Bioprotection Can Be Tuned with a Proper Protein/Saccharide Ratio: The Case of Solid Amorphous Matrices.

Saccharides, and in particular trehalose, are well known for their high efficiency in protecting biostructures against adverse environmental conditions. The protein dynamics is known to be highly inhibited in a low-water trehalose host medium, the inhibition being markedly dependent on the amount of residual water. Besides hydration, the protein/sugar ratio is expected to affect the properties of saccharide amorphous matrices. In this work, we report an infrared spectroscopy study in dry amorphous matrices of various sugars (the disaccharides trehalose, maltose, sucrose, and lactose, and the trisaccharide raffinose) containing myoglobin, at different protein/sugar ratios. We analyze the stretching band of the bound CO molecule and the water association band. Such bands have already been successfully exploited for the simultaneous study of thermal evolution of a matrix and embedded protein. The results show a high dependence of protein and matrix signals on the protein/sugar ratio, the system behavior evolving from situations where (i) the protein slaves the matrix to (ii) protein ↔ matrix coupling/uncoupling, then to (iii) the matrix slaving the protein, with increasing sugar concentration. This supports a mutual protein ↔ matrix structural and dynamic influence in low hydrated systems, indicating that the protein/solvent master and slave paradigm does not strictly hold, but the mutual relationship depends on the relative concentrations. Furthermore, for each sugar, an optimal protein/sugar concentration ratio can be identified, which maximizes the protein preservation; under such a condition, the water content is minimal.