The main goal of this study was to assess the potential proteins of goat milk (i.e. α‐s1‐casein, α‐s2‐casein, β‐casein, κ‐casein, α‐lactoglobulin and β‐lactalbumin) as precursors of antimicrobial peptides (AMPs). Bioinformatics tools such as BIOPEP‐UWM (enzyme action) were used for the in silico gastrointestinal digestion via a cocktail of pepsin, trypsin, and chymotrypsin A. The antimicrobial activity of peptides was predicted by using four algorithms, including Random Forest, Support Vector Machines, Artificial Neural Network and Discriminant Analysis on CAMPR3 online server, which works on Hidden Markov Models. Different online tools predicted the physiochemical properties, allergenicity, and toxicity of peptides as well. In silico gastrointestinal digestion simulation of proteins by enzymes cocktail yielded a total of 83 potential AMPs, with thirteen peptides being confident by all four algorithms. More AMPs were released from β‐casein (21) than from β‐lactoglobulin (16), α‐s1‐casein (15), α‐s2‐casein (12), κ‐casein (11) and α‐lactalbumin (9). A total of 17 peptides were cationic, and the majority of the peptides were extended AMPs. These peptides were released from α‐s1‐casein (SGK, IQK), α‐s2‐casein (SIR, AIH, TQPK), β‐casein (GPVR, AVPQR, AIAR, GVPK, SQPK, PVPQK, IH, VPK), k‐casein (AIPPK, QQR, IAK, TVPAK). All of the AMPs were anticipated to be non‐toxic, and 54 of the 83 peptides were confirmed to be non‐allergic, with the remaining 29 suspected of being allergenic and 31 to be predicted to have good water solubility. Further the molecular docking was used to evaluate the potent dihydropteroate synthase (DHPS) inhibitors. On the basis of ligand binding energy, 17 predicted AMPs were selected and then analyzed by AutoDock tools. Among the 17 AMPs, 3 AMPs were predicted as high‐potent antimicrobial. Based on these findings, in silico investigations reveal that proteins of goat milk are a potential source of AMPs. These peptides can be synthesized and improved for use in the food sector. Practical applications Goat milk is regarded as a high‐quality milk protein source. According to this study, goat milk protein is a possible source of AMPs, and therefore, most important AMPs can be synthesized and developed for use in the food sector.

An in silico approach was used for hydrolysis of sheep milk proteins (α‐s1, α‐s2, β‐casein, κ‐Cn, α‐lactalbumin, and β‐lactoglobulin) by gastrointestinal enzymes in order to generate bioactive peptides (BAPs) that can inhibit ACE and DPP‐IV. Sheep milk proteins showed higher similarity with goat milk proteins. These data were acquired via the Clustal Omega tool to perform sequence alignment analysis. The BIOPEP‐UWM database was used to examine the ability of sheep milk protein sequences to generate BAPs, which included a description of their potential bioactivity as well as the frequency of fragments with specified activities. Using the “Enzyme(s) action” tool (BIOPEP‐UWM), digestive enzymes pepsin, trypsin, and chymotrypsin, and three enzyme combinations were selected to computationally hydrolyze milk proteins for obtaining information about ACE and DPP‐IV inhibitory peptides. Other online programs were used to test potential peptides for bioactivity, toxicity, and physicochemical properties. BAPs produced from PTC‐hydrolyzed proteins were analyzed using a peptide ranker, and their inhibitory effects on ACE and DPP‐IV were determined using molecular docking. Consequently, the results of molecular docking analysis show that the peptide PSGAW (αS1‐Cn f155–159) binds to DPP‐IV with binding energy (−8.9 kcal/mol). But in the case of ACE, two potential BAPs were selected: QPPQPL (β‐Cn f161‐166) and PSGAW. These two BAPs revealed a higher binding affinity for ACE with a binding energy of −9.8 kcal/mol. Thus, the results showed that sheep milk proteins were a promising source of antidiabetic and hypotensive peptides. However, experimental and pre‐clinical studies are necessary to assay their therapeutic effects. Practical applications Sheep milk proteins are known as a high‐quality milk protein resource. Effective enzymatic hydrolysis of sheep milk proteins can release bioactive peptides and also release potential ACE and DPP‐IV inhibitory peptides. This in silico study specifies a theoretical root for sheep milk proteins as a novel source of potential bioactive peptides and may offer guidance for invitro hydrolysis of proteins for the production of bioactive peptides valuable for human consumption.

