Hydrolysis of ultra-filtrated retentate of whey can be an energy- and cost-effective method for the direct production of WPH from whey compared to the industrial production of WPH from whey protein concentrate. This study suggests that WPH with good nutritional and biological properties can be effectively used in health-promoting foods as a biofunctional ingredient.

ACE inhibitory and antioxidative peptides identified by LCMS/MS, from mixed milk (Bubalus bubalis and Bos taurus) tryptic whey protein hydrolysate, were compared with the in silico predictions. α la and ß lg sequences, both from Bubalus bubalis and Bos taurus, were used for in silico study. SWISS-PROT and BIOPEP protein libraries were accessed for prediction of peptide generation. Study observed gaps in the prediction versus actual results, which remain unaddressed in the literature. Many peptides obtained in vitro, were not reflected in in silico predictions. Differences in identified peptides in separate libraries were observed too. In in silico prediction, peptides with known biological activities were also not reflected. Predictions, towards generation of bioactive peptides, based upon in silico release of proteins and amino acid sequences from different sources and thereupon validation in relation to actual results has often been reported in research literature. Given that computer aided simulation for prediction purposes is an effective research direction, regular updating of protein libraries and an effectual integration, for more precise results, is critical. The gaps addressed between these two techniques of research, have not found any address in literature. Inclusion of more flexibility with the variables, within the tools being used for prediction, and a hierarchy based database with search options for various peptides, will further enhance the scope and strength of research.

Caseinophosphopeptides (CPPs) are multifunctional bioactive peptides containing phosphorylated seryl residues in their sequence. In the present study, method for the production of CPPs from buffalo milk casein was optimised and characterised for their sequence, calcium solubilising and calcium binding activities. Response surface methodology was used to optimise the conditions for hydrolysis of buffalo casein by trypsin to obtain maximum yield of CPPs. The optimum hydrolysis conditions were as follows: hydrolysis pH 7.5, temperature 37 °C, hydrolysis time 7.0 h. Under these conditions, the experimental yield obtained was 10.04±0.24%, which is slightly lower than value predicted by the model. These CPPs were able to solubilise 1.03±0.08 mg la/mg CPPs in presence of excess phosphate and bind 0.935 mg of Ca/mg of CPPs. Eight phosphopeptides, i.e., α(s1)-CN f (37-58) 2P; α(s1)-CN f (37-58) 3P; α(s1)-CN f (35-58) 2P; α(s1)-CN f (35-58) 3P; α(s2)-CN f (2-21) 4P; α(s2)-CN f (138-149) 1P; β-CN f (2-28) 4P and β-CN f (33-48) 1P were identified by LC-MS/MS which contained motif for binding of divalent minerals. The sequences of these CPPs differed from that of derived from bovine casein.

The accelerated emergence of antimicrobial resistance (AMR) among diverse pathogens poses a global public health threat. Biofilm-related infections of epithelial surfaces, wounds, implants, or medical devices that are non-responsive to standard antibiotics require the use of higher-order antimicrobials leading to toxic side effects and microbiome dysbiosis. Owing to the multifarious beneficial effect and inherent safety, probiotic therapy is emerging as a preferred approach for preventing and treating not only enteric infections but also as an alternative to antibiotic therapy against a plethora of diseases. This study reports the isolation of novel gut probiotic bacteria from the small ruminant livestock- domestic goat (Capra hircus) with antimicrobial and antibiofilm activity against the 'ESKAPE' group of pathogens, and comprehensive microbiological, biochemical, and 16S-rRNA gene V1-V3 region, and 16S-23S ISR region based molecular characterization of 20 caprine gut-derived Lactobacillus spp. (LAB). Of these, six selected LAB isolates exhibited considerable biofilm formation in anaerobic conditions and broad-spectrum growth inhibitory and anti-biofilm activity against 'ESKAPE' pathogens. Further, these LAB isolates were also found to be susceptible to the European Food Safety Authority (EFSA) indexed antibiotics within the cut-off Minimum Inhibitory Concentration (MIC) limits, indicating their potential safety as feed additives. Altogether, this study demonstrates the beneficial probiotic characteristics of caprine gut-derived LAB isolates and their anti-bacterial activity, substantiating their potential usage as an alternative to antibiotics or as direct-fed microbial (DFM) feed supplements in livestock farming.