spfpХранение и переработка сельхозсырьяStorage and Processing of Farm Products2072-96692658-767XРОСБИОТЕХ10.36107/spfp.2023.426spfp-426Research ArticleБИОТЕХНОЛОГИЧЕСКИЕ И МИКРОБИОЛОГИЧЕСКИЕ АСПЕКТЫBIOTECHNOLOGICAL AND MICROBIOLOGICAL ASPECTSОпределение целесообразности предварительной декатионизации растительного сырья для ферментативной фрагментации протопектинового комплексаDetermination of the Feasibility of Preliminary Decationization of Plant Raw Materials for Enzymatic Fragmentation of the Protopectin Complexhttps://orcid.org/0000-0002-0913-5644КондратенкоВладимир ВладимировичKondratenkoVladimir V.

SPIN-код: 3383-1774;  Researcher ID: E-3592-2010

 

 

v_kondratenko@vnimi.orghttps://orcid.org/0000-0001-8237-0774КондратенкоТатьяна ЮрьевнаKondratenkoTatyana Yu.

SPIN-код: 7558-8832;  Researcher ID: ABF-3810-2022

t.kondratenko@fncps.ruВсероссийский научно-исследовательский институт молочной промышленности»РоссияRussian Dairy Research InstituteRussian FederationВсероссийский научный центр пищевых систем им. В.М. ГорбатоваРоссияRussian Research Institute of Canning Technology — branch of Gorbatov Research Center for Food Systems of the Russian Academy of SciencesRussian Federation20233006202302118131Copyright © Кондратенко В.В., Кондратенко Т.Ю., 20232023Кондратенко В.В., Кондратенко Т.Ю.Kondratenko V.V., 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/426Введение

Введение: Использование ферментных препаратов для обработки растительного сырья налагает некоторые ограничения, определяемые структурой молекул ферментов и свойствами среды. Одним из основных лимитирующих факторов является возможное присутствие в среде ингибитора, подавляющего каталитическую активность. Потенциальным ингибитором могут являться катионы поливалентных металлов, поэтому определение целесообразности декатионизации среды перед ферментативной обработкой является актуальным.

Цель

Цель: исследования является разработка подхода к определению целесообразности предварительной декатионизации сырья перед ферментативным процессом.

Материалы и методы

Материалы и методы: В качестве субстрата использовали негранулированный сухой немелассированный свекловичный жом. Ферментативную обработку проводили гомоферментным препаратом рамногалактуронан лиазного действия в концентрации 0,00,0,05, 0,1, 0,2, 0,4 и 0,8 % от массы сухого субстрата. Предварительную обработку набухшего сырья поводили 0,00, 0,20, 0,50 и 0,91 ед. от 2,87 % водного раствора комплексона (ЭДТА). Фактором отклика была удельная электрическая проводимость. Ферментативный процесс проводили при температуре 45 °С течение 8 часов. Аппроксимацией экспериментальных данных для каждого варианта исследований получали математическое описание динамик ферментативного процесса.

Результаты

Результаты: По характеру зависимости обратных кинетических показателей было установлено, что в пределах каждой концентрации ферментного препарата значение асимптотического темпа процесса Vmax = const, а константа Михаэлиса Km непрерывно увеличивается. Для комплексной оценки исследуемых динамик была разработана система критериев, включающая критерий изменчивости константы Михаэлиса Q (Km), дискретный критерий оптимальности Z, комплексный критерий оптимальности декатионизации F и критерий оценки Q, сопровождаемые 4 постулируемыми условиями оптимизации.

Выводы

Выводы: В результате разработанная система критериев позволяет решать сразу три задачи — определение целесообразности предварительной декатионизации, оптимальные концентрации комплексона и гомоферментного препарата.

Introduction

Introduction: The use of enzyme preparations for processing plant raw materials imposes some restrictions determined by the structure of enzyme molecules and the properties of the medium. One of the main limiting factors is the possible presence of an inhibitor in the medium that suppresses catalytic activity. Polyvalent metal cations can be a potential inhibitor, therefore, determining the expediency of decationization of the medium before enzymatic treatment is relevant.

Purpose

Purpose: The aim of the work is to develop an approach to determining the feasibility of preliminary decationization of raw materials before the enzymatic process.

