Isolation and characterization of copolymers of beta-lactoglobulin, alpha-lactalbumin, kappa-casein, and alphas1-casein generated by pressurization and thermal treatment of raw milk.
Raw skim milk was submitted to high pressure (300 to 600 MPa) and temperature (4 to 70 degrees C) treatments for 2 or 5 min. The combined effects of pressure and temperature on milk proteins induced structural changes and polymer and copolymer formation characterized by anion-exchange and size-exclusion fast protein liquid chromatography and electrophoretic techniques. Approximately half of the beta-lactoglobulin formed polymers, and the other half formed large copolymers, mainly with kappa-casein, alpha-lactalbumin via intermolecular disulfide bond exchange, and alpha(s1)-casein via physicochemical interactions, in proportions of 1.0:0.7:0.3:0.1, respectively. Minor whey proteins (serum albumin, immunoglobulins, and lactoferrin) also participated in the formation of the copolymers but to a lesser extent. Two populations of the copolymers were found with apparent molecular masses ranging from 440 to 2000 kDa for the first and more than 2000 kDa for the second. On the contrary, for heated milks the aggregation kinetics obtained by combination of high pressure and thermal treatment were very fast, as no intermediates such as dimers and small size oligomers were observed after pressurization, whatever the temperature studied. Lactosylation of proteins as well as proteolysis were very limited. A beta-casein amino-terminal peptide of 22 kDa was specifically recovered in milk samples treated under the more drastic conditions (500 MPa/55 degrees C per 5 min and 600 MPa/70 degrees C per 5 min) and might have been generated by neutral proteases such as elastase released from somatic cells present in milk. No casein was released from the micelle whatever the combination of high pressure and temperature studied.
Laboratoire de Sciences Animales, U.S.C. INRA no. 12340, ENSAIA, Institut National Polytechnique de Lorraine, Vandoeuvre-lès-Nancy, France.
M A
MA
Nabhan
J-M
JM
Girardet
S
S
Campagna
J-L
JL
Gaillard
Y
Y
Le Roux
D000595:Amino Acid Sequence; D000818:Animals; D002364:Caseins / Q000032:analysis / Q000302:isolation & purification*; D005591:Chemical Fractionation; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006358:Hot Temperature; D007768:Lactalbumin / Q000032:analysis / Q000302:isolation & purification*; D007782:Lactoglobulins / Q000032:analysis / Q000302:isolation & purification*; D008892:Milk / Q000737:chemistry*; D011108:Polymers; D011312:Pressure
D002364:Caseins; D007782:Lactoglobulins; D011108:Polymers; D007768:Lactalbumin