热休克蛋白增加大肠杆菌抗逆性和乙醇产量的研究  

Study of Using Hsp in E.coli to Increase Its Resistance to Adversituy and Its Production of Ethanol

作　　者：李伟丽[1] 李海燕[1] 李良[1] 曾一梅[1] 汪浩勇[1] 

机构地区：[1]湖北工业大学,湖北武汉430068

出　　处：《生物技术》2008年第5期54-58,共5页Biotechnology

基　　金：湖北省优秀中青年团队计划项目("构建乙醇高温发酵基因工程菌";T200705)资助

摘　　要：目的:研究热休克蛋白对增加大肠杆菌抗逆性和乙醇产量的影响。方法:运用基因工程技术,用大肠杆菌的Lac启动子、运动发酵单胞菌的丙酮酸脱羧酶基因(pdc)和乙醇脱氢酶基因(adhB),构建可以在大肠杆菌中表达的Lac-AP操纵子。Lac-AP操纵子导入大肠杆菌,可使大肠杆菌发酵糖生产乙醇。再用来自超嗜热菌强烈火球菌(Pyrococcus furiosus)的小分子热休克蛋基因(sHsp),构建在大肠杆菌中表达的Lac-APH操纵子。结果:成功地构建了耐高温产生乙醇的大肠杆菌LAPH和LAP,它们发酵后乙醇的产量分别为11.5g/L、7.9g/L,而对照菌LH的产量只有为0.5g/L。与对照菌LH相比,LAPH和LAP的产量分别提高了23倍和15.8倍。结果证明:与对照LAP相比,热休克蛋白使菌种LAPH的45℃温度耐受性提高15.75倍、50℃温度耐受性提高40.7倍,乙醇的产量增高高达4.74倍。结论:研究表明,小分子热休克蛋白的表达,可使细菌在致死温度下的存活率显著提高,耐受温度明显增强,乙醇产量显著提高。Objective： sHsp could be used to increase the resistance to adversity and production of ethanol in Escherichia coli. Method： The Lac - AP operon which could be expressed in E. coli to make E. coli to produce alcohol was constructed by using pdc and adhB from Zymomonas mobilis under the control of Lac promoter. The small heat shock protein gene （sHsp） from Pyrococcus furiosus was added to the Lac - AP to give rise to the Lac- APH operon. Resttlt：Genetic engineered Escherichia coli IMP and IAPH for producing fuel alcohol bad been constructed, the produetion of ethanol fermented by them ware respectively up to 11.5g/L,7.9g/L, but the production of the contrast strain LH was only 0.5g/L. The results show that, compared with contrast strain LAP, the temperature tolerance of strain LAPH had increased 15.75 times in 45℃ and 40.7 times in 50℃, and the production of ethanol had increased 4.74 times by the expression of sHsp. Conclusion：This study shows that the expression of sHsp can dramatically increase the tolerance of bacterium to the high temperature, can dramatically increase the alcohol production of bacterium even under its lethal stress conditions.

关 键 词：丙酮酸脱羧酶基因(pdc) 乙醇脱氢酶基因(adhB) 大肠杆菌 燃料乙醇 热休克蛋白基因(sHsp) 

分 类 号：Q785[生物学—分子生物学]