Materials and Methods

Materials and Methods: Non-granulated dry non-massaged beet pulp was used as a substrate. Enzymatic treatment was carried out with a homoenzyme preparation of rhamnogalacturonan lyase action at a concentration of 0.00, 0.05, 0.1, 0.2, 0.4 and 0.8 % by weight of the dry substrate. The pretreatment of the swollen raw materials was carried out by 0.00, 0.20, 0.50 and 0.91 units of 2.87 % aqueous solution of complexon. The response factor was electrical conductivity. The enzymatic process was carried out at a temperature of 45 °C for 8 hours. By approximating the experimental data for each research variant, a mathematical description of the dynamics of the enzymatic process was obtained.

Results

Results: By the nature of the dependence of the inverse kinetic parameters, it was found that within each concentration of the enzyme preparation, the value of the asymptotic rate of the process Vmax = const, and the Michaelis constant Km continuously increases. For a comprehensive assessment of the studied dynamics, a system of criteria was developed, including the criterion of variability of the Michaelis constant Q (Km), the discrete optimality criterion Z, the complex optimality criterion of decationization F and the evaluation criterion Q, accompanied by 4 postulated optimization conditions.

Conclusions

Conclusions: As a result, the developed system of criteria makes it possible to solve three tasks at once – determining the feasibility of pre-decationization, optimal concentrations of complexon and homoenzyme preparation. 

