Storozh L. Biotechnology of functional phosphopeptides from proteins of casein complex of cow`s milk

Іn the dissertation work the technology of production of biologically active phosphopeptides from the casein complex of cow's milk is scientifically substantiated and developed. As a substrate, a number of phosphoproteins from cow's milk have been isolat-ed in conditions that minimally affect their structure and chemical composition. Two prep-arations of total casein (isoelectric deposition and deposition in the system «water – milk proteins – pectin») were received. By means of exclusive chromatography it was isolated phosphoroprotein of β-CN-5P and by the differential precipitate in combination with a specific proteolysis with recombinant chymosin it was obtained a mixture of phosphoproteins, which included αS1-СN- ХР and αS2-СN-ХР fractions. For substationation the method of proteolytic cleavage of isolated phosphoprotein precursors were used proteolytic preparations: papain, trypsin, chymotrypsin, pancreatin, neutral protease. The obtained results testify to the expediency of using as a substrate of the general preparation of milk phosphoproteins and of the proteolytic preparation of pancreatin to produce natural bioactive phosphopeptides. The influence of the enzyme-substrate ratio, pH and temperature on the yield of phosphopeptides was investigated. As a result, conditions were provided that ensure a high yield of phosphopeptides, as well as the formation of natural biologically active phosphopeptides (pH 7.9; 37°C; the ratio of «en-zyme: substrate» – from 1:100 to 1:140). On the basis of the conducted research the tech-nological scheme of production of biologically active phosphopeptides has been devel-oped, its testing in industrial conditions has been carried out. The resulting preparation of bioactive phosphopeptides is characterized by molecular weight distribution. It has been established that more than 52% of phosphopeptides have a molecular weight of 700-1500 Da. It has been experimentally found that isolated phosphopeptides can inhibit the growth of cancer cells: human glioblastoma U373 and human adenocarcinoma HeLa.