β-Casomorphin-7 (BCM-7) is a peptide released through the proteolysis of β-casein (β-CN), which is considered a bioactive peptide displaying evidence of promoting the binding and activation of the μ-opioid receptor located in various body parts, such as the gastrointestinal tract, the immune system and potentially the central nervous system. The possible effects of BCM-7 on health are a theme rising in popularity due to evidence found in several studies on the modulation of gastrointestinal proinflammatory responses that can trigger digestive symptoms, such as abdominal discomfort. With the advancement of studies, the hypothesis that there is a correlation of the possible effects of BCM-7 with the microbiota–gut–brain axis has been established. However, some studies have suggested the possibility that these adverse effects are restricted to a portion of the population, and the topic is controversial due to the small number of in vivo studies, which makes it difficult to obtain more conclusive results. In addition, a threshold of exposure to BCM-7 has not yet been established to clarify the potential of this peptide to trigger physiological responses at gastrointestinal and systemic levels. The proportion of the population that can be considered more susceptible to the effects of BCM-7 are evidenced in the literature review. The challenges of establishing the adverse effects of BCM-7 are discussed, including the importance of quantifying the BCM-7 release in the different β-CN genotypes. In summary, the reviewed literature provides plausible indications of the hypothesis of a relationship between β-CN A1/BCM-7 and adverse health effects; however, there is need for further, especially in vivo studies, to better understand and confirm the physiological effects of this peptide.

The structure, stability, and hydrolysis characteristics of beta-lactoglobulin (LG) A are different from those of either beta-LG B or beta-LG C. Purified samples of these proteins were hydrolyzed with trypsin and the rates of loss of native monomeric beta-LG structure were measured using sodium dodecyl sulfate PAGE. At the same time, the appearance of many individual peptides were identified and followed in time by HPLC, measuring their concentration as a function of solution pH, temperature, protein concentration, and added urea or palmitate. The identity of the peptides was confirmed by liquid chromatography-mass spectrometry. This semiquantitative exploration showed that the rate of hydrolysis was in the order beta-LG A > beta-LG B > beta-LG C under most circumstances, and that 12 of the 18 trypsin-susceptible bonds were cleaved at very similar rates that were governed by the variant type. Consequently, the rate of hydrolysis of the intact protein was related to the overall structural stability of the individual proteins and the accessibility of certain peptide bonds to the enzyme. The hydrolysis of mixtures of 2 or more variants or of denatured beta-LG gave more heterogeneous peptide mixtures.

Whey-protein-free skim milk samples were prepared by a combination of ultrafiltration and microfiltration techniques. The desired concentrations of alpha -lactalbumin and the A, B or C variant of beta -lactoglobulin (beta -LG) were added back to the milk. The milk samples were heated at temperatures from 75 to 100 degreesC and the rates of beta -LG denaturation were determined using quantitative polyacrylamide gel electrophoresis. At all temperatures, the rate of denaturation of beta -LG was most rapid for the B variant, with the A variant intermediate and the C variant most thermostable. The differences in thermostabilities were quite large at temperatures below about 85 degreesC, but much smaller at higher temperatures. The differences in thermostabilities between the different variants of beta -LG are probably related to the differences in structure, charge and self-association behaviour that are induced by the small changes in primary sequence between the variants.

The effect of fortification of reconstituted skim milk with different levels of a whey protein mixture containing a 1:2 ratio of α-lactalbumin (α-la) and different genetic variants of β-lactoglobulin (β-LG) on the rheological properties of acid milk gels, formed by acidification with glucono-δ-lactone, was investigated. Milk samples were either unheated or heated at 80°C for 30 min before acidification. Acid gels prepared from unheated skim milk had very low G′ values, long gelation times and low gelation pH. Samples prepared from heated milk had markedly higher G′ values, a reduced gelation time and an increased gelation pH. The addition of increasing levels of whey protein mixtures containing β-LG B or β-LG C to the milk prior to heating and acidification caused an almost linear increase in the G′. In contrast, whey protein mixtures containing β-LG A caused a progressive increase in the G′ with added protein levels up to about 0.7% (w/w) but little further change at higher addition levels. A mixture of the A and B variants of β-LG gave an intermediate behaviour between those of the A and B variants. In all samples, the G′ value at 5°C was approximately twice that at 30°C so that the relative differences as a result of the β-LG genetic variants were similar for the two temperatures.

The combined effect of genetic variations of β-lactoglobulin and κ-casein on the heat stability of milk was studied. The phenotype combination β-lactoglobulin AA with κ-casein BB was associated with the highest maximum coagulation time.

The different rates of cleavage by chymosin of κ-casein variants has been studied. Differences between the C variant and the A and B variants are attributed to the direct effect of the Arg to His substitution at position 97 in the κ-casein sequence.

The emulsifying properties of three genetic variants of β-lactoglobulin (β-lac) (the A, B and C variants) are investigated as a function of protein concentration. Differences in the emulsifying properties and emulsion stability are explained in view of the known differences in physico-chemical and structural/conformational properties of the β-lac variants. β-lac A forms the finest emulsion droplets, and β-lac C the largest droplets. The order of decreasing emulsifying ability (A>B>C) can be explained in terms of differences in the molecular structure, and conformational stability of the variant proteins. The creaming stability, when compared at the same particle size, is greatest for β-lac C, with β-lac A and B having a similar and lower stability. The differences in creaming stability may arise from a higher surface coverage for the β-lac C droplets at an equivalent particle size. The storage stability is lower for β-lac A than for β-lac B and C, which both show a similar behaviour. Storage stability differences are discussed in terms of differences in molecular structure, conformational stability, interfacial viscosity and surface coverage for the three variants.