ферментативная активностьфрагментацияпротопектиновый комплексингибированиедекатионизациярастительное сырьёenzymatic activityfragmentationprotopectin complexinhibitiondecationizationvegetable raw materialsВсероссийский научный центр пищевых систем им. В.М. ГорбатоваReferencesRimareva, L. V., Serba, E. M., Sokolova, E. N., Borshcheva, Yu. A., Ignatova, N. I. (2017). Enzyme preparations and biocatalytic processes in the food industry. Voprosy pitaniia = Problems of Nutrition, 86(5): 62–74. https://doi.org/10.24411/0042-8833-2017-00078Rimareva, L. V., Serba, E. M., Sokolova, E. N., Borshcheva, Yu. A., Ignatova, N. I. (2017). Enzyme preparations and biocatalytic processes in the food industry. Voprosy pitaniia = Problems of Nutrition, 86(5): 62–74. (in Russian). https://doi.org/10.24411/0042-8833-2017-00078Mandake, M. B., Das, U., Phad, L., Busamwar, S. (2020). Application of Enzyme Immobilization in the Food Industry. International Journal of Advanced Research in Chemical Science, 7(2): 6–10. http://dx.doi.org/10.20431/2349-0403.0702002Mandake, M. B., Das, U., Phad, L., Busamwar, S. (2020). Application of Enzyme Immobilization in the Food Industry. International Journal of Advanced Research in Chemical Science, 7(2): 6–10. http://dx.doi.org/10.20431/2349-0403.0702002Ochiai, A., Itoh, T., Kawamata, A., Hashimoto, W., Murata, K. (2007). Plant cell wall degradation by saprophytic Bacillus subtilis strains: gene clusters responsible for rhamnogalacturonan depolymerization. Applied and Environmental Microbiology, 73(12): 3803–3813. https://doi.org/10.1128/aem.00147-07Ochiai, A., Itoh, T., Kawamata, A., Hashimoto, W., Murata, K. (2007). Plant cell wall degradation by saprophytic Bacillus subtilis strains: gene clusters responsible for rhamnogalacturonan depolymerization. Applied and Environmental Microbiology, 73(12): 3803–3813. https://doi.org/10.1128/aem.00147-07Silva, I. R., Larsen, D. M., Meyer, A. S., Mikkelsen, J. D. (2011). Identification, expression, and characterization of a novel bacterial RGI lyase enzyme for the production of bio-functional fibers. Enzyme and Microbial Technology, 49(2): 160–166. https://doi.org/10.1016/j.enzmictec.2011.04.015Silva, I. R., Larsen, D. M., Meyer, A. S., Mikkelsen, J. D. (2011). Identification, expression, and characterization of a novel bacterial RGI lyase enzyme for the production of bio-functional fibers. Enzyme and Microbial Technology, 49(2): 160–166. https://doi.org/10.1016/j.enzmictec.2011.04.015Dhillon, A., Fernandes, V. O., Dias, F. M. V., Prates, J. A. M., Ferreira, L. M. A., Fontes, C. M. G. A., Centeno, M. S., Goyal, A. (2016). A New Member of Family 11 Polysaccharide Lyase, Rhamnogalacturonan Lyase (CtRGLf) from Clostridium thermocellum. Molecular Biotechnology, 58: 232–240. https://doi.org/10.1007/s12033-016-9921-6Dhillon, A., Fernandes, V. O., Dias, F. M. V., Prates, J. A. M., Ferreira, L. M. A., Fontes, C. M. G. A., Centeno, M. S., Goyal, A. (2016). A New Member of Family 11 Polysaccharide Lyase, Rhamnogalacturonan Lyase (CtRGLf) from Clostridium thermocellum. Molecular Biotechnology, 58: 232–240. https://doi.org/10.1007/s12033-016-9921-6Yuan, Y., Zhang, X.-Y., Zhao, Y., Zhang, H., Zhou, Y.-F., Gao, J. (2019). A Novel PL9 Pectate Lyase from Paenibacillus polymyxa KF-1: Cloning, Expression, and Its Application in Pectin Degradation. International Journal of Molecular Sciences, 20(12): 3060. https://doi.org/10.3390/ijms20123060Yuan, Y., Zhang, X.-Y., Zhao, Y., Zhang, H., Zhou, Y.-F., Gao, J. (2019). A Novel PL9 Pectate Lyase from Paenibacillus polymyxa KF-1: Cloning, Expression, and Its Application in Pectin Degradation. International Journal of Molecular Sciences, 20(12): 3060. https://doi.org/10.3390/ijms20123060Wang, W., Wang, Y., Yi, H., Liu, Y., Zhang, G., Zhang, L., Mayo, K.H., Yuan, Y., Zhou, Y. (2022). Biochemical Characterization of Two Rhamnogalacturonan Lyases from Bacteroides ovatus ATCC 8483 with Preference for RG-I Substrates. Frontiers in Microbiology, 12: 799875. https://doi.org/10.3389/fmicb.2021.799875Wang, W., Wang, Y., Yi, H., Liu, Y., Zhang, G., Zhang, L., Mayo, K.H., Yuan, Y., Zhou, Y. (2022). Biochemical Characterization of Two Rhamnogalacturonan Lyases from Bacteroides ovatus ATCC 8483 with Preference for RG-I Substrates. Frontiers