S. Bouhallab et al., Formation of stable covalent dimer explains the high solubility at pH 4.6 of lactose-beta-lactoglobulin conjugates heated near neutral pH, J AGR FOOD, 47(4), 1999, pp. 1489-1494
The solubility of lactose-beta-lactoglobulin conjugates at pH 4.6, after he
ating near neutral pH in phosphate buffer/0.116 M NaCl, was investigated by
size exclusion chromatography and compared with unmodified protein. Heated
conjugates in the temperature range 65-90 degrees C showed greater solubil
ity at pH 4.6. The proportion of soluble protein increased with the number
of bound lactose molecules. Total solubility was obtained for conjugates wi
th nine lactose residues attached per monomer of beta-lactoglobulin. The pr
otective effect of bound sugar toward precipitation was associated with the
formation of soluble disulfide cross-linked dimers, highly accessible to t
rypsin digestion. These results suggested that bound lactose, through steri
c hindrance and high surface hydrophilicity, prevents the thiol-disulfide e
xchange reactions of the polymerization-aggregation process of lactose-beta
-lactoglobulin conjugates.