Selective separation of beta-lactoglobulin from sweet whey using CGAs generated from the cationic surfactant CTAB.
The selective separation of whey proteins was studied using colloidal gas aphrons generated from the cationic surfactant cetyl trimethyl ammonium bromide (CTAB). From the titration curves obtained by zeta potential measurements of individual whey proteins, it was expected to selectively adsorb the major whey proteins, i.e., bovine serum albumin, alpha-lactalbumin, and beta-lactoglobulin to the aphrons and elute the remaining proteins (lactoferrin and lactoperoxidase) in the liquid phase. A number of process parameters including pH, ionic strength, and mass ratio of surfactant to protein (M(CTAB)/M(TP)) were varied in order to evaluate their effect on protein separation. Under optimum conditions (2 mmol/l CTAB, M(CTAB)/M(TP) = 0.26-0.35, pH 8, and ionic strength = 0.018 mol/l), 80-90% beta-lactoglobulin was removed from the liquid phase as a precipitate, while about 75% lactoferrin and lactoperoxidase, 80% bovine serum albumin, 95% immunoglobulin, and 65% alpha-lactalbumin were recovered in the liquid fraction. Mechanistic studies using zeta potential measurements and fluorescence spectroscopy proved that electrostatic interactions modulate only partially the selectivity of protein separation, as proteins with similar surface charges do not separate to the same extent between the two phases. The selectivity of recovery of beta-lactoglobulin probably occurs in two steps: the first being the selective interaction of the protein with opposite-charged surfactant molecules by means of electrostatic interactions, which leads to denaturation of the protein and subsequent formation and precipitation of the CTAB-beta-lactoglobulin complex. This is followed by the separation of CTAB-beta-lactoglobulin aggregates from the bulk liquid by flotation in the aphron phase. In this way, CGAs act as carriers which facilitate the removal of protein precipitate.
Fuda
Elisabeth
E
University of Reading, School of Food Biosciences, Whiteknights, P.O. Box 226, Reading, RG6 6AP, United Kingdom.
Bhatia
Divesh
D
Pyle
D L
DL
Jauregi
Paula
P
D000818:Animals; D002412:Cations; D002417:Cattle; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000737:chemistry*; D005591:Chemical Fractionation / Q000379:methods*; D005740:Gases / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007782:Lactoglobulins / Q000737:chemistry* / Q000302:isolation & purification*; D008892:Milk / Q000737:chemistry*; D013501:Surface-Active Agents / Q000737:chemistry*
D002412:Cations; D002593:Cetrimonium Compounds; D005740:Gases; D007782:Lactoglobulins; D013501:Surface-Active Agents; D000077286:Cetrimonium