Method for enhancing solubility of the expressed recombinant proteins in Escherichia coli.
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.
Ghosh
Sudip
S
National Institute of Nutrition (ICMR), Hyderabad, India.
Rasheedi
Sheeba
S
Rahim
Sheikh Showkat
SS
Banerjee
Sharmistha
S
Choudhary
Rakesh Kumar
RK
Chakhaiyar
Prachee
P
Ehtesham
Nasreen Z
NZ
Mukhopadhyay
Sangita
S
Hasnain
Seyed E
SE
D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D006033:Glycylglycine; D002479:Inclusion Bodies; D009169:Mycobacterium tuberculosis / Q000235:genetics*; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry; D017422:Sequence Analysis, DNA; D012995:Solubility
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D006033:Glycylglycine