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Vol. 25 (2022 year), No. 3, DOI: 10.21443/1560-9278-2022-25-3
Tikhonov S. L., Tikhonova N. V., Kolberg N. A., Ozhgikhina A. S., Shikhalev S. V.
Microcapsulation and evaluation of subchronic toxicity of peptides extracted from cow colostrum and peptides of Fabricius bursa extract of broiler chickens
Peptides have high biological activity and are used in the composition of medicines. Peptides are not widely used in food technology as they are sensitive to the action of proteolytic enzymes of the gastrointestinal tract and are characterized by rapid decomposition reducing their effectiveness. Colostrum of cows is a promising source of peptides with different functional orientations. One of the ways contributing to the expansion of using peptides when administered orally is their microcapsulation. The aim of the research is to develop a technology and method for microcapsulating peptides extracted from cow colostrum after calving and to assess their subchronic toxicity. The development of an installation for microencapsulation of peptides was carried out taking into account the diameter of the wide part of the cone of the working chamber and the speed of soaring of small particles of peptide conglomerates. For the experiment on the comparative assessment of the subchronic toxicity of encapsulated and free peptides, 4 groups of nonlinear mice were formed, 10 in each. Group 1 of mice was a control, group 2 (experimental) received an extract of the fabricium bag of broiler chickens containing peptides of various molecular weights at a dose of 15,000 mg/kg, group 3 – free peptides extracted from cow colostrum serum in an amount of 1,500 mg/kg, group 4 – encapsulated peptides in an amount of 15,000 mg/kg. Hematological blood analysis was performed using the analyzer PCE-90Vet. The mice were weighed on the 7th, 9th, 14th, 21st and 28th days of the experiment. Biochemical analysis of animal blood sera was carried out using the analyzer StatFax 3300. During the study, it has been found that the treatment of peptides with maltodextrin solution for 1.0–1.5 minutes forms a protective coating with a thickness of more than 2 microns. The diameters of the wide and narrow parts of the cone of the device and the velocity of the gushing streams for the smallest and largest particles of the peptide conglomerate have been calculated. The absence of toxicity of microcapsulated peptides has been proven.
(in Russian, стр.11, fig. 3, tables. 3, ref 25, AdobePDF, AdobePDF 0 Kb)
Vol. 28 (2025 year), No. 2, DOI: 10.21443/1560-9278-2025-28-2
Valieva Sh. S., Tikhonov S. L., Tikhonova N. V., Timofeeva M. S., Shikhalev S. V.
Construction of a proteolysis-resistant peptide sequence of the food recombinant protein GDF-11 and plasmid synthesis for its expression in E. coli
The GDF-11 protein has a positive effect on animals and humans in inflammatory and infectious processes, regeneration and rejuvenation, its high expression is observed in many organs and tissues. The relevance of the problem of oral use of many proteins and peptides, including the GDF-11 protein, as drugs and in specialized and functional products is noted. The aim of the research is to develop a proteolysis-resistant peptide sequence of the GDF-11 protein with virtual screening of its toxicity, allergenicity, uniqueness with subsequent synthesis of the corresponding plasmid for protein expression in E. coli. The GDF-11 protein, pET-25b(+) plasmids for expression in E. coli with the peptide sequence CTVDСFFECAFGСWDС have been used as objects of study. The GDF-11 protein gene with the sequence CTVDСFFECAFGСWDС has been synthesized from oligonucleotides using the cyclic assembly method. The obtained plasmid sequence encoding the new GDF-11 protein has been treated with restriction endonucleases BamHI and XhoI and cloned into the pET-25b(+) vector for E. coli expression. The clones have been sequenced by the Sanger method. The distribution and identification of primers in the obtained sequence have been studied by the MALDI-TOF MS Ultraflex method. A new unique peptide sequence CTVDСFFECAFGСWDС with cyclization at the C1-C4, C1-C2, C1-C3 bonds of the GDF-11 protein is developed, which allows increasing its resistance to proteolysis and, accordingly, opens up the possibility of oral use in prophylactic food products. Based on virtual screening, it is established that the new peptide chain CTVDСFFECAFGСWDС of the GDF-11 protein is not toxic to the cardiovascular and nervous systems, is not hepatotoxic, does not have cytotoxicity, does not have allergenicity and is characterized by high plasma clearance and an average half-life.
(in Russian, стр.8, fig. 4, tables. 2, ref 19, AdobePDF, AdobePDF 0 Kb)