spfpХранение и переработка сельхозсырьяStorage and Processing of Farm Products2072-96692658-767XРОСБИОТЕХ10.36107/spfp.2022.365spfp-365Research ArticleБИОТЕХНОЛОГИЧЕСКИЕ И МИКРОБИОЛОГИЧЕСКИЕ АСПЕКТЫBIOTECHNOLOGICAL AND MICROBIOLOGICAL ASPECTSО введении принципа насыщающей дополнительности ферментативного процесса в методологию глубокой переработки растительного сырьяIntroduction the Principle of Saturation Additionality for Enzymatic Process into the Methodology of Plants Raw Materials Complete Processinghttps://orcid.org/0000-0001-9879-482XПетровАндрей НиколаевичPetrovAndrey N.petrovmilk@mail.ruhttps://orcid.org/0000-0001-8237-0774КондратенкоТатьяна ЮрьевнаKondratenkoTatyana Yu.t.kondratenko@fncps.ruВсероссийский научно-исследовательский институт технологии консервирования – филиал ФГБНУ «Всероссийский научный центр пищевых систем им. В.М. Горбатова» РАН, кандидат технических наук, 142703, Россия, Московская область, Ленинский городской округ, г. Видное, ул. Школьная, д. 78РоссияRussian Research Institute of Canning Technology – branch of Gorbatov Research Center for Food SystemsRussian Federation2022031020220393108Copyright © Петров А.Н., Кондратенко Т.Ю., 20222022Петров А.Н., Кондратенко Т.Ю.Petrov A.N., Kondratenko T.Y.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.spfp-mgupp.ru/jour/article/view/365

Введение: Биотехнологический подход к глубокой переработке растительного сырья с использованием ферментных препаратов позволяет эффективно использовать нативный биологический и/или технологический потенциал. Целевые компоненты сырья являются фрагментами молекулярных компонентов матрикса клеточных стенок с трудно устанавливаемой концентрацией целевых гликозидных связей, которая необходима для определения кинетических характеристик ферментных препаратов. Материалы и методы: Объектом исследования был негранулированный сухой немелассированный свекловичный жом, а также отечественные ферменты лиазного и гидролазного действия. В работе использован подход, основанный на аппроксимации экспериментальных данных с последующим определением горизонтальных асимптот. Цель: Обосновать адекватность применения косвенных показателей, таких как удельная электрическая проводимость, при оценке кинетических показателей ферментных препаратов лиазного и гидролазного действия. Результаты: Получен массив экспериментальных данных динамик удельной электрической проводимости от времени при обработке свекловичного жома ферментными препаратами в интервале концентраций от 0 до 0,8 %. В результате аппроксимации рассчитаны локальные пределы концентрации субстрата, выраженные в косвенных единицах. Предложен комплекс постулатов динамики системы «субстрат – ферментный препарат», на основании которого сформирован принцип насыщающей дополнительности ферментативного процесса, согласно которому локальный предел концентрации субстрата, достигаемый при данной концентрации фермента, составляет дробную часть некоторого глобального предела концентрации, который может быть полностью переведён в продукт посредством нескольких этапов, локальный предел продолжительности каждого из которых стремится к бесконечности. Экспериментально установлено, что в случае применения ферментных препаратов лиазного и гидролазного действия, рассчитанные локальные пределы концентрации субстрата монотонно увеличиваются при увеличении концентрации ферментного препарата, вырождаясь в горизонтальную асимптоту, соответствующую глобальному пределу концентрации субстрата, что подтверждает как сам принцип насыщающей дополнительности, так и следствия из него. Экспериментально установлена применимость малых концентраций ферментных препаратов (в пределах 0,1-0,2 %) и нескольких последовательных этапов для ферментативной трансформации субстрата в продукт. Выводы. В результате проведённых исследований был разработан принцип насыщающей дополнительности ферментативного процесса, позволяющий на основе экспериментальных данных по динамикам ферментативной трансформации обрабатываемого объекта ферментными препаратами в заданных концентрациях однозначно определить эффективную концентрацию целевого субстрата в условиях отсутствия определённости относительно её численного значения, либо невозможности прямого определения. Результирующее значение может быть использовано для установления кинетических характеристик ферментативного процесса, таких как Vmax и Km. Принцип насыщающей дополнительности применим в отношении гомоферментных препаратов лиазного и гидролазного действия и является составляющей дерева принятия решений для разработки технологий промышленного производства растительных полигликанов.

Background: Biotechnological approach to complete processing of plant raw materials using enzyme preparations allows the efficient use of its native biological and/or technological potential. The point components are fragments of molecular components of the cell wall matrix with a target glycoside bonds' hard-to-find concentration which is necessary to determine the enzyme kinetic characteristics. Materials and methods: Non-granulated dry non-molassed sugar beet pulp as well as lyase and hydrolase enzymes were used as research objects Purpose: to study an approach based on approximation of experimental data followed by determination of horizontal asymptotes. Results: The adequacy of using an indirect indicators, such as specific electrical conductivity, for assessing the kinetic parameters of lyase and hydrolase enzymes  is substantiated. An array of experimental data was obtained for dynamics of specific electrical conductivity during processing beet pulp with enzyme preparations in the concentration range from 0 to 0.8%. As a result of the approximation, the local limits of the substrate concentration were calculated in indirect units. For dynamic of the "substrate - enzyme" system, a set of postulates was proposed. On the basis of this set, the principle of saturating complementarity for enzymatic process is formed. In according with it the local substrate concentration limit achieved at a given enzyme concentration is a fractional part of some global one, which can be completely converted into a product through several stages where the local limit of the duration of each stage tends to infinity. It has been experimentally established that, in the case of lyase and hydrolase enzyme, the calculated local substrate concentration limits increase monotonously with an increase of enzyme concentration, degenerating into a horizontal asymptote corresponding to the global substrate concentration limit, which confirms both the principle of saturating complementarity and its consequences. The applicability of low enzyme concentrations (within 0.1-0.2%) and several step-by-step stages for the enzymatic transformation of a substrate into a product has been experimentally established. Conclusion: As a result of research, the principle of saturating complementarity for enzymatic process was developed. Based on experimental data on the enzymatic transformation dynamics for processed object, it allows an unambiguous determination of the effective substrate concentration in the absence of certainty about its quantity value, or the impossibility of direct determination. The resulting value can be used to establish the kinetic characteristics of enzymatic process, such as Vmax and Km. The principle of saturating complementarity is applicable for lyase and hydrolase homoenzymes. It is a one of the necessary components within the decision tree for development the technologies for industrial production of plant polyglycans.

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