Micro-Raman spectroscopic observation of water expulsion induced destruction of hydrophobic clusters in crystalline lysozyme

In this paper, three kinds of solid lysozyme samples with different water contents were investigated by confocal Raman spectroscopy. For the rod-like lysozyme crystal with highest water content, a sudden decrease of the intensity ratio of the doublet at 1338 and 1360 cm^-1 was observed when the ambient relative humidity (RH) was lower than 86%, indicating the destruction of hydrophobic clusters of lysozyme induced by the expulsion of the hydration water from the crystal. In contrast to the rod-like crystal, tetragonal crystal and floor-like precipitate with a smaller amount of water showed no change of the structures of the hydrophobic clusters when the relative humidity was decreasing. The presence of bulk water in the rod-like crystal is believed a necessary factor for the function of the hydration water which promotes the hydrophobicity of hydrophobic clusters.