Antimicrobial peptides (AMPs) are emerging as promising novel drug applicants. In the present study, goat milk was fermented using Lactobacillus rhamnosus C25 to generate bioactive peptides (BAPs). The peptide fractions generated were separated using ultrafiltration membranes with molecular weight cut‐offs of 3, 5, and 10 kDa, and their antimicrobial activity toward Gram‐positive and Gram‐negative bacteria was investigated. Isolated AMPs were characterized using RP‐HPLC and identified by LC–MS/MS. A total of 569 sequences of peptides were identified by mass spectrometry. Out of the 569, 36 were predicted as AMPs, 21 were predicted as cationic, and out of 21, 6 AMPs were helical peptides. In silico analysis indicated that the majority of peptides were antimicrobial and cationic in nature, an important factor for peptide interaction with the negative charge membrane of bacteria. The results showed that the peptides of <5 kDa exhibited maximum antibacterial activity against E. faecalis, E. coli, and S. typhi. Further, molecular docking was used to evaluate the potent MurD ligase inhibitors. On the basis of ligand binding energy, six predicted AMPs were selected and then analyzed by AutoDock tools. Among the six AMPs, peptides IGHFKLIFSLLRV (−7.5 kcal/mol) and KSFCPAPVAPPPPT (−7.6 kcal/mol), were predicted as a high‐potent antimicrobial. Based on these findings, in silico investigations reveal that proteins of goat milk are a potential source of AMPs. This is for the first time that the antimicrobial peptides produced by Lactobacillus rhamnosus (C25) fermentation of goat milk have been identified via LC–MS/MS and predicted as AMPs, cationic charges, helical structure in nature, and potent MurD ligase inhibitors. These peptides can be synthesized and improved for use as antimicrobial agents. Practical applications Goat milk is considered a high‐quality source of milk protein. According to this study, goat milk protein is a potential source of AMPs, Fermentation can yield goat milk‐derived peptides with a broad antibacterial activity spectrum at a low cost. The approach described here could be beneficial in that the significant AMPs can be synthesized and used in the pharmaceutical and food industries.

Goat milk and protein hydrolysate peptides have generated significant interest due to their diverse range of biological functions. To produce these peptides, casein (CP), and whey proteins (WP) were partially purified from goat milk and subsequently hydrolyzed with gastrointestinal endopeptidases such as pepsin, trypsin, and α‐chymotrypsin based on in silico selection to achieve gastrointestinal stable peptides. The hydrolysates were characterized using various techniques, including zeta sizer, polydispersity index (PDI), zeta potential, and reverse‐phase high‐performance liquid chromatography (RP‐HPLC). The resulting peptides were then purified using RP‐HPLC. To assess cytotoxicity, an MTT (3‐[4, 5‐dimethylthiazol‐2‐yl]‐2, 5‐diphenyl tetrazolium bromide) assay was performed with fluorescence microscopy. Finally, cytokine levels were determined using enzyme‐linked immunosorbent assay (ELISA). In comparison to WP and CP, the casein protein hydrolysates (CPH) and whey protein hydrolysates (WPH) peptides average particle size and PDI were decreased. The zeta potential of casein and whey peptides hydrolysates there were no differences. The RP‐HPLC analysis revealed the production of peptides through the hydrolysis of CP and WP. These hydrolysates peptides were found to possess diverse biological activities, as evidenced by their significant inhibition of α‐amylase, pancreatic lipase (PL), and angiotensin‐converting enzyme (ACE), as well as their antibacterial properties against selected pathogens. Further investigations were carried out to understand the mode of action of these peptides using fluorescence and scanning electron microscope (SEM) microscopy. This study provides the first evidence of gastrointestinal stable peptides derived from goat milk with inhibitory activity against α‐amylase, PL, and ACE, as well as antibacterial properties against specific pathogens. Additionally, the peptides demonstrated significant cytokine inhibition and low toxicity toward the HT‐29 cell line. This study strongly suggests that the biologically active peptides responsible for the health‐related bioactive properties described here, which are considered functional and nutraceutical ingredients, should be identified and validated in subsequent research, particularly the most effective hydrolysate peptides derived from goat milk protein. Goat milk is widely recognized as an excellent source of milk protein. In this study, bioactive peptides were produced using casein and whey proteins from goat milk, which exhibited inhibitory activity against angiotensin converting enzyme, α‐amylase, and pancreatic lipase, as well as antibacterial properties. Goat milk protein is a promising source for developing high‐quality protein products with excellent safety standards that have potential applications in the pharmaceutical and food industries.

The medicinal plants have been used by humans since ancient times, and the great civilizations of the world in ancient times were well aware of the benefits brought by the use of medicinal plants. This chapter provides important information regarding medicinal plants that have a wide variety of antioxidative agents ranging from bitter compounds that stimulate digestion system, phenolic compounds for antioxidant and numerous other pharmacological properties, antibacterial, and antifungal to tannins that act as natural antibiotics, diuretic substances, alkaloids, and so forth.