Paul S, Chaudhuri T K
Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, Hauz Khas, New Delhi, India.
J Appl Microbiol. 2008 Jan;104(1):35-41. doi: 10.1111/j.1365-2672.2007.03519.x.
To investigate the factors affecting expression and solubilization of Escherichia coli maltodextrin glucosidase in E. coli.
Expression level and solubilization of the recombinant E. coli maltodextrin glucosidase was studied in E. coli at different temperatures, in presence of overexpressed GroEL, GroES and externally supplemented glycerol. Aggregation of maltodextrin glucosidase in the cytoplasm was partially prevented by the co-expression of GroEL and GroES, and using externally supplemented glycerol or lowering the culture temperature. Co-expression of GroEL and GroES or simultaneous presence of overexpressed GroEL, GroES and externally supplemented glycerol together resulted significant increase of the activity of maltodextrin glucosidase. The growth rate of E. coli was inhibited by the formation of inclusion bodies whereas the presence of overexpressed GroEL, GroES alone or together with glycerol enhanced the growth rate of E. coli substantially.
The results indicated that lowering the temperature, use of GroEL, GroES and glycerol could be few controlling factors for the solubilization of recombinant aggregation-prone maltodextrin glucosidase in E. coli.
Our study could help in developing the strategy for enhancing the production of soluble industrial enzymes and finding the therapeutic agents against protein misfolding diseases.
研究影响大肠杆菌中麦芽糖糊精葡萄糖苷酶表达和溶解的因素。
在不同温度下，在过表达GroEL、GroES以及外部添加甘油的情况下，研究重组大肠杆菌麦芽糖糊精葡萄糖苷酶在大肠杆菌中的表达水平和溶解性。通过共表达GroEL和GroES、使用外部添加的甘油或降低培养温度，可部分防止麦芽糖糊精葡萄糖苷酶在细胞质中聚集。GroEL和GroES的共表达或过表达的GroEL、GroES与外部添加甘油同时存在，均导致麦芽糖糊精葡萄糖苷酶活性显著增加。包涵体的形成抑制了大肠杆菌的生长速度，而过表达的GroEL、GroES单独或与甘油一起存在则显著提高了大肠杆菌的生长速度。
结果表明，降低温度、使用GroEL、GroES和甘油可能是控制重组易聚集麦芽糖糊精葡萄糖苷酶在大肠杆菌中溶解的几个因素。
我们的研究有助于制定提高可溶性工业酶产量的策略，并寻找针对蛋白质错误折叠疾病的治疗药物。
