The Recovery of Protein from Egg Yolk Protein Extraction Granule Byproduct

Abstract

In addition to proving an excellent source of nutrients, eggs are used in the food, cosmetic, and biotechnology industries for their rheological and bioactive properties. Much of the potential for the added value is in individual components of the egg, rather than the whole egg. At low speed centrifugation, yolk separates into two distinct fractions-granules and plasma. It is becoming increasingly popular in the industry to remove the plasma fraction of the egg yolk to use for its livetins, particularly immunoglobulin Y, leaving behind a granule by-product ("yellow cake"). Previous research has shown potential added-value from the granule fraction, especially from its phosvitin and phospholipids. Granules are protein aggregates with complexes of phosvitin and high density lipoproteins linked by phosphocalcic bridges. In their native form, the proteins are mostly insoluble, however previous studies have shown the links can be broken by alterations in pH, ionic strength, and mechanical treatments. This thesis project seeks to find potential uses for the egg yolk by product after the removal of the livetin fraction by means of further fractionation with mechanical treatment (filtration). Two variables were tested to extract more proteins from the yellow cake. Salt was added to 10% solids solution of yellow cake in water before filtration at four different NaCl levels: 0%, .05%, 1%, and 2.5%. Additionally pH was tested at four different levels: 4.6, 4.8, 5.0, 5.2. The samples were also tested for antibacterial properties against Escherichia v coli with a minimum inhibitory concentration assay (MIC). Analysis with BCA showed salt concentration had a significant effect on the yield of protein. The highest concentration of salt tested, 2.5%, had the highest protein yield. Additionally, SDS-PAGE showed 2.5% salt had the most unique protein bands. This could be to the disruption of the phosphocalcic links between the phosvitin and HDL by NaCl, allowing the protein to solubilize. pH did not have a significant effect on the yield or types of proteins in the range tested in this experiment. There is no conclusive evidence of antibacterial properties against E. coli from the protein extract. The MIC assay had growth show up in all wells with the protein extract, however there was a visible decrease in turbidity with higher concentration of the protein extract. This could mean that the protein extract does have some antibacterial properties, but needs testing at higher concentrations or with isolated proteins/peptides. The SDS-PAGE revealed bands showing phosvitin present, which has known antibacterial properties. Overall, improvements to the methods for further protein extraction from egg yolk by-products will help lead the industry to finding novel uses and product applications. vi ACKNOWLEDGMENTS I would like to thank my advisor, Dr. Rafael Jimenez-Flores, for his guidance and support, all the way from San Luis Obispo, CA to Indianapolis, IN to Columbus, OH.