Food Processing: Techniques and Technology Food Processing: Techniques and Technology Техника и технология пищевых производств 2074-9414 2313-1748 88729 10.21603/2074-9414-2024-3-2521 ОРИГИНАЛЬНАЯ СТАТЬЯ ORIGINAL ARTICLE ОРИГИНАЛЬНАЯ СТАТЬЯ Biologically Active Dairy Protein Hydrolysates and Their Cyclodextrin Inclusion Complexes: Biotechnological Aspects of Production Биологически активные гидролизаты белков молока и их комплексы включения с циклодектринами https://orcid.org/0000-0002-2096-8030 Головач Татьяна Николаевна Halavach Tatsiana M. halavachtn@gmail.com https://orcid.org/0000-0002-2641-3888 Ловкис Зенон Валентинович Lovkis Zenon V. https://orcid.org/0000-0002-4859-2389 Курченко Владимир Петрович Kurchenko Vladimir P. kurchenko@tut.by https://orcid.org/0000-0001-6786-3589 Янцевич Алексей Викторович Yantsevich Aleksei V. https://orcid.org/0000-0001-5711-6037 Тарун Екатерина Ивановна Tarun Ekaterina I. https://orcid.org/0000-0002-5877-9307 Дудчик Наталья Владимировна Dudchik Natalia V. https://orcid.org/0000-0002-2632-8923 Ржепаковский Игорь Владимирович Rzhepakovsky Igor V. https://orcid.org/0000-0001-8460-2954 Лодыгин Алексей Дмитриевич Lodygin Alexei D. allodygin@yandex.ru Белорусский государственный университет Минск Беларусь Belarusian State University Minsk Belarus Научно-практический центр Национальной академии наук Беларуси по продовольствию Минск Беларусь Scientific and Practical Centre for Foodstuffs of the National Academy of Sciences of Belarus Minsk Belarus Белорусский государственный университет Минск Беларусь Belarusian State University Minsk Belarus Институт биоорганической химии Национальной академии наук Беларуси Минск Беларусь Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus Minsk Belarus Международный государственный экологический институт имени А.Д. Сахарова Минск Беларусь International Sakharov Environmental Institute of Belarusian State University Minsk Belarus Республиканское унитарное предприятие «Научно-практический центр гигиены» Минск Беларусь Scientific and Practical Centre of Hygiene Minsk Belarus Северо-Кавказский федеральный университет Ставрополь Россия North-Caucasus Federal University Stavropol Russian Federation Северо-Кавказский федеральный университет Ставрополь Россия North-Caucasus Federal University Stavropol Russian Federation 02 10 2024 02 10 2024 54 3 461 482 19 03 2024 07 05 2024 https://fptt.ru/en/issues/22856/22817/

Ферментативный гидролиз белков молока – эффективный технологический прием для повышения их пищевой и биологической ценности, а также снижения аллергенного потенциала. Последующее комплексообразование пептидов с циклодекстринами (ЦД) направлено на снижение горечи гидролизованных белков. Целью работы являлось получение гидролизатов сывороточных белков молока и комплексов включения пептидов с ЦД, определение пептидного состава расщепленных белков молока, оценка биоактивных и органолептических свойств гидролизатов и комплексов включения. Объекты исследования: ферментативные гидролизаты сывороточных белков молока с глубокой степенью гидролиза и их комплексы включения с β- и γ-ЦД. Для расщепления белков применяли протеазу алкалазу (Alcalase®2.4L FG), гидролизаты подвергали микро- и ультрафильтрации (отсечка 10 кДа). Пептидный состав гидролизованных белков определяли высокоэффективной жидкостной хроматографией и хромато-масс-спектрометрией, антимутагенную активность – в тесте Эймса, антибактериальное действие – импедиметрическим методом. Антиоксидантный эффект оценивали с использованием флуориметрического и спектрофотометрического подходов, антигенные свойства – по данным конкурентного иммуноферментного анализа. Уровень горечи опытных образцов устанавливали органолептически. Подобраны режимы расщепления сывороточных белков молока алкалазой для достижения эффективной микро- и ультрафильтрации гидролизата. Получены гипоаллергенные пептидные фракции и их комплексы включения с β- и γ-ЦД. В результате протеолиза молочной сыворотки и последующей фильтрации с/без тиндализации установлено снижение остаточной антигенности в 265/589 раз соответственно. По данным флуориметрического метода показано увеличение антиоксидантной активности гидролизата в составе комплексов с β/γ-ЦД в 1,79/1,90 раза соответственно. Связывание β-ЦД с пептидами обусловило возрастание их антимикробного действия в отношении Escherichia coli АТСС 8739 и Staphylococcus aureus АТСС 6538. Эффективное снижение горечи установлено после взаимодействия гидролизата с β-ЦД. Ферментативные гидролизаты белков молока и их комплексы включения с ЦД являются перспективными заменителями нативного белкового компонента для продуктов питания функционального назначения, что обусловлено их низким аллергенным потенциалом, приемлемым вкусом и подтвержденными биоактивными свойствами.

