Аннотация

Пептиды обладают высокой биологической активностью. Технологические режимы гидролиза белка определяют активность биопептидов. Целью исследования является выделение пептидов из фабрициевой сумки и оценка их иммуннотропного действия на мышах различных линий с экспериментальным иммунодефицитом путем изучения морфофункционального состояния органов иммунопоэза (тимус и селезенка).
В качестве объектов исследования выбраны гидролизат фабрициевой сумки, пептиды, тимус и селезенка мышей-самцов. Аминный азот определяли по ГОСТ Р 55479-2013, молекулярную массу пептидов – гельэлектрофорезом. Морфометрию тимуса и селезенки рассчитывали с помощью программы ВидеоТесТ-Морфология 5.0.
Определены рациональные режимы ферментативного гидролиза белка фабрициевой сумки папаином с последующим выделением пептидов методом ультрафильтрации. По содержанию аминного азота в гидролизате фабрициевой сумки определена рациональная концентрация фермента папаина – 0,15 %. Молекулярная масса пептидов после ультрафильтрации гидролизата фабрициевой сумки через мембраны с проницаемостью до 43 кДа распределяется в разных значениях и зависит от концентрации папаина, времени гидролиза и температуры. Максимальное количество пептидов с молекулярной массой от 27 до 18 кДа выделено при температуре гидролиза 36 °С (85 %), концентрации папаина 0,15 % (73 %), гидромодуле 1:3 (78 %) и времени гидролиза сырья 6 ч (82 %). Рациональные технологические параметры гидролиза белка фабрициевой сумки следующие: концентрация папаина 0,15 %, температура 36 °С, гидромодуль 1:3, время гидролиза 6 ч. Введение бурсальных пептидов иммунодефицитным мышам препятствует снижению общей клеточности в тимусе и уменьшению содержания CD3+ клеток, активизирует созревание Т-лимфоцитов и пролиферацию В-лимфоцитов, а также снижает иммуноцитотоксические эффекты от циклофосфамида.
Практическая значимость проведенного исследования заключается в перспективности использования разработанной технологии получения новых пептидов с выраженным иммунотропным действием и рассмотрении возможности их включения в рецептуру пищевой продукции специализированного назначения.

Ключевые слова

Пептиды, фабрициева сумка, гидролиз, тимус, селезенка, иммунодефицит, лабораторные животные

