Biotechnological methods for separation of pigs pancreas glands protein substances with membrane technologies

The pancreas gland (PG) is a secondary product of livestock processing; it contains a wide range of biologically active compounds. The purpose of this article is to analyze the efficiency of technological approaches for pancreas gland extraction with the help of trehalose and a glycine-proline mixture aimed for recovery and separation of the gland's protein-peptide compounds. The extraction was conducted with 0.9% NaCl, 0.9% NaCl, with addition of 0.5 M trehalose (0.9% NaCl-0.5 M trehalose) and 0.9% NaCl with addition of 1% glycine and 0.1 M L-proline (0.9% NaCl-1% Gly-0.1M Pro), the ratio of pancreas gland to extractant was equal to 1:5. The concentration of the protein in the supernatants after their extraction was measured by the biuret reaction in a semi-automatic biochemical analyzer Biochem SA. The proteomic composition of the extracts and the native pancreas gland was assessed by one-dimensional Laemmli electrophoresis in a 12.5% polyacrylamide gel and by two-dimensional O'Farrell electrophoresis. When determining the intensity of the protein fractions, it was noted that the methodology of separation of protein-peptide mixtures extracted from the pigs pancreas gland with the extractant 0.9% NaCl-1% Gly-0.1M Pro, ensured the higher extraction of the proteins in comparison with the method of 0.9% NaCl-0.5 M trehalose. Notwithstanding the fact that application of amino acids (glycine and proline) mixture provided for a greater yield of proteins from the extract into the diafiltrate, the experiments in vitro showed that the diafiltrate obtained though trehalose featured higher activity. This may be explained by the fact that after dialysis removal of trehalose from the protein fraction with a molecular weight of less than 50 kDa, its residual quantities were still sufficient to prevent proteins aggregation and, as a consequence, the biological activity of the extracted proteins was preserved, while in the diafiltrate obtained through amino acids mixture where numerous protein aggregates were detected by 2-DE. This study allowed testing the biotechnological methodics on pig pancreatic tissues aimed to intensifying the extraction and separation of protein compounds. The results of the study are important for development of methodological approaches to obtaining the targeted substances for their further utilizing for various purposes.