文献类型：期刊文献

中文题名：分子伴侣GroEL对青霉素G酰化酶亚基折叠的影响

英文题名：Effect of GroEL Chaperone on Posttranslational Process of Penicillin G Acylase Gene in E.coli

作者：徐京宁[1];杨运桂[2];龚毅[2];杨胜利[2];俞俊棠[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海200237;[2]中国科学院上海生物工程研究中心,上海200233

年份：2002

卷号：28

期号：2

起止页码：168

中文期刊名：华东理工大学学报（自然科学版）

外文期刊名：Journal of East China University of Science and Technology

收录：CSTPCD;;EI(收录号：2002397116939);Scopus;北大核心:【北大核心2000】；CSCD:【CSCD2011_2012】；

语种：中文

中文关键词：青霉素G酰化酶;翻译后加工;分子伴侣;GroEL;宿主

外文关键词：penicillin G acylase; posttranslational process; molecular chaperon (GroEL); host

摘要：采用 Tac启动子控制表达质粒 ,在不同的宿主细胞中表达了青霉素 G酰化酶 ( PAC)。检测这些菌株所表达的 PAC活性 ,分析细胞内分子伴侣 Gro EL含量、PAC翻译后加工为 α、β亚基的状况 ,以及它们之间的关系。结果表明 :质粒 p KK- SP在不同宿主中表达时 ,翻译后加工状况有明显差异 ,单位质量细胞所表达的 PAC活性与翻译后加工效率相关 ,且与细胞内分子伴侣 Gro EL在菌体总蛋白中含量正相关。同时也阐明了亚基的折叠成为翻译后加工过程的限制步骤 ,细胞内分子伴侣 Gro EL有助于
The expression level of penicillin G acylase (PAC) gene in different host strains harbouring same plasmid (pKK SP) were different, among them, the highest expression was achieved in E.coli BL21(DE3), the lowest in E.coli BL21 and moderate expression levels were observed in E.coli D816 and E.coli HB101. The special activity of penicillin G acylase gene in four host strains are all dependent on molecular chaperon GroEL content in these cells. Analysis of results from sodim dodecyl sulfate/polyacrylamide gel electrophoresis and immuno blot of whole cell proteins in these cells indicated that folding of subunits were bottleneck steps limiting posttranslational process of penicillin G acylase. The posttranslational efficiency (folding rate, secretion rate) of PAC is dependent on molecular chaperon GroEL content in these cells.