INCREASED THERMAL-STABILITY OF PROTEINS IN THE PRESENCE OF SUGARS AND POLYOLS

Sugars and polyols stabilize proteins against heat denaturation. Scanning calorimetry was used to obtain a quantitative estimate of the degree of stabilization. Solutions of ovalbumin, lysozyme, conalbumin and .alpha.-chymotrypsinogen were heated at a constant rate, and the temperature of the maximum rate of denaturation was estimated (Tm). Addition of a sugar or polyol raised Tm. The magnitude of the stabilizing effect (.DELTA.Tm) depended on both the nature of the protein and the nature of the sugar or polyol, ranging from 18.5.degree. C for lysozyme at pH 3 in the presence of 50% (wt/wt) sorbitol to 0.degree. C for conalbumin at pH 7 in 50% glycerol solution. This stabilization is due to the effects of sugars and polyols on hydrophobic interactions. The strength of the hydrophobic interaction was measured in model system in sucrose and glycerol solutions. Sucrose and glycerol strengthened the pairwise hydrophobic interaction between hydrophobic groups, but they reduced the tendency for complete transfer of hydrophobic groups from an aqueous to a nonpolar environment. The extent of stabilization by different sugars and polyols is explained by their different influences on the structure of water. The difference between the partial molar volume of the sugar or polyol and its van der Waals volume was used as a rough quantitative measure of the structure-making or structure-breaking effect. There was a linear relationship between this quantity and .DELTA.Tm.