文献类型：期刊文献

中文题名：生物合成谷胱甘肽种间耦合ATP再生系统的构建

英文题名：BIOSYNTHESIS OF GLUTATHIONE:CONSTRUCTION OF ATP REGENERATION SYSTEM BETWEEN RECOMBINANT E.COLI AND S.CEREVISIAE

作者：李寅[1];李华钟[1];林金萍[2];陈坚[1]

机构：[1]无锡轻工大学工业生物技术教育部重点实验室,无锡214036;[2]华东理工大学生物工程学院,上海200233

年份：2001

卷号：41

期号：2

起止页码：191

中文期刊名：微生物学报

外文期刊名：Acta Microbiologica Sinica

收录：CSTPCD;;Scopus;北大核心:【北大核心2000】；CSCD:【CSCD2011_2012】；PubMed;

基金：霍英东教育基金会第六届高等院校青年教师基金项目&&

语种：中文

中文关键词：重组大肠杆菌;面包酵母;种间耦合;ATP再生系统;谷胱甘肽;生物合成

外文关键词：Recombinant Escherichia coli, Saccharomyces cerevisiae, ATP regeneration system between two microorganisms, Glutathione, Biosynthesis, Construction

摘要：利用重组大肠杆菌Ⅱ 1中的谷胱甘肽合成酶系和面包酵母WSH J7中的ATP生物合成酶系 ,构建了一个以葡萄糖为能源的种间耦合ATP再生系统。经过通透性处理的酵母细胞几乎不能消耗葡萄糖。在反应体系中添加 1mmol/LAMP和 0 0 5mmol/LNADH ,即可启动酵母的酵解途径。提高耦合系统中的葡萄糖浓度 ,可促进GSH的合成。当葡萄糖浓度为 40 0mmol/L时 ,系统内GSH浓度达到 1 0 4mmol/L(3 2 g/L)。Mg2 +缺乏时 ,耦合系统和外加ATP的非耦合系统均不能合成谷胱甘肽。耦合系统中Mg2 +与ATP形成螯合物 ,可能是导致耦合系统中GSH产量较低的原因。在耦合系统中补加Mg2 +,反应 6h时GSH浓度达到 1 4 3mmol/L(4 4g/L)。
An ATP regeneration system between two microorganisms,in which recombinant E.coli Ⅱ 1 and \%S.cerevisiae\% WSH\|J702 were used as the biosynthesis enzymes source of glutathione and ATP,respectively,was constructed by using glucose as an energy substrate.Yeast cells after permeable treatment could hardly consume glucose.The glycolysis pathway of yeast was started by adding 1.0mmol/L AMP and 0 05mmol/L NADH simultaneously.GSH biosynthesis in the coupling system was improved with increasing glucose concentration to 400mmol/L,where the concentration of GSH reached 10 4mmol/L (3 2g/L).Both the coupling system and the non\|coupling system with ATP added could hardly produce GSH under the absence of magnesium.The low efficiency of ATP regeneration system constructed was probably caused by the chelation between magnesium and ATP.By adding magnesium to the coupling system,a final concentration of GSH,14 3mmol/L (4 4g/L),was achieved at 6h of reaction.