Purification process for the preparation and characterizations of hen egg white ovalbumin, lysozyme, ovotransferrin, and ovomucoid.
In this study, four major egg white proteins were purified by two step ion exchange chromatography followed by precipitation. Lysozyme and ovalbumin were separated with Q Sepharose Fast Flow anion exchange chromatography in the first step, resulting in two peaks of lysozyme and three peaks of ovalbumin with 87% and 70% purity by HPLC, respectively. Ovotransferrin was separated with CM-Toyopearl 650 M cation exchange chromatography in the second step, giving 80% purity. Ovomucoid was precipitated from the partial purified protein fraction from the first two steps, and concentrated in the final step to yield 90% purity. Protein recoveries were estimated to be 55, 21, 54, and 21% for lysozyme, ovotransferrin, ovalbumin, and ovomucoid, respectively. Lysozyme was identified from the purified peaks using zymogram refolding gel, whereas ovalbumin was identified by western blotting. Purified ovotransferrin and ovomucoid was identified by mass spectrometry. The results indicate that this purification process is adequate for preparation of lysozyme, ovalbumin, ovotransferrin, and ovomucoid, at least on a laboratory scale.
Tankrathok
Anupong
A
Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen, Thailand.
Daduang
Sakda
S
Patramanon
Rina
R
Araki
Tomohiro
T
Thammasirirak
Sompong
S
D000818:Animals; D005591:Chemical Fractionation / Q000379:methods*; D002645:Chickens; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004527:Egg Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D004529:Egg White / Q000737:chemistry*
D004527:Egg Proteins