СПИСОК ЛИТЕРАТУРЫ

1. Kerksick СМ, Arent S, Schoenfeld BJ, Stout JR, Campbell B, Wilborn CD, et al. International society of sports nutrition position stand: nutrient timing. Journal of the International Society of Sports Nutrition. 2017;14(1). https://doi.org/10.1186/s12970-017-0189-4
2. Banihashemi SA, Nikoo M, Ghasempour Z, Ehsani A. Bioactive peptides fractions from traditional Iranian Koopeh cheese; lactic fermentation products. Biocatalysis and Agricultural Biotechnology. 2020;29. https://doi.org/10.1016/j.bcab.2020.101798
3. Meinlschmidt P, Sussmann D, Schweiggert-Weisz U, Eisner P. Enzymatic treatment of soy protein isolates: effects on the potential allergenicity, technofunctionality, and sensory properties. Food Science and Nutrition. 2016;4(1):11–23. https://doi.org/10.1002/fsn3.253
4. Pak VV, Koo MS, Kasymova TD, Kwon DY. Isolation and identification of peptides from soy 11S-globulin with hypocholesterolemic activity. Chemistry of Natural Compounds. 2005;41(6):710–714. https://doi.org/10.1007/s10600-006-0017-6
5. Marques MR, Fontanari GG, Pimenta DC, Soares-Freitas RM, Arêas JAG. Proteolytic hydrolysis of cowpea proteins is able to release peptides with hypocholesterolemic activity. Food Research International. 2015;77:43–48. https://doi.org/10.1016/j.foodres.2015.04.020
6. Ashraf J, Liu L, Awais M, Xiao T, Wang L, Zhou X, et al. Effect of thermosonication pre-treatment on mung bean (Vigna radiata) and white kidney bean (Phaseolus vulgaris) proteins: Enzymatic hydrolysis, cholesterol lowering activity and structural characterization. Ultrasonics Sonochemistry. 2020;66. https://doi.org/10.1016/j.ultsonch.2020.105121
7. Soares RAM, Mendonça S, de Castro LÍA, Menezes ACCCC, Arêas JAG. Major peptides from amaranth (Amaranthus cruentus) protein inhibit HMG-CoA reductase activity. International Journal of Molecular Sciences. 2015;16(2):4150–4160. https://doi.org/10.3390/ijms16024150
8. Prados IM, Plaza M, Marina ML, García M. Concepción evaluation of the relationship between the peptide profile and the lipid-lowering properties of olive seeds hydrolysates as a tool for tuning hypocholesterolemic functionality. Food and Function. 2020;11(6):4973–4981. https://doi.org/10.1039/d0fo00576b
9. Tsai J-S, Chen T-J, Pan BS, Gong S-D, Chung M-Y. Antihypertensive effect of bioactive peptides produced by protease-facilitated lactic acid fermentation of milk. Food Chemistry. 2008;106(2):552–558. https://doi.org/10.1016/j.foodchem.2007.06.039
10. Chen GW, Liao KJ. Method of combining pressure and enzyme to extract hydrolysate of animal-derived composition. Taiwan Patent No. TWI600379B. 2017.
11. Eisenmenger MJ, Reyes-De-Corcuera JI. High pressure enhancement of enzymes: A review. Enzyme and Microbial Technology. 2009;45(5):331–347. https://doi.org/10.1016/j.enzmictec.2009.08.001
12. Marciniak A, Suwal S, Naderi N, Pouliot Y, Doyen A. Enhancing enzymatic hydrolysis of food proteins and production of bioactive peptides using high hydrostatic pressure technology. Trends in Food Science and Technology. 2018;80:187–198. https://doi.org/10.1016/j.tifs.2018.08.013
13. Lin YH, Li CH, Lin CH, Yan YZ, Chuang YY, Chen GW. Application and development status of an ultra-high pressure processing technique in aquatic food. Fish. Ext. Rep. 2017;47:17–32.
14. Chen G-W, Yang M-H. Production and purification of novel hypocholesterolemic peptides from lactic fermented Spirulina platensis through high hydrostatic pressure-assisted protease hydrolysis. Catalysts. 2021;11(8). https://doi.org/10.3390/catal11080873
15. Anh TLQ, Hoa NTQ, Nguyen PDT, Van Thanh H, Nguyen PB, Anh LTH, et al. Soybean protein extraction by alcalase and flavourzyme, combining thermal pretreatment for enteral feeding product. Catalysts. 2020;10(8). https://doi.org/10.3390/catal10080829
16. Liu D-M, Chen J, Shi Y-P. Advances on methods and easy separated support materials for enzymes immobilization. TrAC – Trends in Analytical Chemistry. 2018;102:332–342. https://doi.org/10.1016/j.trac.2018.03.011
17. Kudryashov LS, Uzakov YaM, Tikhonov SL, Tikhonova NV, Diachkova AV. Microencapsulation of proteolytic enzymes for industrial application. News of the Academy of Sciences of the Republic of Kazakhstan. Series of Geology and Technical Sciences. 2020;3(441):161–169. https://doi.org/10.32014/2020.2518-170X.67
18. Tikhonov SL, Tikhonova NV, Kudryashov LS, Kudryashova OA, Moskovenko NV, Tretyakova IN. Efficiency of Microencapsulation of Proteolytic Enzymes. Catalysts. 2021;11(11). https://doi.org/10.3390/catal11111270
19. Zhang L, Lu Y, Feng X, Liu Q, Li Y, Hao J, et al. Hepatoprotective effects of Pleurotus ostreatus protein hydrolysates yielded by pepsin hydrolysis. Catalysts. 2020;10(6). https://doi.org/10.3390/catal10060595
20. Joshi I, Janagaraj K, Noorani KPM, Nazeer RA. Isolation and characterization of angiotensin I-converting enzyme (ACE-I) inhibition and antioxidant peptide from by-catch shrimp (Oratosquilla woodmasoni) waste. Biocatalysis and Agricultural Biotechnology. 2020;29. https://doi.org/10.1016/j.bcab.2020.101770
