Efficient Hydrolysis of Sugar Beet Pulp Using Novel Enzyme Complexes

Sugar beet pulp is a byproduct of white sugar production, and it is quite significant in terms of volume. Every year, tens of millions of tons of beet pulp are produced around the world. However, only a fraction of it is currently used, mainly as animal feed. The composition of beet pulp includes pl...

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Main Authors: Maria I. Komarova, Margarita V. Semenova, Pavel V. Volkov, Igor A. Shashkov, Alexandra M. Rozhkova, Ivan N. Zorov, Sergei A. Kurzeev, Aidar D. Satrutdinov, Ekaterina A. Rubtsova, Arkady P. Sinitsyn
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Language:English
Published: MDPI AG 2025-01-01
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Online Access:https://www.mdpi.com/2073-4395/15/1/101
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author Maria I. Komarova
Margarita V. Semenova
Pavel V. Volkov
Igor A. Shashkov
Alexandra M. Rozhkova
Ivan N. Zorov
Sergei A. Kurzeev
Aidar D. Satrutdinov
Ekaterina A. Rubtsova
Arkady P. Sinitsyn
author_facet Maria I. Komarova
Margarita V. Semenova
Pavel V. Volkov
Igor A. Shashkov
Alexandra M. Rozhkova
Ivan N. Zorov
Sergei A. Kurzeev
Aidar D. Satrutdinov
Ekaterina A. Rubtsova
Arkady P. Sinitsyn
author_sort Maria I. Komarova
collection DOAJ
description Sugar beet pulp is a byproduct of white sugar production, and it is quite significant in terms of volume. Every year, tens of millions of tons of beet pulp are produced around the world. However, only a fraction of it is currently used, mainly as animal feed. The composition of beet pulp includes plant polysaccharides, such as cellulose, arabinan, and pectin. Through the process of enzymatic hydrolysis, these polysaccharides are converted into technical C6/C5 sugars, which can be further used as a substrate for the microbial synthesis of various substances, including biofuels, organic acids, and other green chemistry molecules. The current study was designed with a primary objective that focused on the development of a strain that had the potential for enhanced productivity and the capacity to produce enzymes suitable for beet pulp hydrolysis. The <i>pelA</i> and <i>abfA</i> genes, which encode pectin lyase and arabinofuranosidase, respectively, in the fungus <i>Penicillium canescens</i> (VKPM F-178), were cloned and successfully expressed in the recipient strain <i>Penicillium verruculosum</i> B1-537 (VKPM F-3972D). New recombinant strains were created using the expression system of the mycelial fungus <i>P. verruculosum</i> B1-537, which is capable of simultaneously producing pectin lyase and arabinofuranosidase, as well as homologous cellulases. The screening of strains for increased enzymatic activity towards citrus pectin, sugar beet branched arabinan, and microcrystalline cellulose revealed that a B4 clone of <i>P. verruculosum</i> exhibited the greatest potential in sugar beet pulp cake hydrolysis. This clone was selected as the basis for the creation of a new enzyme preparation with enhanced pectin lyase, arabinase, and cellulase activities. The component composition of the enzyme preparation was determined, and the results indicated that the enzyme content comprised approximately 11% pectin lyase, 40% arabinofuranosidase, and 40% cellulases. The primary products of the enzymatic hydrolysis of the unpretreated beet pulp cake were arabinose and glucose. The degree of arabinan and cellulose conversion was observed to be up to 50% and 80%, respectively, after a period of 48 to 72 h of hydrolysis. The new B4 preparation was observed to be highly efficacious in the hydrolysis of beet cake at elevated concentrations of solids (up to 300 g/L) within the reaction mixture. The newly developed strain, as a producer of pectin lyase, arabinofuranosidase, and cellulase complexes, has the potential to be utilized for the bioconversion of sugar beet processing wastes and for the efficient generation of highly concentrated solutions of technical sugars for further implementation in processes of microbial synthesis.
