡214122

出　　处：《食品与生物技术学报》2019年第2期87-92,共6页Journal of Food Science and Biotechnology

基　　金：国家"111计划"项目(111-2-06);教育部科学研究项目(313027);高等学校博士学科点专项科研基金项目(20120093110001);江南大学自主科研项目(JUSRP311A02)

摘　　要：肉毒神经毒素(BoNT)是肉毒梭菌产生的神经毒素,是最强大的毒素之一。为了研究肉毒神经毒素A在酵母中的活性及功能,在酿酒酵母细胞中表达BoNT/A LC,观察其对酵母生长影响以及通过蛋白免疫印迹方法研究其在酵母中蛋白表达水平。结果表明:BoNT/A抑制野生型酵母的生长;在酿酒酵母细胞中BoNT/A具有水解酶活性,当其活性位点突变后,BoNT/A突变体的表达不会抑制酿酒酵母的生长;过量表达酵母的SNAP25同源基因SEC9可以回补BoNT/A轻链过表达的酵母的生长。在BoNT/A轻链表达的酵母细胞中,其催化活性可以通过酵母的生长来检测。因此,这类特殊的酵母细胞可以用来分析BoNT/A。Botulinum toxin(BoNT) is a neurotoxin produced by Clostridium botulinum,which is one of the most powerful toxin. To study the activity and function of BoNT/A in yeast,BoNT/A LC in yeast is expressed,observing the yeast growth and BoNT/A protein expressing level by western blot. Expression of LC of BoNT cerotype A caused a growth defect in yeast cells,and the toxicity is due to the hydrolytic activity. It is because catalytically inactive BoNT/A LC does not cause a growth defect in yeast cells. Overexpression of a yeast SNAP-25 homologue gene,SEC9,could rescue the growth defect in BoNT/A LC expressing cells. The results demonstrate that,in BoNT/A LC expressing yeast cells,the hydrolytic activity can be monitored through their growth. Thus,such cells can be used to analyze BoNT/A.

关 键 词：肉毒神经毒素A(BoNT/A) 酿酒酵母 SEC9 

分 类 号：Q786[生物学—分子生物学]