Characterization of high pressure jet-induced fat-protein complexation

High-pressure-jet (HPJ) processing of various dairy systems has been shown to disrupt fat droplets and casein micelles and cause a strong association between fat and casein proteins. The present work seeks to better describe this association between fat and casein using a model milk formulated from confectionary coating fat (3.6% wt/wt), micellar casein (3.4% wt/wt), and water (93% wt/wt), which was then pasteurized, homogenized, and then either HPJ-treated (400 MPa) or not (non-HPJ-treated, control). Upon ultracentrifugation, fat in the non-HPJ-treated model milk creamed due to its low density. In the HPJ-treated model milk, fat precipitated with protein into a thick bottom layer upon ultracentrifugation, reflecting a strong association between protein and fat. Differential scanning calorimetry (DSC) and time-domain nuclear magnetic resonance of the non-HPJ-treated model milk revealed fat in 2 physical states: (1) fat that is physically similar to the bulk fat and (2) fat that was in smaller droplets (i.e., homogenized) and crystallized at a lower temperature than the bulk fat. In contrast, DSC of HPJ-treated model milks supported the presence of fat in 3 states: (1) fat that is physically similar to the bulk fat, (2) fat in small droplets that required substantial supercooling beyond the non-HPJ-treated model milk to crystallize, and (3) fat in such small domains that it crystallizes in a less stable polymorphic form than the non-HPJ-treated model milk (or does not crystallize at all). The state of fat within the HPJ-treated model milk changed minimally with acidification, indicating that the association is not dependent on the charge on the casein. Cryogenic transmission electron microscopy (Cryo-TEM) of the non-HPJ-treated model milk revealed uniform casein micelles, which likely adsorbed to the surface of fat globules post-homogenization. In contrast, Cryo-TEM of the HPJ-treated model milk revealed a porous protein aggregate that likely had dispersed fat throughout. Together, these results suggest that HPJ treatment causes fat to be entrapped by casein proteins in very small domains.