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spelling doaj-art-0ff1c844468a4a7fb8a82ac0cf7a9c2d2025-01-24T13:16:43ZengMDPI AGAgronomy2073-43952025-01-0115110110.3390/agronomy15010101Efficient Hydrolysis of Sugar Beet Pulp Using Novel Enzyme ComplexesMaria I. Komarova0Margarita V. Semenova1Pavel V. Volkov2Igor A. Shashkov3Alexandra M. Rozhkova4Ivan N. Zorov5Sergei A. Kurzeev6Aidar D. Satrutdinov7Ekaterina A. Rubtsova8Arkady P. Sinitsyn9Department of Chemistry, Lomonosov Moscow State University, 119991 Moscow, RussiaFederal Research Centre ‘Fundamental of Biotechnology’ of the Russian Academy of Sciences (FRC Biotechnology RAS), 119071 Moscow, RussiaFederal Research Centre ‘Fundamental of Biotechnology’ of the Russian Academy of Sciences (FRC Biotechnology RAS), 119071 Moscow, RussiaFederal Research Centre ‘Fundamental of Biotechnology’ of the Russian Academy of Sciences (FRC Biotechnology RAS), 119071 Moscow, RussiaFederal Research Centre ‘Fundamental of Biotechnology’ of the Russian Academy of Sciences (FRC Biotechnology RAS), 119071 Moscow, RussiaDepartment of Chemistry, Lomonosov Moscow State University, 119991 Moscow, RussiaDepartment of Chemistry, Lomonosov Moscow State University, 119991 Moscow, RussiaFederal Research Centre ‘Fundamental of Biotechnology’ of the Russian Academy of Sciences (FRC Biotechnology RAS), 119071 Moscow, RussiaFederal Research Centre ‘Fundamental of Biotechnology’ of the Russian Academy of Sciences (FRC Biotechnology RAS), 119071 Moscow, RussiaDepartment of Chemistry, Lomonosov Moscow State University, 119991 Moscow, RussiaSugar beet pulp is a byproduct of white sugar production, and it is quite significant in terms of volume. Every year, tens of millions of tons of beet pulp are produced around the world. However, only a fraction of it is currently used, mainly as animal feed. The composition of beet pulp includes plant polysaccharides, such as cellulose, arabinan, and pectin. Through the process of enzymatic hydrolysis, these polysaccharides are converted into technical C6/C5 sugars, which can be further used as a substrate for the microbial synthesis of various substances, including biofuels, organic acids, and other green chemistry molecules. The current study was designed with a primary objective that focused on the development of a strain that had the potential for enhanced productivity and the capacity to produce enzymes suitable for beet pulp hydrolysis. The <i>pelA</i> and <i>abfA</i> genes, which encode pectin lyase and arabinofuranosidase, respectively, in the fungus <i>Penicillium canescens</i> (VKPM F-178), were cloned and successfully expressed in the recipient strain <i>Penicillium verruculosum</i> B1-537 (VKPM F-3972D). New recombinant strains were created using the expression system of the mycelial fungus <i>P. verruculosum</i> B1-537, which is capable of simultaneously producing pectin lyase and arabinofuranosidase, as well as homologous cellulases. The screening of strains for increased enzymatic activity towards citrus pectin, sugar beet branched arabinan, and microcrystalline cellulose revealed that a B4 clone of <i>P. verruculosum</i> exhibited the greatest potential in sugar beet pulp cake hydrolysis. This clone was selected as the basis for the creation of a new enzyme preparation with enhanced pectin lyase, arabinase, and cellulase activities. The component composition of the enzyme preparation was determined, and the results indicated that the enzyme content comprised approximately 11% pectin lyase, 40% arabinofuranosidase, and 40% cellulases. The primary products of the enzymatic hydrolysis of the unpretreated beet pulp cake were arabinose and glucose. The degree of arabinan and cellulose conversion was observed to be up to 50% and 80%, respectively, after a period of 48 to 72 h of hydrolysis. The new B4 preparation was observed to be highly efficacious in the hydrolysis of beet cake at elevated concentrations of solids (up to 300 g/L) within the reaction mixture. The newly developed strain, as a producer of pectin lyase, arabinofuranosidase, and cellulase complexes, has the potential to be utilized for the bioconversion of sugar beet processing wastes and for the efficient generation of highly concentrated solutions of technical sugars for further implementation in processes of microbial synthesis.https://www.mdpi.com/2073-4395/15/1/101beet pulpplant polysaccharidesenzymatic conversion<i>Penicillium verruculosum</i>
spellingShingle Maria I. Komarova
Margarita V. Semenova
Pavel V. Volkov
Igor A. Shashkov
Alexandra M. Rozhkova
Ivan N. Zorov
Sergei A. Kurzeev
Aidar D. Satrutdinov
Ekaterina A. Rubtsova
Arkady P. Sinitsyn
Efficient Hydrolysis of Sugar Beet Pulp Using Novel Enzyme Complexes
Agronomy
beet pulp
plant polysaccharides
enzymatic conversion
<i>Penicillium verruculosum</i>
title Efficient Hydrolysis of Sugar Beet Pulp Using Novel Enzyme Complexes
title_full Efficient Hydrolysis of Sugar Beet Pulp Using Novel Enzyme Complexes
title_fullStr Efficient Hydrolysis of Sugar Beet Pulp Using Novel Enzyme Complexes
title_full_unstemmed Efficient Hydrolysis of Sugar Beet Pulp Using Novel Enzyme Complexes
title_short Efficient Hydrolysis of Sugar Beet Pulp Using Novel Enzyme Complexes
title_sort efficient hydrolysis of sugar beet pulp using novel enzyme complexes
topic beet pulp
plant polysaccharides
enzymatic conversion
<i>Penicillium verruculosum</i>
url https://www.mdpi.com/2073-4395/15/1/101
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