in Microbiology, 12: 799875. https://doi.org/10.3389/fmicb.2021.799875Jensen, M. H., Otten, H., Christensen, U., Borchert, T. V., Christensen, L. L. H., Larsen, S., Leggi, L. L. (2010). Structural and Biochemical Studies Elucidate the Mechanism of Rhamnogalacturonan Lyase from Aspergillus aculeatus. Journal of Molecular Biology, 404(1): 100–111. https://doi.org/10.1016/j.jmb.2010.09.013Jensen, M. H., Otten, H., Christensen, U., Borchert, T. V., Christensen, L. L. H., Larsen, S., Leggi, L. L. (2010). Structural and Biochemical Studies Elucidate the Mechanism of Rhamnogalacturonan Lyase from Aspergillus aculeatus. Journal of Molecular Biology, 404(1): 100–111. https://doi.org/10.1016/j.jmb.2010.09.013Laatu, M, Condemine, G. (2003). Rhamnogalacturonate lyase RhiE is secreted by the out system in Erwinia chrysanthemi. Journal of Bacteriology, 185(5): 1642–1649. https://doi.org/10.1128/JB.185.5.1642-1649.2003Laatu, M, Condemine, G. (2003). Rhamnogalacturonate lyase RhiE is secreted by the out system in Erwinia chrysanthemi. Journal of Bacteriology, 185(5): 1642–1649. https://doi.org/10.1128/JB.185.5.1642-1649.2003Iwai, M., Yamada, H., Ikemoto, T., Matsumoto, S., Fujiwara, D., Takenaka, S., Sakamoto, T. (2015). Biochemical Characterization and Overexpression of an Endo-rhamnogalacturonan Lyase from Penicillium chrysogenum. Molecular Biotechnology, 57(6): 539–548. https://doi.org/10.1007/s12033-015-9847-4Iwai, M., Yamada, H., Ikemoto, T., Matsumoto, S., Fujiwara, D., Takenaka, S., Sakamoto, T. (2015). Biochemical Characterization and Overexpression of an Endo-rhamnogalacturonan Lyase from Penicillium chrysogenum. Molecular Biotechnology, 57(6): 539–548. https://doi.org/10.1007/s12033-015-9847-4Morales-Quintana, L., Ramos, P., Méndez-Yáñez, A. (2022). Rhamnogalacturonan Endolyase Family 4 Enzymes: An Update on Their Importance in the Fruit Ripening Process. Horticulturae, 8: 465. https://doi.org/10.3390/horticulturae8050465Morales-Quintana, L., Ramos, P., Méndez-Yáñez, A. (2022). Rhamnogalacturonan Endolyase Family 4 Enzymes: An Update on Their Importance in the Fruit Ripening Process. Horticulturae, 8: 465. https://doi.org/10.3390/horticulturae8050465Fujimoto, Z., Jackson, A., Michikawa, M., Maehara, T., Momma, M., Henrissat, B., Gilbert, H. J., Kaneko, S. (2013). The structure of a Streptomyces avermitilis α-L-rhamnosidase reveals a novel carbohydrate-binding module CBM67 within the six-domain arrangement. The Journal of Biological Chemistry, 288(17): 12376–12385. https://doi.org/10.1074/jbc.M113.460097Fujimoto, Z., Jackson, A., Michikawa, M., Maehara, T., Momma, M., Henrissat, B., Gilbert, H. J., Kaneko, S. (2013). The structure of a Streptomyces avermitilis α-L-rhamnosidase reveals a novel carbohydrate-binding module CBM67 within the six-domain arrangement. The Journal of Biological Chemistry, 288(17): 12376–12385. https://doi.org/10.1074/jbc.M113.460097O'Neill, E. C., Stevenson, C. E., Paterson, M. J., Rejzek, M., Chauvin, A. L., Lawson, D. M., Field, R. A. (2015). Crystal structure of a novel two domain GH78 family α-rhamnosidase from Klebsiella oxytoca with rhamnose bound. Protein, 83(9): 1742–1749. https://doi.org/10.1002/prot.24807O'Neill, E. C., Stevenson, C. E., Paterson, M. J., Rejzek, M., Chauvin, A. L., Lawson, D. M., Field, R. A. (2015). Crystal structure of a novel two domain GH78 family α-rhamnosidase from Klebsiella oxytoca with rhamnose bound. Protein, 83(9): 1742–1749. https://doi.org/10.1002/prot.24807Furusawa, G., Azami, N. A., The, A.-H. (2021). Genes for degradation and utilization of uronic acid-containing polysaccharides of a marine bacterium Catenovulum sp. CCB-QB4. PeerJ, 9: 10929. http://doi.org/10.7717/peerj.10929Furusawa, G., Azami, N. A., The, A.