Ep. Schokker et al., Characterization of intermediates formed during heat-induced aggregation of beta-lactoglobulin AB at neutral pH, INT DAIRY J, 9(11), 1999, pp. 791-800
Heat-induced aggregation of beta-lactoglobulin AB (17 mg mL(-1) in 0.005 M
NaCl; pH 7.0; 78.5 degrees C) was studied using size-exclusion chromatograp
hy in combination with multi-angle laser light scattering and electrophoret
ic techniques. By choosing the chromatographic condition such that the nati
ve beta-lactoglobulin was partially associated, the formation of irreversib
ly altered monomers and non-native dimers could be monitored. Heat-induced
aggregation of beta-lactoglobulin occurred via many intermediates, and main
ly via disulphide bonding and to a much lesser extent via non-covalent inte
ractions. In the early stages of the aggregation of beta-lactoglobulin non-
native dimers and oligomers were formed, presumably mainly via thiol-disulp
hide exchange and to a lesser extent via thiol-thiol oxidation and non-cova
lent interactions. In later stages aggregates with a continuous distributio
n of molar masses from 10(5) to 2 x 10(6) Da were formed, presumably via th
e incorporation of monomers and small aggregates into larger aggregates mai
nly through disulphide bonding. (C) 2000 Elsevier Science Ltd. All rights r
eserved.