A Study of Various Chemical Pre-treatments to Fractionate Lipids from Whey Protein Phospholipid Concentrate
Date
2021Author
Swaminathan, Aakash Varsha
Molitor, Mike S
Burrington, Kimberlee J
Otter, Don
Lucey, John A
Publisher
Journal of Dairy Science
Metadata
Show full item recordAbstract
Dairy-derived lipids such as phospholipids (PL) have been gaining interest due to their functional and nutritional properties. Our research goal was to develop a separation process (non-solvent based) to produce an enriched dairy lipid fraction from whey protein phospholipid concentrate (WPPC). Various chemical pre-treatments (i.e., adjustment of pH, calcium, or temperature) were applied to rehydrated commercial WPPC solutions. These treatments were done on a bench top scale to aid in the precipitation of proteins or PL. The chemically treated solutions were centrifuged and fractionated into three layers: (1) top fat layer, (2) supernatant in the middle zone, and (3) sediment at the bottom of the centrifuge tubes. The thickness/size of the layers varied with the treatment parameters. Compositional analysis of each layer showed that the proteins, fat, and PL always appeared to fractionate in similar proportions. The proteins in each layer were characterized using SDS-PAGE under reducing and non-reducing conditions. Different proteins including whey proteins, caseins and milk fat globule membrane proteins/lipoproteins were identified and no specific type of protein had an affinity for either the top or bottom layer. All types of proteins were present in each of the layers after centrifugation and there were no major differences in fractionation of the proteins between layers with respect to the chemical treatment applied. The microstructure of protein and fat in WPPC was investigated using confocal laser scanning microscopy (CLSM). Dual staining of the rehydrated WPPC solution with Fast Green FCF (proteins) and Nile Red (lipids) showed the presence of very large protein aggregates that varied in size from 20 to 150 μm, with fat trapped within these aggregates. CLSM images of liquid WPPC revealed fine strands of a weak protein network surrounding the fat globules. This indicated that there were specific interactions between the proteins, as well as between the fat and proteins in WPPC. SDS treatment was performed to understand the nature of the interactions between protein and fat. It was found that about 35% of the fat present in WPPC was in the form of free fat, which was only physically entrapped within the protein aggregates. The remaining fat has some form of association with the proteins in WPPC. Other fractionation techniques would be needed to obtain an enriched dairy lipid fraction.
Subject
Whey protein phospholipid concentrate; chemical pre-treatments; phospholipids
Permanent Link
http://digital.library.wisc.edu/1793/82259Type
Working Paper