Ghosh Sudip, Rasheedi Sheeba, Rahim Sheikh Showkat, Banerjee Sharmistha, Choudhary Rakesh Kumar, Chakhaiyar Prachee, Ehtesham Nasreen Z, Mukhopadhyay Sangita, Hasnain Seyed E
National Institute of Nutrition (ICMR), Hyderabad, India.
Biotechniques. 2004 Sep;37(3):418, 420, 422-3. doi: 10.2144/04373ST07.
The production of correctly folded protein in Escherichia coli is often challenging because of aggregation of the overexpressed protein into inclusion bodies. Although a number of general and protein-specific techniques are available, their effectiveness varies widely. We report a novel method for enhancing the solubility of overexpressed proteins. Presence of a dipeptide, glycylglycine, in the range of 100 mM to 1 M in the medium was found to significantly enhance the solubility (up to 170-fold) of the expressed proteins. The method has been validated using mycobacterial proteins, resulting in improved solubilization, which were otherwise difficult to express as soluble proteins in E. coli. This method can also be used to enhance the solubility of other heterologous recombinant proteins expressed in a bacterial system.
由于在大肠杆菌中过表达的蛋白质会聚集形成包涵体，因此产生正确折叠的蛋白质通常具有挑战性。尽管有许多通用的和针对特定蛋白质的技术，但它们的有效性差异很大。我们报告了一种提高过表达蛋白质溶解度的新方法。发现在培养基中存在浓度范围为100 mM至1 M的二肽甘氨酰甘氨酸可显著提高表达蛋白质的溶解度（高达170倍）。该方法已通过分枝杆菌蛋白进行了验证，从而提高了溶解性，否则这些蛋白很难在大肠杆菌中作为可溶性蛋白表达。该方法还可用于提高在细菌系统中表达的其他异源重组蛋白的溶解度。
