[P005] Poster

12:10 PM - 1:00 PM JST(3:10 AM - 4:00 AM UTC)

[P03-329] Natural Deep Eutectic Solvents for Amyloid Dissolution and Functional Recovery:A Model Study Using an Insoluble Lysozyme

○Sora Hirano¹, Tatsuya Oshima¹, Asuka Inada¹, Toshihiro Tsuruda² (1. Department of Applied Chemistry, Faculty of Engineering, University of Miyazaki (Japan), 2. Department of Hemo-Vascular Advanced Medicine, Cardi-orenal Reseach Laboratory, Faculty of Medicine, University of Miyazaki (Japan))

Keywords:

Amyloid fibrils,Deep eutectic solvents,lysozyme,protein refolding,protein solubilization

[Purpose]The formation of insoluble amyloid fibrils is a common feature of amyloidosis and other diseases. These aggregates accumulate in tissues and disturb normal function, making them difficult to remove once formed. Deep eutectic solvents (DESs), particularly natural DESs (NADESs), are emerging as biocompatible solvent systems that can modulate protein stability. In this study, a series of NADES formulations were screened for their ability to dissolve insoluble proteins, and the enzymatic activity of lysozyme after dissolution was evaluated to assess the refolding potential of these solvents.

[Method]An insoluble amyloid-like protein was prepared in vitro using hen egg-white lysozyme. The obtained aggregates were confirmed to possess amyloid structures by Congo red staining and Thioflavin T fluorescence analysis and were defined as lysozyme aggregates (LA). LA was dissolved in various deep eutectic solvents (DESs), and the solubility, secondary structure, and recovered enzymatic activity were evaluated. DMSO and urea-saturated aqueous solution were used as reference solvents.

[Results]One specific DES exhibited remarkably high solubility toward LA, approximately five times higher than that of DMSO. The recovered enzymatic activity was three to four times higher than that observed with other DESs or DMSO.

[Conclusion]These results suggest that certain DES formulations are highly effective in dissolving amyloid-like protein aggregates while promoting recovery of enzymatic activity, indicating their potential as promising solvent systems for the treatment or processing of insoluble protein aggregates.