The objective of this study was to investigate the effects of contrasting nutritional regimens on milk composition from cows of different protein phenotypes. Twenty sets of seasonally calving identical twin cows that constituted five different protein phenotypes (four sets of twins per phenotype) were subjected to two nutritional treatments in crossover experiments during spring (early lactation) and summer (mid to late lactation). The phenotypes studied allowed a comparison of the AA, AB, and BB variants of both beta-lactoglobulin (beta-LG) and kappa-casein. Nutritional treatments were 1) ad libitum grazing (i.e., cows were allocated a pasture allowance of approximately 40 kg of dry matter/d per cow) plus 5 kg of a concentrate based on barley and 2) restricted grazing (pasture allowance of 20 kg of dry matter/d per cow). Milk samples were collected from each cow near the end of each 14-d treatment period and were analyzed for a detailed range of individual protein and fat constituents. Diet had significant effects on the concentrations of all milk components measured. Protein phenotype affected some protein components but not fat components. Interactions between the effects of beta-LG phenotype and diet were noted for the concentrations of some milk components. Diet and protein phenotype have important effects on the manufacturing potential of milk produced under the dairying systems of New Zealand, which rely heavily on grazing. The effects of nutrition on milk composition may depend on the beta-LG phenotype.

The rheological properties of heat-induced gels made from beta-lactoglobulin variants A, B and C were compared. The relative G ' values (elastic moduli) for gels formed in 90 mM NaCI solutions were A = B > C. Conversely, in 30 mM CaCl2 the relative G ' values were C > A = B. The differences in theological properties were due to A and B variants forming less rigid gels in CaCl2 (similar to 7 kPa) than NaCI (similar to 20 kPa), and variant C forming gels of similar rigidity in both salt solutions. It was concluded that genetic variation in P-lactoglobulin changes the effect of salts on gelation but does not cause a universal increase or decrease in gel forming ability.

The action of plasmin and chymosin on genetic variants of αs1-casein has been studied. Little difference was found between the action of plasmin on αs1-casein, A, B and C. However, chymosin hydrolysed αs1-casein A more slowly than the B and C variants.

Two experiments (mid- and late lactation) were conducted to test whether methionine (Met) limited milk production in Friesian cows fed sole diets of fresh perennial ryegrass–white clover pasture. In mid-lactation (exp. 1), 26 cows were assigned to one of three treatments: ruminally protected oral DL-Met (15 g d−1), continuous intrajugular infusion of L-Met (15 g d–1) and control. Twelve animals from exp. 1 were used during late lactation (exp. 2) and distributed in two experimental groups: ruminally protected oral DL-Met (15 g d−1) and control. Dry matter, metabolizable energy, crude protein and Met intakes, milk yield and composition and blood Met, cysteine and urea were measured. Oral and intravenous Met supplementation increased blood Met concentration by 50–90% compared with controls. Met supplementation did not alter the concentration of milk fat, protein or lactose in either experiment. Met supplementation had no significant effects on yields of fat, lactose, casein, whey proteins or non-protein nitrogen during mid-lactation. In late lactation, Met supplementation did not affect milk protein composition or yield of milk components, with exception of a decrease (P < 0.05) in the yield of β -casein. Intravenous Met supplementation increased (P < 0.05) the efficiency of conversion of pasture N to milk protein in mid-lactation. Key words: Dairy cows, methionine, ruminally protected methionine, milk protein, casein, fresh pasture diets

The stability of the casein micelle is dependent on the presence of kappa-casein (CN) on the surface of the micelle where it functions as an interface between the hydrophobic caseins of the micelle interior and the aqueous environment. kappa-Casein is also involved in thiol-catalyzed disulfide interchange reactions with the whey proteins during heat treatments and, after rennet cleavage, in the facilitation of micelle coagulation. These functions of kappa-CN are regulated by the three-dimensional structure of the protein on the micelle surface. The usual means of determining structure are not available for kappa-CN because this protein is strongly self-associating and has never been crystallized. Instead, algorithms were used to predict selected secondary structures and circular dichroism spectroscopy on kappa-CN and the macropeptide released by chymosin. Three peptides were synthesized to cover the chymosin-sensitive site (His98-Lys111), the region in the macropeptide that could be helical (Pro130-Ile153), and the region between. Nuclear magnetic resonance spectroscopy showed that the peptide His98-Lys111 was probably a beta-strand with tight turns at each end. This hypothesis was confirmed by a study of the molecular dynamics showing that the C variant of kappa-CN interacted less strongly with chymosin; consequently, the slow renneting time of milk that contains this protein was explainable. Both circular dichroism and nuclear magnetic resonance indicated that the peptide Pro130-Ile153 was probably helical under normal physiological conditions. A preliminary study using nuclear magnetic resonance showed that the intervening peptide had no discernible secondary structure. Consequently, most of the beta-sheet structure of kappa-CN is likely in the para-kappa-CN region.