A simplified purification procedure of alpha-lactalbumin from milk using Ca(2+)-dependent adsorption in hydrophobic expanded bed chromatography.
The technique of expanded bed adsorption is originally designed for a direct recovery of proteins from fermentor feedstocks. In this article we describe the use of expanded bed adsorption for the recovery of alpha-lactalbumins from defatted milk using the hydrophobic Streamline Phenyl gel. alpha-Lactalbumins are Ca(2+)-binding proteins. Upon Ca2+ removal, they undergo a significant conformation change rendering them more hydrophobic. Based on this unique property we develop a protocol for fast and efficient purification of alpha-lactalbumin from milk. The use of this technique results in a reduction of the number of chromatographic purification steps.
Noppe
W
W
Interdisciplinary Research Centre, Katholieke Universiteit Leuven, Kortrijk, Belgium. wim.noppe@kulak.ac.be
Haezebrouck
P
P
Hanssens
I
I
De Cuyper
M
M
D000327:Adsorption; D000818:Animals; D002118:Calcium / Q000737:chemistry*; D002850:Chromatography, Gel / Q000379:methods*; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D007768:Lactalbumin / Q000302:isolation & purification*; D008892:Milk / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet
D007768:Lactalbumin; D002118:Calcium