chemicallytechFine Chemical TechnologiesТонкие химические технологии2410-65932686-7575MIREA – Russian Technological University (RTU MIREA).10.32362/2410-6593-2026-21-2-179-187NEFNGTchemicallytech-2386Research ArticleCHEMISTRY AND TECHNOLOGY OF MEDICINAL COMPOUNDS AND BIOLOGICALLY ACTIVE SUBSTANCESХИМИЯ И ТЕХНОЛОГИЯ ЛЕКАРСТВЕННЫХ ПРЕПАРАТОВ И БИОЛОГИЧЕСКИ АКТИВНЫХ СОЕДИНЕНИЙEnzymatic deglycosylation of soy proteins as a method to increase the efficiency of their hydrolysisФерментативное дегликозилирование соевых белков как способ повышения эффективности их гидролизаhttps://orcid.org/0000-0001-5348-4350ЛеонтьевВ. Н.LeontievV. N.

Леонтьев Виктор Николаевич, к.х.н., заведующий кафедрой биотехнологии

Scopus Author ID 14052100400

220006, Минск, ул. Свердлова, д. 13а

Viktor N. Leontiev, Cand. Sci. (Chem.), Head of the Department of Biotechnology

Scopus Author ID 14052100

13a, Sverdlova ul., Minsk, 220006

leontiev@belstu.byhttps://orcid.org/0009-0006-0919-3736ЛазовскаяО. И.LazovskayaO. I.

Лазовская Олеся Илгамовна, ассистент, кафедра биотехнологии

Scopus Author ID 57205667523

220006, Минск, ул. Свердлова, д. 13а

Olesya I. Lazovskaya, Assistant, Department of Biotechnology

Scopus Author ID 57205667523

13a, Sverdlova ul., Minsk, 220006

lazovskaya@belstu.byБелорусский государственный технологический университетБеларусьBelarusian State Technological UniversityBelarus202606052026212179187Copyright © Leontiev V.N., Lazovskaya O.I., 20262026Леонтьев В.Н., Лазовская О.И.Leontiev V.N., Lazovskaya O.I.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.finechem-mirea.ru/jour/article/view/2386Objectives

Objectives. Soy protein hydrolysates are now widely used in the food industry, fish farming, poultry farming, livestock farming, as well as in medical preparations. The most effective method for their production is enzymatic hydrolysis. However, even with optimal proteolysis parameters, it is not always possible to achieve the required degree of hydrolysis. For this reason, various technological approaches are used to more intensively break down soy proteins, including the addition of enzyme preparations and pretreatment of the protein substrate. β-Conglycinin, one of the main soy proteins, is a glycoprotein whose carbohydrate portion consists primarily of mannose residues. We hypothesize that deglycosylation of β-conglycinin by an enzyme preparation with mannanase activity as a pretreatment of the soy substrate will lead to change in the structure of its protein portion due to the destruction of the carbohydrate component to increase the accessibility of peptide bonds to proteolytic enzymes. Thus, the work sets out to study the effect of enzymatic deglycosylation on the efficiency of soy protein hydrolysis.

Methods

Methods. Deglycosylation of β-conglycinin, hydrolysis of polysaccharides and lipids were performed by the Complex-concentrate enzyme preparation (Ferment, Republic of Belarus). Protein hydrolysis was carried out by the Protozyme C330 enzyme preparation (Ferment, Republic of Belarus). The formation of reducing sugars was confirmed by the Miller method. The degree of protein hydrolysis was determined by the pH-stat method. The molecular weight distribution of peptide fractions was analyzed by low-pressure liquid gel chromatography on a column with Sephadex® G-50 Medium. Computer processing of the elution profile of peptide fractions was performed in the OriginPro 8.5.1 program using the Gauss function.

Results

Results. It is established that the treatment of soy flour by the Complex-concentrate enzyme preparation (enzyme-substrate ratio 1 : 40, hydromodule 1 : 10) promotes the breakdown of both free oligo- and polysaccharides, as well as the carbohydrate component β-conglycinin. Proteolysis by the Protozyme C330 enzyme preparation (enzyme-substrate ratio 1 : 20, pH 7.5, 50°C, 3.5 h) carried out following 20 h of deglycosylation results in a product with a degree of hydrolysis of 56.3%. The content of low-molecular-weight peptides in soy hydrolysate is 83.9%. Proteolysis without enzymatic destruction of the carbohydrate part of β-conglycinin is shown to be characterized by a degree of hydrolysis of 9.2%.

Conclusions

Conclusions. A pretreatment approach involving deglycosylation of enzymatic β-conglycinin can be used to significantly increase the degree of hydrolysis of soy proteins.

Цели

Цели. В настоящее время гидролизаты соевых белков находят широкое применение в пищевой промышленности, медицине, рыбоводстве, птицеводстве и животноводстве. Наиболее эффективным способом их получения является ферментативный гидролиз. Однако даже при оптимальных параметрах протеолиза не всегда возможно достичь требуемой степени гидролиза, поэтому для более интенсивного расщепления соевых белков используют различные технологические подходы: внесение нескольких ферментных препаратов и предварительную обработку белкового субстрата. β-Конглицинин — один из основных белков сои — представляет собой гликопротеин, углеводная часть которого состоит преимущественно из маннозных остатков. Предполагаем, что дегликозилирование β-конглицинина ферментным препаратом с маннаназной активностью в качестве предварительной обработки соевого субстрата приведет к изменению структуры его белковой части за счет разрушения углеводного компонента и позволит повысить доступность пептидных связей к действию протеолитических ферментов. Таким образом, целью работы является изучение влияния ферментативного дегликозилирования на эффективность гидролиза соевых белков.

Методы

Методы. Дегликозилирование β-конглицинина, гидролиз полисахаридов и липидов проводили ферментным препаратом «Комплекс-концентрат» (ООО «Фермент», Республика Беларусь). Гидролиз белков осуществляли ферментным препаратом «Протозим С330» (ООО «Фермент», Республика Беларусь). Образование редуцирующих сахаров подтверждали методом Миллера. Степень гидролиза белков определяли pH-статическим методом. Молекулярно-массовое распределение пептидных фракций анализировали методом жидкостной гель-хроматографии низкого давления на колонке с гелем Sephadex® G-50 Medium. Компьютерную обработку профиля элюирования пептидных фракций выполняли в программе OriginPro 8.5.1 с помощью функции Гаусса.

Результаты

Результаты. Установлено, что обработка соевой муки ферментным препаратом «Комплекс-концентрат» (фермент-субстратное соотношение 1 : 40, гидромодуль 1 : 10) способствует расщеплению как свободных олиго- и полисахаридов, так и углеводного компонента β-конглицинина. Протеолиз ферментным препаратом «Протозим С330» (фермент-субстратное соотношение 1 : 20, рН 7.5, 50°C, 3.5 ч) после 20-часового дегликозилирования приводит к получению продукта со степенью гидролиза 56.3%. При этом содержание низкомолекулярных пептидов в соевом гидролизате составляет 83.9%. Показано, что протеолиз без ферментативного разрушения углеводной части β-конглицинина характеризуется степенью гидролиза 9.2%.

Выводы

Выводы. Применение ферментативного дегликозилирования β-конглицинина в качестве предварительной обработки позволяет существенно повысить степень гидролиза соевых белков.

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The authors declare that there are no conflicts of interest present.