The experiments reported in this research paper examine the potential of digestion using acidic enzymes Protease A and Protease M to selectively hydrolyse α-lactalbumin (α-La) whilst leaving β-lactoglobulin (β-Lg) relatively intact. Both enzymes were compared with pepsin hydrolysis since its selectivity to different whey proteins is known. Analysis of the hydrolysis environment showed that the pH and temperature play a significant role in determining the best conditions for achievement of hydrolysis, irrespective of which enzyme was used. Whey protein isolate (WPI) was hydrolysed using pepsin, Acid Protease A and Protease M by randomized hydrolysis conditions. Reversed-phase high performance liquid chromatography was used to analyse residual proteins. Regarding enzyme selectivity under various milieu conditions, all three enzymes showed similarities in the reaction progress and their potential for β-Lg isolation.

The aim of this work was to evaluate the impact of two factors on the ripening profiles of hard cooked cheeses: (F1) the growth medium for the primary and adjunct cultures, constituted by autochthonous strains: Lactobacillus helveticus 209 (Lh209) and Lactobacillus paracasei 90 (Lp90), respectively, and (F2) the addition of L. paracasei Lp90 as adjunct culture. Four types of cheeses were made: W and M cheeses in which only Lh209 was added after its growth in whey and MRS, respectively; Wa and Ma cheeses in which both strains (Lh209 and Lp90) were added after their growth in whey and MRS, respectively. Physicochemical and microbial composition, proteolysis and profiles of organic acids and volatile compounds were analyzed. According to the methodology of the cultures preparation, W and Wa cheeses showed a higher level of secondary proteolysis and lower level of primary proteolysis (P < 0·05), lower content of citric and acetic acids and higher amount of propionic acid (P < 0·05), in comparison with M and Ma cheeses. The incorporation of Lp90 increased the secondary proteolysis (P < 0·05), decreased the citric acid (P < 0·05), and increased the propionic acid only when was added after their growth in whey (P < 0·05). Both factors significantly modified the percentages of the volatile compounds grouped in chemical families; in addition, for the half of the compounds detected, significant differences were found. Based on the obtained results, the use of Lp90 as an adjunct in hard cooked cheeses, and the preincubation of the cultures in whey are strategies to accelerate the cheese ripening and to enhance the production of some characteristic compounds of this type of cheeses, such as propan-2-one, hexan-2-one, 2- and 3-methyl butanal, heptan-2-ol, acetic and 3-methylbutanoic acids and 3-hydroxy butan-2-one.

We evaluated the effect of in vitro digested milk on mature adipocytes 3T3-L1, paying particular attention to its fatty acid composition, and comparing human (HM), donkey (DM), bovine (BM), ovine (OM), caprine (CM) and formula (FM) milk. Cellular viability, apoptosis, oxidative response and gene expression levels of NF-κB p65, HMGB1, SREBP-1c and FAS were evaluated. Digested milk treatments significantly reduced 3T3-L1 mature adipocytes viability and caspase activity compared with control group, but no significant differences were observed among different sources of digested milk. In all digested milk samples, ROS level was higher than the control, however, the digested human and formula milk showed lower levels of ROS than DM, BM, OM and CM samples. Lower capacity of HM and FM to induce oxidative stress in mature adipocytes was ascribed to the peculiar free fatty acids profile of digested milk samples. All milk treatments elicited a significant over-expression of NF-κB p65 in 3T3-L1 adipocytes compared to the control; the lowest gene expression was found in HM, BM, OM and CM, the highest in FM and an intermediate behavior was shown in DM. All digested milk treatments influenced the gene expression of SRBP-1c with FM and HM showing the highest levels. For FAS expression, BM showed the highest level, OM and CM intermediate and FM, HM and DM the lowest levels, however HM and DM had comparable levels to the control.