Effects of hydroxypropyl cyclodextrins on the reactivation of SDS-denatured aminoacylase.

Cyclodextrins are natural-occurring circular oligosaccharides with an internal hydrophobic cavity and external hydrophilic edges. Because cyclodextrins bind with protein aromatic residues, they can prevent protein aggregation, and their ability to bind with detergents enables them to act as stripping reagents to release proteins from protein-detergent complexes. In this research, we investigated the effects of three hydroxypropyl cyclodextrins (HPCDs) on the refolding of aminoacylase from SDS-denatured states. It was found that the three HPCDs could effectively assist aminoacylase reactivation though they have different abilities. HP-gamma-CD, which has the largest cavity among the three HPCDs, was the most efficient one. Spectroscopic results further indicated that the secondary structure recovery of aminoacylase could be completed with the help of low concentrations of HPCDs. However, the activity of the released protein could not fully recover even though high concentrations of HPCDs were used. The concentration-dependent effects of HPCDs also indicated that cyclodextrins could also act as folding assistants in addition to acting as stripping reagents during the refolding of detergent-denatured proteins.