Separation of beta-lactoglobulin A, B and C variants of bovine whey using capillary zone electrophoresis.
beta-Lactoglobulin is a whey protein that affects milk composition and product functionality and which can be present in up to eight genetic variant forms. A free zone capillary electrophoresis method has been developed to separate and identify the beta-lactoglobulin A, B and C variants. Three buffer systems [borate, 2-(N-morpholino)-ethanesulphonic acid (MES) and bis(2-hydroxyethyl)imino-tris(hydroxymethyl)methane (Bistris)] were examined over a range of pH values and with the addition of the separation buffer modifiers Tween 20 and/or ethanolamine. The most successful combination of these was 50 mM MES at pH 8.0 with the addition of 0.1% Tween 20 which clearly resolved the three variants from both each other and from the other whey proteins even though the MES buffer was acting well outside its pKa range (pH 5.3-7.3). The retention times and identification of the individual variants were verified by spiking with commercially purified beta-lactoglobulin A and B proteins and a beta-lactoglobulin AC whey. The method was then used to phenotype beta-lactoglobulin in a sample population of New Zealand Jersey cows.
Paterson
G R
GR
Food Science Section, New Zealand Dairy Research Institute, Palmerston North.
Hill
J P
JP
Otter
D E
DE
D000595:Amino Acid Sequence; D000818:Animals; D002197:Capillary Action; D002417:Cattle; D004586:Electrophoresis / Q000379:methods*; D005260:Female; D006863:Hydrogen-Ion Concentration; D007782:Lactoglobulins / Q000737:chemistry / Q000302:isolation & purification*; D008894:Milk Proteins / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D000067816:Whey Proteins
D007782:Lactoglobulins; D008894:Milk Proteins; D000067816:Whey Proteins