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http://dx.doi.org/10.5851/kosfa.2016.36.6.769

An Efficient Method for Co-purification of Eggshell Matrix Proteins OC-17, OC-116, and OCX-36  

Zhang, Maojie (National and Local Joint Engineering Research Center for Egg Processing Technology, College of Food Science and Technology, Huazhong Agricultural University)
Wang, Ning (National and Local Joint Engineering Research Center for Egg Processing Technology, College of Food Science and Technology, Huazhong Agricultural University)
Xu, Qi (National and Local Joint Engineering Research Center for Egg Processing Technology, College of Food Science and Technology, Huazhong Agricultural University)
Harlina, Putri Widyanti (National and Local Joint Engineering Research Center for Egg Processing Technology, College of Food Science and Technology, Huazhong Agricultural University)
Ma, Meihu (National and Local Joint Engineering Research Center for Egg Processing Technology, College of Food Science and Technology, Huazhong Agricultural University)
Publication Information
Food Science of Animal Resources / v.36, no.6, 2016 , pp. 769-778 More about this Journal
Abstract
In this study, we improved the eggshell-membrane separation process by separating the shell and membrane with EDTA solution, evaluating effects of three different extraction solutions (acetic acid, EDTA, and phosphate solution), and co-purifying multiple eggshell proteins with two successive ion-exchange chromatography procedures (CM Sepharose Fast Flow and DEAE Sepharose Fast Flow). The recovery and residual rates of eggshell and membrane separated by the modified method with added EDTA solution were 93.88%, 91.15% and 1.01%, 2.87%, respectively. Ovocleidin-116 (OC-116) and ovocalyxin-36 (OCX-36) were obtained by loading 50 mM Na-Hepes, pH 7.5, 2 mM DTT and 350 mM NaCl buffer onto the DEAE-FF column at a flow rate of 1 mL/min, ovocleidin-17 (OC-17) was obtained by loading 100 mM NaCl, 50 mM Tris, pH 8.0 on the CM-FF column at a flow rate of 0.5 mL/min. The purities of OCX-36, OC-17 and OC-116 were 96.82%, 80.15% and 73.22%, and the recovery rates were 55.27%, 53.38% and 36.34%, respectively. Antibacterial activity test suggested that phosphate solution extract exhibited significantly higher activity against the tested bacterial strains than the acetic acid or EDTA extract, probably due to more types of proteins in the extract. These results demonstrate that this separation method is feasible and efficient.
Keywords
eggshell matrix proteins; purification; ovocleidin-17; ovocalyxin-36; ovocleidin-116;
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