Buttermilk, a potential material used to produce milk fat globule membrane (MFGM), is obtained as a byproduct of butter making from milk whole cream and cheese whey cream. This study investigated the effects of rennet and acid coagulation on the protein profiles of buttermilk rennet-coagulated whey (BRW) and buttermilk acid-coagulated whey (BAW). They were compared to those of whey cream buttermilk (WCB). Rennet coagulation was more efficient in removing casein, while retaining more IgG and lactoferrin than acid coagulation. BRW had more MFGM than BAW. Butyrophilin, xanthine dehydrogenase, and mucin1 were significantly higher (P < 0.05) in BRW, while fatty acid-binding protein 3 was enriched in BAW. KEGG analysis showed that complement and coagulation cascades had the greatest differences, and the abundance of proteins involved in this signaling pathway in BRW and BAW was higher, suggesting their potential anticoagulant and anti-inflammatory activity. BAW had higher apolipoprotein A4 and transcobalamin 2, which are essential carriers for transporting long-chain fatty acids and vitamin B12 from the intestine to the blood. Therefore, BAW intake might improve lipids and vitamin B12 absorption. This study can help deepen the understanding of protein composition of MFGM-enriched whey and facilitate the production of MFGM proteins for infants and old-aged populations.

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.

Dairy ingredients with highly concentrated protein contents are high added value products with expanding market. The manufacture of such ingredients includes a succession of unit operations of which heat treatment is a key step to guarantee the microbial safety, that induces major changes in protein structures and thus ingredients functionalities. However, due to an incomplete understanding of phenomena taking place at high protein concentrations, shedding light on their mechanisms is a scientific challenge as well as an industrial need. In this study, the influence of heat treatment (74 °C/ 30 s) of highly concentrated milk protein systems (up to 20 % w/w) on protein denaturation/aggregation and enzymatic coagulation properties was studied using an original semi-industrial approach. 10 % w/w protein solutions constituted with whey protein and casein micelles at milk ratio, standardized in osmosed water or ultrafiltration permeate were used. These protein solutions were processed in different ways prior the manufacture of powders: heat treatment of the 10 % w/w protein solution before vacuum evaporation, heat treatment of the 20 % w/w protein solution after vacuum concentration, two consecutive heat treatments before and after vacuum evaporation. A fourth powder was prepared from unheated 10 % w/w protein solution. An increase in protein concentration led to a higher heat-induced protein denaturation. This phenomenon was reduced when increasing the lactose content. The effect of heat treatment on the extent of protein denaturation was not cumulative. At high protein concentration, interactions between κ-casein and whey protein were modified compared to milk, as mainly micelle-bound aggregates were formed at pH about 6.7. This phenomenon was enhanced at low ionic strength and lactose content. Our study showed that the enzymatic coagulation properties of reconstituted protein powders could be correlated with their physico-chemical compositions. An increase in protein denaturation disrupted the gel reorganization and led to the formation of weaker gels but did not interfere on the micelles aggregation phase and the early gelation. On the contrary, an increase in ionic strength and lactose content led to higher gel time.

The objective of this work was to investigate the effect of pH adjustment (initial pH vs. pH 6.50) on the rennet-gelation properties of concentrates made by ultrafiltration (UF) and reverse osmosis (RO). Rennet-gelation kinetics were followed by dynamic rheology and κ-casein hydrolysis by reverse-phase HPLC. At initial pH, RO concentrates had better rennet-coagulation behavior than UF concentrates and skim milk, whereas adjusting the pH to 6.50 produced the opposite results. The kinetics of κ-casein hydrolysis were similar in skim milk, and both concentrates and were not affected by pH adjustment. Differences in rennet coagulation were then related to the extent of hydrolysis required to trigger casein micelle aggregation. Small pH adjustments (<0.2 pH unit) enabled the use of RO concentrate with similar rennet-gelation behavior to UF concentrate, despite major compositional differences. This study shows that pH adjustment of RO concentrates can be a simple approach to improve their coagulation properties; however, the mechanisms behind these improvements remain to be elucidated.

Pasteurized skim milk was subjected to (1) microfiltration (MF) at 50°C and (2) MF at 6°C after storage at 2°C. The products of these treatments were retentate (RMF50) and permeate (PMF50), and retentate (RMF6) and permeate (PMF6), respectively. Additionally, RMF50 was subjected to (3) cold MF after water dilution to produce retentate (RMF6R) and permeate (PMF6R). Calcium migration was monitored by analyzing ionic, soluble, and total calcium content in feed, retentates, and permeates. The influence of calcium partitioning and calcium addition to feed, retentates, and retentates diluted with water was determined. Without CaCl2 addition, only skim milk, RMF50, and RMF6 coagulated after rennet addition. Higher true protein and casein content of RMF50 and RMF6 resulted in shorter time of renneting. The retentates diluted with water showed no signs of coagulation within 40 min. The addition of PMF6R to RMF50 did not affect rennet coagulation time within the observed 40 min in comparison to RMF50 + water. In general, higher CaCl2 addition resulted in shorter rennet coagulation time. Special attention should be paid to calcium partitioning during membrane processing of cheesemilk. The level of calcium addition should be adopted to calcium content in such cheesemilk, which is affected by conditions of the filtration process (i.e., concentration factor and temperature).

BACKGROUND: Transglutaminase (TGase) modifies milk proteins by cross-linking of caseins, with increased cheese yield being the main technological benefit. In the present work the influence of TGase addition in different concentrations (0, 1, 2 and 3 U g(-1) protein in the system) and under different incubation conditions (0 h, 40 °C/2 h, 25 °C/4 h and 5 °C/16 h) on the rennet coagulation time (RCT) and the comprehensive rennet gel properties were investigated.
RESULTS: Modification of milk proteins by TGase in a concentration-dependent manner caused longer RCT and lower gel firmness. The highest TGase concentration and incubation at 40 °C for 2 h resulted in the longest RCT and the lowest gel firmness. Rennet gels obtained from TGase modified milk were characterised by significantly lower values of texture parameters, lower syneresis and were composed of smaller casein micelles, thinner chains and smaller clusters than those obtained from the control milk. The content of whey proteins in the gel from modified milk was higher and the content of individual casein fractions in the milk samples and rennet gels decreased upon TGase modification.
CONCLUSIONS: Rennet cheese with modified textural and nutritional properties and improved yield can be obtained upon TGase modification but simultaneous addition of rennet and TGase is recommended. © 2015 Society of Chemical Industry.

The effects of interaction of Asparagus racemosus (shatavari) with milk constituents and physico-chemical and functional characteristics of milk was studied. Addition of freeze dried aqueous shatavari extract at a concentration of 1 g /100 ml of milk showed a decrease in pH, rennet coagulation time and an increase in acidity, viscosity and heat stability at maximum. The extract also imparted brown colour to milk and showed an increase in a* (redness) and b* (yellowness) values but a decrease in L* (lightness) value. Proteins in milk were modified by reaction with shatavari extract. The derivatives formed were characterized in terms of SDS-PAGE. Electrophoretic pattern of sodium caseinate and whey containing 1% shatavari herb extract did not show any difference in band pattern i.e. there was no difference in mobility based on size of the proteins, but the intensity (width) of bands differed.