Invited review: Modeling milk stability.

Novel insights into the stability of milk and milk products during storage and processing result from describing caseins near neutral pH as hydrophilic, intrinsically disordered, proteins. Casein solubility is strongly influenced by pH and multivalent ion binding. Solubility is high at a neutral pH or above, but decreases as the casein net charge approaches zero, allowing a condensed casein phase or gel to form, then increases at lower pH. Of particular importance for casein micelle stability near neutral pH is the proportion of free caseins in the micelle (i.e., caseins not bound directly to nanoclusters of calcium phosphate). Free caseins are more soluble and better able to act as molecular chaperones (to prevent casein and whey protein aggregation) than bound caseins. Some free caseins are highly phosphorylated and can also act as mineral chaperones to inhibit the growth of calcium phosphate phases and prevent mineralized deposits from forming on membranes or heat exchangers. Thus, casein micelle stability is reduced when free caseins bind to amyloid fibrils, destabilized whey proteins or calcium phosphate. The multivalent-binding model of the casein micelle quantitatively describes these and other factors affecting the stability of milk and milk protein products during manufacture and storage.

A quantitative calcium phosphate nanocluster model of the casein micelle: the average size, size distribution and surface properties.

Caseins (αS1, αS2, ß and κ) are the main protein fraction of bovine milk. Together with nanoclusters of amorphous calcium phosphate (CaP) and divalent cations, they combine to form a polydisperse distribution of particles called casein micelles. A casein micelle model is proposed which is consistent with the way in which intrinsically disordered proteins interact through predominantly polar, short, linear, motifs. Using the model, an expression is derived for the size distribution of casein micelles formed when caseins bind to the CaP nanoclusters and the complexes further associate with each other and the remaining mixture of free caseins. The result is a refined coat-core model in which the core is formed mainly by the nanocluster complexes and the coat is formed exclusively by the free caseins. Example calculations of the size distribution and surface composition of an average bovine milk are compared with experiment. The average size, size distribution and surface composition of the micelles is shown to depend on the affinity of the nanocluster complexes for each other in competition with their affinity for free caseins, and on the concentrations of free caseins, calcium ions and other salts in the continuous phase.

Size- and Dose-Dependent Body-Wide Organ Transcriptomic Responses to Calcium Phosphate Nanomaterials.

Nanomaterials are widely used in clinical practice. There are potential risks of body-wide infiltration due to their small size; however, the body-wide reliable risk assessment of nanoparticle infiltration is not fully studied and established. In this study, we demonstrated the size- and dose-dependent body-wide organ transcriptomic responses to calcium phosphate nanomaterials in vivo. In a mice model, a calcium phosphate nanocluster (amorphous calcium phosphate, ACP, ∼1 nm in diameter) and its crystallization product (ACP-M, ∼10 nm in diameter) in a series of doses was administrated systematically; multiorgan transcriptomics were then performed with tissues of heart, liver, spleen, lung, kidney, and brain to investigate the systematic effect of dose and size of nanomaterials on the whole body. The results presented gene expression trajectories correlated with the dose of the nanomaterials and tissue-specific risk effects in all detected tissues. For the dose-dependent tissue-specific risk effects, lung tissue exhibited the most significant risk signatures related to apoptosis, cell proliferation, and cell stress. The spleen showed the second most significant risk signatures associated with immune response and DNA damage. For the size-dependent tissue-specific risk effects, ACP nanomaterials could increase most of the tissue-specific risk effects of nanomaterials in multiple organs than larger calcium phosphate nanoparticles. Finally, we used the size- and dose-dependent body-wide organ transcriptomic responses/risks to nanomaterials as the standards and built up a risk prediction model to evaluate the risk of the local nanomaterials delivery. Thus, our findings could provide a size- and dose- dependent risk assessment scale of nanoparticles in the transcriptomic level. It could be useful for risk assessment of nanomaterials in the future.

Engineering artificial casein micelles for future food: Is casein phosphorylation necessary?

Industrial-scale production of recombinant proteins for food products may become economically feasible but correct post-translational modification of proteins by microbial expression systems remains a challenge. For efficient production of hybrid products from bovine casein and recombinant casein, it is therefore of interest to evaluate the necessity of casein post-translational phosphorylation for the preparation of hybrid casein micelles and study their rennet-induced coagulation. Our results show that dephosphorylated casein was hardly incorporated into artificial casein micelles but was capable of stabilising calcium phosphate nanoclusters with an increased size through adsorption on their surface. Thereby, dephosphorylated casein formed larger colloidal particles with a decreased hydration. Furthermore, the presence of increasing amounts of dephosphorylated casein resulted in increasingly poor rennet coagulation behaviour, where dephosphorylated casein disrupted the formation of a coherent gel network by native casein. These results emphasise that post-translational phosphorylation of casein is crucial for their assembly into micelles and thereby for the production of dairy products for which the casein micelle structure is a prerequisite, such as many cheese varieties and yoghurt. Therefore, phosphorylation of future recombinant casein is essential to allow its use in the production of animal-free dairy products.