Background: Equine milk, including its whey proteins, is a source of nutrients and functional components in the human diet, and is especially beneficial for people with weakened immune systems, newborns, and athletes. Objectives Whey proteins in equine milk constitute approximately 20% of the total protein content and include various fractions such as albumin, globulin, and lactoferrin. Lactoferrin is one of the most extensively studied whey proteins in equine milk. Methods: HPLC-Mass analysis, enzymatic hydrolysis, modeling of 3D structure and biological activity in silico. Results: It has antioxidant, anti-inflammatory, and immunomodulatory properties, making it a promising candidate for influencing the various aspects of cardiovascular disease pathogenesis. The products of Lactoferrin hydrolysis by trypsin were confirmed using HPLC. The half-lives of the hydrolysate in the bloodstream and in an intestine-like environment were predicted in silico. Various biological activities (antihypertensive, anti-inflammatory, and antiangiogenic) were also estimated in silico and compared with the corresponding activities of lactoferrin hydrolysate amino acid sequences from camel and dromedary milk. Conclusions: The three-dimensional modeling of lactoferrin hydrolysate peptides was performed to support the development of computational models or simulations, as well as to investigate their potential antimicrobial, anti-inflammatory, or immune-modulating functions in clinical or nutritional applications.

The single chemical composition of mare milk, rich in whey proteins is similar to human milk. Lactoferrin is one of the important compounds contained in whey protein fraction, and has multiple biological functions such as antimicrobial and activation of human and animal immune system. Due to its strong antimicrobial activity, lactoferrin has potential pharmaceutical applications. The present work is mainly focused on the isolation and purification of the lactoferrin from mare’s milk. First, the lactoferrin from Kazakhstan mare milk has been purified by gel filtration Sephadex G-100 chromatography in two steps. The column of Sephadex G-100 was eluted with 0.01 M sodium phosphate buffer (pH 6.8). Lactoferrin enriched fractions were detected using UV absorbance at 280 nm and were identified in the first peak. Second, the purity of lactoferrin was checked by 12% SDS–PAGE and the molecular weight of lactoferrin (in the range of 80-82 kDa) was estimated using protein standard, which is recombinant human lactoferrin (expressed in rice, iron saturated, molecular weight 82.4 kDa).

Ethanol extracts of Hericium and Inonotus strains show strong antioxidant, antimicrobial activities and distinct phenolic profiles for functional applications.