Refolding of an unstable lysozyme by gradient removal of a solubilizer and gradient addition of a stabilizer
Authors: Kohyama, Keisuke; Matsumoto, Toshihiko; Imoto, Taiji
Source: The Journal of Biochemistry, Volume 147, Number 3, 6 March 2010 , pp. 427-431(5)
Publisher: Oxford University Press
Abstract:Earlier, we formally established an effective refolding procedure for a protein by gradient removal of a solubilizer such as urea [Maeda et al. (1995) Effective renaturation of reduced lysozyme by gentle removal of urea. Protein Eng. 8, 201205]. However, this procedure was less effective for unstable proteins. We developed here an excellent method to add protein stabilizer so as to get reasonable amounts of folded protein under the concentration of solubilizer where the unstable protein does not form aggregate. We examined many stabilizers and found that 60 of a concentrated (2.5 mg/ml) unstable protein can be refolded using 40 glycerol as the best stabilizer. This procedure can be widely applicable for the refolding of unstable proteins.
Document Type: Regular Paper
Publication date: 2010-03-06