earticle

영어

The expression of recombinant proteins in microorganisms frequently leads to the formation of insoluble aggregates, inclusion bodies (IBs). Thus, an additional protein unfolding and refolding process is required to convert the IBs to the water-soluble active proteins. Various techniques, such as batch-dilution, dialysis and chromatography-based refolding processes, have been reported for this. In addition, numerous additives such as amino acids (cysteine, arginine, lysine and proline), plolyamine (putrescine, spermidine and spermine) and oligosaccharides have been suggested to enhance the refolding efficiency. Especially, in the case of refolding of disulfide bond-containing proteins, the redox reagents (cysteine, glutathione and cysteamine) are added to recover disulfide bonds. Room temperature ionic liquids (RTILs) are organic salts which do not crystallize at room temperature. Tunable hydrophobicity and polarity of RTILs leads the expansion of its applications in chemical and biological processes. Recently, RTILs were recommended as a promising additive of protein refolding. In this study, the effect of hydrophobicity and sulfur residue in RTILs combined with redox system on the dilution-based refolding of protein was investigated. The RTILs which has MS cation was more effective than BF4. The refolding yield was proportionally decreased with increased hydrophobicity, which has longer alkyl chain length, of RTILs. And without redox system, there are no significant
additive effect on lysozyme refolding efficiency.