Modulation of thermal stability and heat-induced gelation of β-lactoglobulin by high glycerol and sorbitol levels

The influence of glycerol and sorbitol on the thermal stability and heat-induced gelation of beta-lactoglobulin (beta-lg) in aqueous solutions was investigated. The thermal stability of beta-lg was characterized by measuring the thermal denaturation temperature (T-m) using differential scanning calorimetry, while its gelation properties were characterized by measuring the gelation temperature (T-gel) and final gel rigidity (G*) using dynamic shear rheology. All experiments were carried out using aqueous solutions containing 10% (w/w) beta-lg, glycerol (0-70% w/w) or sorbitol (0-55% w/w), and 5 mM phosphate buffer (pH 7.0). No salt was added to these solutions so that there was a relatively strong electrostatic repulsion between the protein molecules, which usually prevents gelation. When the cosolvent concentration was increased from 0% to 50%, T-m increased from 74 to 86 degrees C for sorbitol, but only from 74 to 76 degrees C for glycerol, which indicated that sorbitol was much more effective at stabilizing the native state of the globular protein than glycerol. Protein solutions containing sorbitol (0-55%) did not form a gel after heating, but those containing glycerol formed gels when the cosolvent concentration exceeded about 10%, with G* increasing with increasing glycerol concentration. We attribute these effects to differences in the preferential interactions of polyols and water with the surfaces of native and heat-denatured proteins, and their influence on the protein-protein collision frequency. (c) 2006 Elsevier Ltd. All rights reserved.