-H. (2021). Genes for degradation and utilization of uronic acid-containing polysaccharides of a marine bacterium Catenovulum sp. CCB-QB4. PeerJ, 9: 10929. http://doi.org/10.7717/peerj.10929Robin, T., Reuveni, S., Urbakh, M. (2018). Single-molecule theory of enzymatic inhibition. Nature Communications, 9, 779. https://doi.org/10.1038/s41467-018-02995-6Robin, T., Reuveni, S., Urbakh, M. (2018). Single-molecule theory of enzymatic inhibition. Nature Communications, 9: 779. https://doi.org/10.1038/s41467-018-02995-6Ivanova, Yu. V. (2022). Enzymes seen by restorers eyes or the ingenious gentleman don Quixote of la Mancha. Art Heritage. Research. Storage. Conservation, 4: 7–20. (in Russ)Ivanova, Yu. V. (2022). Enzymes seen by restorers eyes or the ingenious gentleman don Quixote of la Mancha. Art Heritage. Research. Storage. Conservation, 4: 7–20. (in Russian).Patadiya, N., Panchal, N., Vaghela, V. (2021). A Review on Enzyme Inhibitors. International Research Journal of Pharmacy, 12(6): 60–66. http://dx.doi.org/10.7897/2230-8407.1206145Patadiya, N., Panchal, N., Vaghela, V. (2021). A Review on Enzyme Inhibitors. International Research Journal of Pharmacy, 12(6): 60–66. http://dx.doi.org/10.7897/2230-8407.1206145Zhang, Y., Ren, H., Wang, Y., Chen, K., Fang, B., Wang, Sh. (2016). Bioinspired Immobilization of Glycerol Dehydrogenase by Metal Ion-Chelated Polyethyleneimines as Artificial Polypeptides. Scientific Reports, 6, 24163. https://doi.org/10.1038/srep24163Zhang, Y., Ren, H., Wang, Y., Chen, K., Fang, B., Wang, Sh. (2016). Bioinspired Immobilization of Glycerol Dehydrogenase by Metal Ion-Chelated Polyethyleneimines as Artificial Polypeptides. Scientific Reports, 6: 24163. https://doi.org/10.1038/srep24163Joslyn, M. A. (1963). The Chemistry of Protopectin: A Critical Review of Historical Data and Recent Developments. Advances in Food Research, 11: 1–107. https://doi.org/10.1016/S0065-2628(08)60064-6Joslyn, M. A. (1963). The Chemistry of Protopectin: A Critical Review of Historical Data and Recent Developments. Advances in Food Research, 11: 1–107. https://doi.org/10.1016/S0065-2628(08)60064-6Kondratenko, V. V., Petrov, A. N., Kondratenko T. Yu. (2022). Basic notions for kinetics of enzymatic processes and approaches to its determination. Vsyo o myase, 6: 12–19. https://doi.org/10.21323/2071-2499-2022-6-12-19 (in Russ)Kondratenko, V. V., Petrov, A. N., Kondratenko T. Yu. (2022). Basic notions for kinetics of enzymatic processes and approaches to its determination. Vsyo o myase, 6: 12–19. (in Russian). https://doi.org/10.21323/2071-2499-2022-6-12-19Kondratenko, V. V., Tsaryova, M. A., Kondratenko, T. Yu., Davydova, A. Yu., Alabina, N. M. (2018). O dekationizatsii pektinosoderzhashchego syrʹya na primere sveklovichnogo zhoma [Decation of pectin-containing raw materials using beet pulp as an example]. Nauchnye trudy Severo-Kavkazskogo Federalʹnogo nauchnogo tsentra sadovodstva, vinogradarstva i vinodeliya [Scientific Proceedings of the North Caucasus Federal Scientific Center for Horticulture, Viticulture and Winemaking], 21: 42–48. (in Russ)Kondratenko, V. V., Tsaryova, M. A., Kondratenko, T. Yu., Davydova, A. Yu., Alabina, N. M. (2018). O dekationizatsii pektinosoderzhashchego syrʹya na primere sveklovichnogo zhoma [Decation of pectin-containing raw materials using beet pulp as an example]. Nauchnye trudy Severo-Kavkazskogo Federalʹnogo nauchnogo tsentra sadovodstva, vinogradarstva i vinodeliya [Scientific Proceedings of the North Caucasus Federal Scientific Center for Horticulture, Viticulture and Winemaking], 21: 42–48. (in Russian).Seltman, Y.J. (2018). Experimental Design and Analysis. Pittsburgh: Carnegie Mellon University. 414 p. https://www.stat.cmu.edu/~hseltman/309/Book/Book.pdf.Seltman, Y.J. (2018). Experimental Design and Analysis. Pittsburgh: Carnegie Mellon University 414 p. https://www.stat.cmu.edu/~hseltman/309/Book/Book.pdf.Kondratenko, V. V., Petrov, A. N., Kondratenko, T. Yu. (2022). Razrabotka metodov opredeleniya optimalʹnoy kontsentratsii fermentov na osnovanii analiza dinamiki fermentirovaniya [Development the methods for determining the optimal concentration of enzymes based on analysis of fermentation dynamics]. Pishchevaya promyshlennostʹ [Food Industry], 11: 67–74. https://doi.org/10.52653/PPI.2022.11.11.016 (in Russian).Kondratenko, V. V., Petrov, A. N., Kondratenko, T. Yu. (2022). Razrabotka metodov opredeleniya optimalʹnoy kontsentratsii fermentov na osnovanii analiza dinamiki fermentirovaniya [Development the methods for determining the optimal concentration of enzymes based on analysis of fermentation dynamics]. Pishchevaya promyshlennostʹ [Food Industry], 11: 67–74. (in Russian). https://doi.org/10.52653/PPI.2022.11.11.016

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