Disruption and sedimentation of casein micelles and casein micelle isolates under high-pressure homogenization

Native casein micelles were isolated from raw skim milk by ultrafiltration (< 30 kDa) or microfiltration (< 0.2 μm) and subjected to high-pressure homogenization (HPH) at 100, 200, 250, 300, and 350 MPa. Of particular interest was the effect of HPH on casein micelle size in solutions varying in ionic strength (0, 5, 10, and 15 mM CaCl₂) and micelle size populations. Particle size distribution reflected an initial decrease in micelle diameter in all samples at 100 MPa. In samples containing 10 and 15 mM CaCl₂, there was an abrupt increase in particle size and subsequent casein precipitation followed by sedimentation upon centrifugation at elevated pressures (300 and 350 MPa). The amount of sedimentable casein protein increased as CaCl₂ concentration (10 and 15 mM) and pressure (300 and 350 MPa) increased as determined by UV absorbance of sample supernatant. SDS-PAGE indicated extensive micellar disruption at elevated pressures (300 and 350 MPa) and confirmed that the sedimented portion of the samples contained casein proteins and minimal amounts of whey proteins. Results indicated that through HPH treatment casein micelle size can be modified based on CaCl₂ concentration and pressure applied. Based on these findings, HPH in combination with an appropriate suspending medium has the ability to modify micelles to a desired size for a number of potential applications. Industrial relevance: The modification of structure-function properties of the casein micelle from bovine milk by using high-pressure homogenization is relevant in (1) the development of new ingredients to change rheological/textural properties of dairy based foods, and (2) the discovery of new and/or improved functionalities for protein quaternary structures.