Enzymatic hydrolysis of dairy proteins increases their nutritional and biological value while reducing their allergenic potential. The subsequent complexation of peptides with cyclodextrins (CDs) reduces the bitterness of the hydrolyzed proteins. The research objective was to obtain hydrolysates of whey proteins and their cyclodextrin inclusion complexes with peptides, as well as to describe the peptide composition of the cleaved dairy proteins, biological activity, and sensory profile of the hydrolysates and inclusion complexes. The research featured enzymatic whey protein hydrolysates with an extensive hydrolysis degree and their inclusion complexes with β- and γ-CDs. Dairy proteins were hydrolyzed with alcalase, and the hydrolysates obtained were subjected to micro- and ultrafiltration (cut-off limit – 10 kDa). The peptide composition of the hydrolyzed proteins was determined by the methods of high-performance liquid chromatography and chromatography-mass spectrometry. The antimutagenic activity was evaluated using the Ames test whereas the antibacterial effect was studied with the impedimetric method. The antioxidant activity was detected with fluorimetry and spectrophotometry. The method of competitive enzyme immunoassay was applied to reveal the antigenic properties. The bitterness of the experimental sample s was determined by a sensory evaluation. The research delivered the optimal modes for whey protein cleavage with alcalase that made it possible to achieve efficient micro- and ultrafiltration. The resulting hypoallergenic peptide fractions and their inclusion complexes with β- and γ-CDs possessed antioxidant, antibacterial, and antimutagenic properties. The whey proteolysis and subsequent filtration with/without tindalization demonstrated a 265/589-fold decrease in the residual antigenicity. The fluorimetric method showed a 1.79/1.90-fold increase in the antioxidant activity of the hydrolysate in complexes with β- and γ-CDs. Binding of β-CDs to peptides enhanced their antimicrobial activity against Escherichia coli ATCC 8739 and Staphylococcus aureus ATCC 6538. The hydrolysate samples with β-CDs showed less bitterness. Whey proteolysis with alcalase under optimized conditions and subsequent fractionation resulted in a product with high consumer qualities. Enzymatic hydrolysates of dairy proteins and their CD inclusion complexes were able to substitute native protein components. Their bioactive properties, good taste, and low allergenic potential mean good prospects for the functional food industry.

 Сывороточные белки ферментативный гидролиз ультрафильтрация степень гидролиза пептиды циклодекстрин биологическая активность Whey proteins enzymatic hydrolysis ultrafiltration hydrolysis degree peptide cyclodextrin biological activity Работа выполнена в рамках гранта Министерства образования Республики Беларусь «Нанокомплексы циклодекстринов с биологически активными гидролизатами белков молока для специализированных продуктов питания» (договор № 002/2023–БГУ от 20.03.2023) и «Биологически активные фракции гидролизатов белков молока для продуктов функционального назначения» (договор № 002/2024–БГУ от 13.03.20 24) The research was supported by the Ministry of Education of Belarus as part of grant no. 002/2023-BSU: Nanocomplexes of cyclodextrins with biologically active hydrolysates of dairy proteins for specialty foods (20.03.2023) and no. 002/2024-BSU: Biologically active fractions of dairy protein hydrolysate s for functional products (13.03.2024)

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