【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)。更多还原

【Abstract】 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.更多还原