21. Narayanasamy A, Balde A, Raghavender P, Shashanth D, Abraham J, Joshi I, et al. Isolation of marine crab (Charybdis natator) leg muscle peptide and its anti-inflammatory effects on macrophage cells. Biocatalysis and Agricultural Biotechnology. 2020;25. https://doi.org/10.1016/j.bcab.2020.101577
22. Villadóniga C, Cantera AMB. New ACE-inhibitory peptides derived from α-lactalbumin produced by hydrolysis with Bromelia antiacantha peptidases. Biocatalysis and Agricultural Biotechnology. 2019;20. https://doi.org/10.1016/j.bcab.2019.101258
23. El-Sayed ST, Al-Azzouny RA, Ali OS. Purification and functional characterization of a novel tyrosinase (diphenolase) inhibitory peptides prepared from Solunum tuberosum peels protein via enzymatic hydrolysis. Biocatalysis and Agricultural Biotechnology. 2019;17:331–338. https://doi.org/10.1016/j.bcab.2018.12.009
24. Rani S, Pooja K, Pal GK. Exploration of potential angiotensin converting enzyme inhibitory peptides generated from enzymatic hydrolysis of goat milk proteins. Biocatalysis and Agricultural Biotechnology. 2017;11:83–88. https://doi.org/10.1016/j.bcab.2017.06.008
25. de Castro RJS, Sato HH. Simultaneous hydrolysis of proteins from different sources to enhance their antibacterial properties through the synergistic action of bioactive peptides. Biocatalysis and Agricultural Biotechnology. 2016;8:209–212. https://doi.org/10.1016/j.bcab.2016.09.014
26. Shafiei Kaleybar L, Khoshfetrat AB, Nozad Charoudeh H. Modeling and performance prediction of a conceptual bioprocess for mass production of suspended stem cells. Food and Bioproducts Processing. 2020;122:254–268. https://doi.org/10.1016/j.fbp.2020.04.012
27. Raja K, Martin LC, Bose L, Sahayanathan GJ, Padmanaban D, Chinnasamy A. Anti-proliferative and apoptotic effects of by-product (skin extract) from marine catfish Tachysurus dussumieri. Biocatalysis and Agricultural Biotechnology. 2020;29. https://doi.org/10.1016/j.bcab.2020.101816
28. Иммунотерапия: руководство для врачей. 2-е изд. / под ред. Р. М. Хаитова, Р. И. Атауллаханова, А. Е. Шульженко. М.: ГЭОТАР-Медиа, 2018. 767 с.
29. Xu Q, Hong H, Wu J, Yan X. Bioavailability of bioactive peptides derived from food proteins across the intestinal epithelial membrane: A review. Trends in Food Science and Technology. 2019;86:399–411. https://doi.org/10.1016/j.tifs.2019.02.050
30. Травникова Н. А. Сравнительная морфология фабрициевой бурсы цыплят-бройлеров в возрастном аспекте при разных способах содержания: автореф. дис…канд. вет. наук. Екатеринбург, 2004. 22 с.
31. Li D, Xue M, Geng Z, Chen P. The suppressive effects of bursopentine (BP5) on oxidative stress and NF-ĸB activation in lipopolysaccharide-activated murine peritoneal macrophages. Cellular Physiology and Biochemistry. 2012;29:9–20. https://doi.org/10.1159/000337581
32. Li J, Li T-X, Ma Y, Zhang Y, Li D-Y, Xu H-R. Bursopentin (BP5) induces G1 phase cell cycle arrest and endoplasmic reticulum stress/mitochondria-mediated caspase-dependent apoptosis in human colon cancer HCT116 cells. Cancer Cell International. 2019;19(1). https://doi.org/10.1186/s12935-019-0849-3
33. Ahangari H, Yazdani P, Ebrahimi V, Soofiyani SR, Azargun R, Tarhriz V, et al. An Updated review on production of food derived bioactive peptides; focus on the psychrotrophic bacterial proteases. Biocatalysis and Agricultural Biotechnology. 2021;35. https://doi.org/10.1016/j.bcab.2021.102051
34. Dullius A, Goettert MI, de Souza CFV. Whey protein hydrolysates as a source of bioactive peptides for functional foods – Biotechnological facilitation of industrial scale-up. Journal of Functional Foods. 2018;42:58–74. https://doi.org/10.1016/j.jff.2017.12.063
35. Martínez-Sánchez SM, Gabaldón-Hernández JA, Montoro-García S. Unravelling the molecular mechanisms associated with the role of food-derived bioactive peptides in promoting cardiovascular health. Journal of Functional Foods. 2020;64. https://doi.org/10.1016/j.jff.2019.103645
36. Khaitov RM. Immunomodulators: myths and reality. Immunologiya. 2020;41(2):101–106. https://doi.org/10.33029/0206-4952-2020-41-2-101-106
37. Liu X-D, Feng X-L, Zhou B, Cao R-B, Li X-F, Ma Z-Y, et al. Isolation, modulatory functions on murine B cell development and antigen-specific immune responses of BP11, a novel peptide from the chicken bursa of Fabricius. Peptides. 2012;35(1):107–113. https://doi.org/10.1016/j.peptides.2012.03.003
38. Yin Y, Qin T, Yu Q, Yang Q. Bursopentin (BP5) from chicken bursa of fabricius attenuates the immune function of dendritic cells. Amino Acids. 2014;46(7):1763–1774. https://doi.org/10.1007/s00726-014-1735-x
39. Feng X-L, Zhou B, Cao R-B, Liu Q-T, Liu K, Liu X-D, et al. Immunomodulatory roles and functional analysis of pre-B lymphocyte DT40 cells with the bursal-derived BSP-II treatment. Peptides. 2012;36(2):292–298. https://doi.org/10.1016/j.peptides.2012.04.015