Food Processing: Techniques and Technology Food Processing: Techniques and Technology Техника и технология пищевых производств 2074-9414 2313-1748 92201 10.21603/2074-9414-2024-4-2533 ОРИГИНАЛЬНАЯ СТАТЬЯ ORIGINAL ARTICLE ОРИГИНАЛЬНАЯ СТАТЬЯ Lactulose Biosynthesis with Enzymes of Lactose-Fermenting Micromycetes and Bacteria Биосинтез лактулозы с использованием ферментов лактозосбраживающих микромицет и бактерий https://orcid.org/0000-0001-9803-8709 Рябцева Светлана Андреевна Ryabtseva Svetlana A. ryabtseva07@mail.ru https://orcid.org/0000-0002-0119-9061 Шпак Мария Александровна Shpack Maria A. https://orcid.org/0000-0001-8460-2954 Лодыгин Алексей Дмитриевич Lodygin Alexei D. allodygin@yandex.ru Серов Александр Владимирович Serov Alexander V.

доктор технических наук;

doctor of technical sciences;

 https://orcid.org/0000-0002-8200-3007 Сазанова Серафима Николаевна Sazanova Serafima N. https://orcid.org/0009-0009-9593-6074 Скороходова Марина Владимировна Skorokhodova Marina V. https://orcid.org/0009-0000-1066-4394 Ромахова Вера Юрьевна Romakhova Vera Yu.

кандидат биологических наук;

candidate of sciences in biology;

 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 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 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 Северо-Кавказский федеральный университет Ставрополь Россия North-Caucasus Federal University Stavropol Russian Federation 24 12 2024 24 12 2024 54 4 645 657 14 03 2024 07 05 2024 https://fptt.ru/en/issues/23109/23111/

Лактулоза относится к хорошо изученным пребиотикам и давно используется в медицине и пищевой промышленности. Альтернативой традиционным химическим способам является получение лактулозы с применением бета-галактозидаз.Целью работы было исследование процессов биосинтеза лактулозы с использованием неочищенных ферментных препаратов β-галактозидаз, полученных при раздельном и совместном культивировании лактозосбраживающих дрожжей и молочнокислых бактерий в пермеате молочной сыворотки. В качестве продуцентов бета-галактозидаз использовали штаммы лактозосбраживающих дрожжей Kluyveromyces lactis ВКМ Y-1333 и Y-1339, Kluyveromyces marxianus ВКМ Y-459 и Y-1338, а также вязкие расы молочнокислых бактерий Lactobacillus acidophilus БК-Углич-АВ и Streptococcus thermophilиs БК-Углич-ТВ. Определение активности β-галактозидазы было основано на гидролизе о-нитрофенил-β-D-галактопиранозида. Определение углеводного состава проводили методом высокоэффективной жидкостной хроматографией на хроматографе SHIMADZU LC-20 Prominence. К общим закономерностям образования лактулозы при использовании ферментов исследованных дрожжей можно отнести быстрый рост ее концентрации в течение первых 30 мин реакции с последующим снижением или стабилизацией. Активность некоторых комбинированных неочищенных ферментных препаратов дрожжей и молочнокислых микроорганизмов существенно превышает активность бета-галактозидаз продуцентов после их раздельного культивирования. Закономерности образования лактулозы зависят как от штамма дрожжей, так и от вида молочнокислых бактерий. Самые высокие выходы лактулозы были получены при использовании комбинированных β-галактозидаз всех исследованных штаммов дрожжей с вязкими расами S. thermophilus. Совместное культивирование дрожжей и молочнокислых бактерий в некоторых сочетаниях позволило получить комбинированные неочищенные ферментные препараты с более высокой активностью и более высокие выходы лактулозы, чем отдельное культивирование дрожжей. Данные результаты могут быть использованы для получения очищенных ферментных препаратов β-галактозидаз и лактулозосодержащих продуктов из вторичного мол очного сырья.

Lactulose is a well-studied prebiotic popular in medicine and the food industry. β-Galactosidases offer an alternative to traditional chemical methods of lactulose production. The article describes lactulose biosynthesis with crude enzyme preparations of β-galactosidases obtained by separate and joint cultivation of lactose-fermenting yeasts and lactic acid bacteria in whey permeate. The research featured the following producers of β-galactosidases: lactose-fermenting yeasts of Kluyveromyces lactis VKM Y-1333 and Y-1339, Kluyveromyces marxianus VKM Y-459 and Y-1338, and viscous strains of lactic acid bacteria of Lactobacillus acidophilus and Streptococcus thermophilis. The β-galactosidase activity was measured using the method of o-nitrophenyl-β-D-galactopyranoside hydrolysis. The method of high-performance liquid chromatography made it possible to reveal the carbohydrate composition. The lactulose formation by yeast enzymes demonstrated the following pattern: its concentration increased rapidly during the first 30 min of reaction with subsequent decrease or stabilization. Some combined unpurified enzyme preparations of yeasts and lactic acid microorganisms were more active than β-galactosidases obtained by separate cultivation. The patterns of lactulose formation depended on both the yeast strain and the type of lactic acid bacteria. The highest yields of lactulose belonged to the samples that combined β-galactosidases of all yeast strains with Streptococcus thermophilis. Co-cultivation of yeast and lactic acid bacteria in some combinations produced combined crude enzyme preparations with higher activity and greater lactulose yields than separate yeast cultivation. These results can help to obtain purified enzyme preparations of β-galactosidases and lactulose-containing products from secondary dairy raw materials.

 Лактулоза лактоза трансгликозилирование β-галактозидазы Kluyveromyces marxianus Kluyveromyces lactis Lactobacillus acidophilus Streptococcus thermophilus Lactulose lactose transglycosylation β-galactosidases Kluyveromyces marxianus Kluyveromyces lactis Lactobacillus acidophilus Streptococcus thermophilus Работа выполнена при финансовой поддержке Министерства науки и высшего образования Российской Федерации в рамках реализации комплексного проекта по созданию высокотехнологичного производства по теме: «Создание первого в России высокотехнологичного производства пребиотика лактулозы и функциональных молочных ингредиентов для импортозамещения в медицине, ветеринарии, детском питании, производстве лечебно-профилактических продуктов для людей и животных» (Соглашение о предоставлении из федерального бюджета субсидии на развитие кооперации государственного научного учреждения и организации реального сектора экономики в целях реализации комплексного проекта по созданию высокотехнологичного производства №075-11-2022-021 от 07.04.2022 г.) в рамках Постановления Правительства РФ от 9 апреля 2010 г. № 218 на базе ФГАОУ ВО «Северо-Кавказский федеральный университет ». The research was supported by t he Ministry of Science and Higher Education of the Russian Federation as part of a comprehensive high-tech project of First Russian high-tech lactulose prebiotic and functional dairy ingredients for import substitution in medicine, veterinary practice, baby food, and medical preventive products for humans and animals (Agreement No. 075-11-2022-021, April 07, 2022, on federal budget subsidies for cooperation between state scientific institutions and the real sector of the economy in the sphere of high-tech production); Decree No. 218, April 9, 2010, of the Government of the Russian Federation to the North Caucasian Federal University .

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