热休克蛋白对枯草芽孢杆菌抗逆性和乙醇产量的影响

The Study of Using Hsp in Genetic Engineered Bacillus subtilis to Increase its Resistance to Adversity and its Production of Ethanol

[目的]研究热休克蛋白对增强枯草芽孢杆菌的抗逆性和增加乙醇产量的影响。[方法]运用基因工程技术,用大肠杆菌的Lac启动子、运动发酵单胞菌丙酮酸脱羧酶基因(pdc)和乙醇脱氢酶基因(adhB),构建在枯草芽孢杆菌中表达的BLAP操纵子,BLAP操纵子导入枯草芽孢杆菌可使其发酵糖生产乙醇。然后用超嗜热菌强烈火球菌(Pyrococcusfuriosus)的小分子热休克蛋白基因(sHsp)构建在枯草芽孢杆菌中表达的BLAPH操纵子。[结果]成功地构建了能表达小分子热休克蛋白,产生乙醇的枯草芽孢杆菌。[结论]小分子热休克蛋白的表达,使枯草芽孢杆菌在致死温度下的存活率显著提高,对温度及乙醇的耐受性显著增强。同时枯草芽孢杆菌的乙醇产量显著提高。

[Objective] sHsp could be used to increase the resistance to adversity and production of ethanol in Bacillus subtilis.[Methods]In this study, BLAP operon which could be expressed in Bacillus subtilis to make it 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 BLAP to give rise to the BLAPH operon.[Results]Genetic engineered Bacillus subtilis for producing fuel alcohol had been constructed.[Conclusion]This study shows that the expression of the small heat shock protein gene can dramatically increase the tolerance of Bacillus subtilis to the high temperature and high alcohol concentration, can dramatically increase the alcohol production in bacterium even under its lethal stress conditions.