Effect of polyols and sugars on heat-induced flavin dissociation in glucose oxidase

A thermodynamic investigation was carried out on heat-induced flavin dissociation in Aspergillus niger glucose oxidase. Experimental measurements performed by difference spectroscopy showed that the dissociation of the FAD cofactors is a highly cooperative process and is probably related to the extended conformational changes resulting from protein unfolding. Microenvironmental modifications attained by the addition of polyhydric compounds (glycerol, fructose, sucrose and sorbitol) from 10 to 30% by weight were found to hinder the dissociation. The stabilizing effect provided by these substances was interpreted as a consequence of preferential exclusion phenomena, which are likely to be determined by the perturbation of the surface tension of water, in the case of sugars, or by the solvophobic effect, in the case of glycerol.