Correlation between thermal aggregation and stability of lysozyme with salts described by molar surface tension increment: an exceptional propensity of ammonium salts as aggregation suppressor.

Protein aggregation is a critical problem for biotechnology and pharmaceutical industries. Despite the fact that soluble proteins have been used for many applications, our understanding of the effect of the solution chemistry on protein aggregation still remains to be elucidated. This paper investigates the process of thermal aggregation of lysozyme in the presence of various types of salts. The simple law was found; the aggregation rate of lysozyme increased with increasing melting temperature of the protein (T (m)) governed by chemical characteristics of additional salts. Ammonium salts were, however, ruled out; the aggregation rates of lysozyme in the presence of the ammonium salts were smaller than the ones estimated from T (m). Comparing with sodium salts, ammonium salts increased the solubility of the hydrophobic amino acids, indicating that ammonium salts adsorb the hydrophobic region of proteins, which leads to the decrease in aggregation more effectively than sodium salts. The positive relation between aggregation rate and T (m) was described by another factor such as the surface tension of salt solutions. Fourier transform infrared spectral analysis showed that the thermal aggregates were likely to form beta-sheet in solutions that give high molar surface tension increment. These results suggest that protein aggregation is attributed to the surface free energy of the solution.