Food Processing: Techniques and Technology Food Processing: Techniques and Technology Техника и технология пищевых производств 2074-9414 2313-1748 57772 10.21603/2074-9414-2023-1-2422 ОРИГИНАЛЬНАЯ СТАТЬЯ ORIGINAL ARTICLE ОРИГИНАЛЬНАЯ СТАТЬЯ Peptides of Trypsin Hydrolyzate in Bovine Colostrum Пептиды трипсинового гидролизата молозива коров https://orcid.org/0000-0003-4863-9834 Тихонов Сергей Леонидович Tikhonov Sergey L. tihonov75@bk.ru https://orcid.org/0000-0001-5841-1791 Тихонова Наталья Валерьевна Tikhonova Nataliya V. tihonov75@bk.ru Турсунов Хатам Хасанбаевич Tursunov Khatam Kh. https://orcid.org/0000-0001-6841-1197 Данилова Ирина Георгиевна Danilova Irina G. https://orcid.org/0000-0002-0470-7324 Лазарев Владимир Александрович Lazarev Vladimir A. Уральский государственный экономический университет Екатеринбург Россия Ural State University of Economics Yekaterinburg Russian Federation Уральский государственный лесотехнический университет Yekaterinburg RU Ural State Forestry Engineering University Yekaterinburg RU Уральский государственный экономический университет Екатеринбург Россия Ural State University of Economics Yekaterinburg Russian Federation Андижанский государственный медицинский институт Андижан Узбекистан Andijan State Medical Institute Andijan Uzbekistan Институт иммунологии и физиологии Уральского отделения Российской академии наук Екатеринбург Россия Institute of Immunology and Physiology of the Ural Branch of the Russian Academy of Sciences Yekaterinburg Russian Federation Уральский государственный экономический университет Екатеринбург Россия Ural State University of Economics Yekaterinburg Russian Federation 27 03 2023 27 03 2023 53 1 150 158 30 06 2022 04 10 2022 https://fptt.ru/en/issues/21374/21415/

Молозиво коров из-за содержания биологически активных веществ, в частности иммуноглобулинов, пептидов и цитокинов, является перспективным сырьем для производства продуктов функциональной направленности. Пептиды молозива обладают антимикробным действием. Биодоступность действующих начал молозива повышается при его ферментации протеолитическими ферментами. Цель исследования – выделение и характеристика пептидов надосадочной жидкости трипсиного гидролизата молозива коров, а также оценка их антимикробной и противогрибковой активностей. Для эксперимента использовали надосадочную жидкость трипсинового гидролизата молозива коров, выделенную методом центрифугирования при 3900 об/мин в течение 7 мин. Надосадочную жидкость разделяли методом препаративной хроматографии. Пептидный состав надосадочной жидкости ферментативного гидролизата определяли на МАЛДИ-ТОФ масс-спектрометре, расшифровку белковых последовательностей проводили с помощью базы данных Mascot. Для изучения белкового состава надосадочной жидкости гидролизата проводили осаждение белков сульфатом аммония. Антимикробную активность определяли диско-диффузионным методом. Культивирование штаммов бактерий проводили на плотной питательной среде LB при температуре 37 °C. Для оценки противомикробного действия пептидов провели эксперимент на крысах линии Вистар, инфицированных внутрибрюшинно Salmonella enteritidis 92. В надосадочном трипсиновом гидролизате молозива коров выделили 4 пептида, один из которых относится к коротким пептидам, три – к полипептидам. Выделенные пептиды имели различную молекулярную массу – 8,4, 6,5, 13,0 и 18 кДа. Установлено, что ферментативный гидролизат надосадочной жидкости молозива коров обладал бактерицидным действием к грамотрицательной бактерии Escherichia coli и грамположительной бактерии Bacillus subtilis, а также антигрибковой активностью против Candida albicans. Введение крысам, инфицированным S. enteritidis 92, внутрь трипсинового гидролизата надосадочной жидкости молозива коров способствовало их выживаемости, снижению ЛД50 и увеличению среднего срока гибели животных с 2 до 4 суток. Полученные данные свидетельствуют об антимикробном действии пептидов молозива и возможных иммуннотропных свойствах. Практическая значимость проведенного исследования заключается в перспективности использования пептидов надосадочной жидкости трипсиного гидролизата молозива коров для производства продуктов функциональной направленности с антимикробными свойствами.

Bovine colostrum contains biologically active substances, e.g., immunoglobulins, peptides, and cytokines, which makes it a logical component of numerous functional products. Colostrum peptides also possess antimicrobial activity. This bioavailability increases during colostrum fermentation with proteolytic enzymes. The research objective was to describe peptides isolated from the trypsic hydrolyzate supernatant of bovine colostrum and to evaluate their antimicrobial and antifungal properties. The supernatant of trypsin hydrolyzate of bovine colostrum was isolated by centrifugation at 3900 rpm for 7 min. The supernatant was separated by preparative chromatography. Its peptide composition was determined on a MALDI-TOF mass spectrometer, while the protein sequences were deciphered using the Mascot database. Proteins were precipitated with ammonium sulfate, and the antimicrobial activity was measured by the disk-diffusion method against gram-positive and gram-negative bacteria and dipoloid fungi. Strains were cultivated on a thick LB nutrient medium at 37°C. The antimicrobial activity was defined experimentally on Wistar rats infected intraperitoneally with Salmonella enteritidis 92. The trypsin hydrolyzate supernatant of bovine colostrum revealed four peptides, one of which belonged to short peptides, while the remaining three belonged to polypeptides. The isolated peptides had different molecular weights of 8.4, 6.5, 13.0, and 18 kDa. The enzymatic hydrolyzate proved bactericidal against Escherichia coli and Bacillus subtilis and demonstrated antifungal activity against Candida albicans. When rats infected with S. enteritidis 92 were administered with trypsin hydrolysate, it promoted their survival, decreased LD50, and increased the mean day of death period from 2 to 4 days. The research proved the antimicrobial effect of colostrum peptides and suggested their immunotropic properties. The peptides obtained from the trypsin hydrolyzate supernatant of bovine colostrum can be recommended for functional food industry as part of antimicrobial products.

 Молозиво молочный белок фермент гидролиз антимикробная активность противогрибковая активность биологически активные вещества Colostrum milk protein enzyme hydrolysis antimicrobial activity antifungal activity biologically active substances

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