Development of a Dual-Stage CIM<sup>®</sup> CDI Reactor with Immobilized Glucuronan Lyases and Laccases for Sustainable Synthesis of Antioxidant Phenolized Oligoglucuronan

Immobilized enzyme reactors (IMERs) are critical tools for developing novel oligosaccharides based on the enzymatic catalysis of polysaccharides. In this paper, a novel glucuronan lyase from <i>Peteryoungia rosettiformans</i> was produced, purified, and then immobilized on a CIM<sup&g...

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Main Authors: Xiaoyang Hou, Pascal Dubessay, Gwendoline Christophe, Nicolas Bridiau, Pierre-Edouard Bodet, Mounir Traikia, Mugilan Damadoran Raja, Thierry Maugard, Aleš Štrancar, Fabrice Audonnet, Philippe Michaud, Guillaume Pierre
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Language:English
Published: MDPI AG 2024-11-01
Series:Polysaccharides
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Online Access:https://www.mdpi.com/2673-4176/5/4/47
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author Xiaoyang Hou
Pascal Dubessay
Gwendoline Christophe
Nicolas Bridiau
Pierre-Edouard Bodet
Mounir Traikia
Mugilan Damadoran Raja
Thierry Maugard
Aleš Štrancar
Fabrice Audonnet
Philippe Michaud
Guillaume Pierre
author_facet Xiaoyang Hou
Pascal Dubessay
Gwendoline Christophe
Nicolas Bridiau
Pierre-Edouard Bodet
Mounir Traikia
Mugilan Damadoran Raja
Thierry Maugard
Aleš Štrancar
Fabrice Audonnet
Philippe Michaud
Guillaume Pierre
author_sort Xiaoyang Hou
collection DOAJ
description Immobilized enzyme reactors (IMERs) are critical tools for developing novel oligosaccharides based on the enzymatic catalysis of polysaccharides. In this paper, a novel glucuronan lyase from <i>Peteryoungia rosettiformans</i> was produced, purified, and then immobilized on a CIM<sup>®</sup> CDI disk for cleaving glucuronan. The results showed that around 63.6% of glycuronan lyases (800.9 μg) were immobilized on the disk. The V<sub>max</sub> values of immobilized glucuronan lyases did not significantly change (56.9 ± 4.7 μM∙min<sup>−1</sup>), while the K<sub>m</sub> values (0.310 ± 0.075 g∙L<sup>−1</sup>) increased by 2.5 times. It is worth noting that immobilized glucuronan lyases overcame the catalytic inhibition of free enzymes observed under high glucuronan concentrations (0.5–2 g∙L<sup>−1</sup>). circumscribed central composite design (CCCD) and response surface methodology (RSM) showed that glucuronan concentration, flow rate, and reaction time significantly affected the yield of oligoglucuronans. The degree of polymerization (DP) of degraded glucuronan ranged from DP 2–8 according to the results obtained by high performance anion exchange chromatography coupled with a pulsed amperometric detector (HPAEC-PAD). The IMER retained 50.9% activity after running 2373 column volumes of glucuronan. Finally, this glucuronan lyase reactor was tentatively connected to an immobilized laccase reactor to depolymerize, and gallic acid (GA) was added to glucuronan. Approximately 8.5 mg of GA was added onto 1 g of initial glucuronan, and the GA–oligoglucuronan conjugates showed notable antioxidant activity.
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spelling doaj-art-ca1f63bf95c34d4c8a1e6a50ec52f1cc2025-08-20T02:01:09ZengMDPI AGPolysaccharides2673-41762024-11-015474376010.3390/polysaccharides5040047Development of a Dual-Stage CIM<sup>®</sup> CDI Reactor with Immobilized Glucuronan Lyases and Laccases for Sustainable Synthesis of Antioxidant Phenolized OligoglucuronanXiaoyang Hou0Pascal Dubessay1Gwendoline Christophe2Nicolas Bridiau3Pierre-Edouard Bodet4Mounir Traikia5Mugilan Damadoran Raja6Thierry Maugard7Aleš Štrancar8Fabrice Audonnet9Philippe Michaud10Guillaume Pierre11Clermont Auvergne INP, CNRS, Institut Pascal, Université Clermont Auvergne, F-63000 Clermont-Ferrand, FranceClermont Auvergne INP, CNRS, Institut Pascal, Université Clermont Auvergne, F-63000 Clermont-Ferrand, FranceClermont Auvergne INP, CNRS, Institut Pascal, Université Clermont Auvergne, F-63000 Clermont-Ferrand, FranceUMR CNRS 7266 LIENSs, La Rochelle Université, F-17000 La Rochelle, FranceUMR CNRS 7266 LIENSs, La Rochelle Université, F-17000 La Rochelle, FranceClermont Auvergne INP, CNRS, Institut de Chimie de Clermont-Ferrand, Université Clermont Auvergne, F-63000 Clermont-Ferrand, FranceSchool of Bio Sciences and Technology, Vellore Institute of Technology (VIT) University, Vellore 632014, Tamil Nadu, IndiaUMR CNRS 7266 LIENSs, La Rochelle Université, F-17000 La Rochelle, FranceSartorius BIA Separations d.o.o., Mirce 21, SI-5270 Ajdovščina, SloveniaClermont Auvergne INP, CNRS, Institut Pascal, Université Clermont Auvergne, F-63000 Clermont-Ferrand, FranceClermont Auvergne INP, CNRS, Institut Pascal, Université Clermont Auvergne, F-63000 Clermont-Ferrand, FranceClermont Auvergne INP, CNRS, Institut Pascal, Université Clermont Auvergne, F-63000 Clermont-Ferrand, FranceImmobilized enzyme reactors (IMERs) are critical tools for developing novel oligosaccharides based on the enzymatic catalysis of polysaccharides. In this paper, a novel glucuronan lyase from <i>Peteryoungia rosettiformans</i> was produced, purified, and then immobilized on a CIM<sup>®</sup> CDI disk for cleaving glucuronan. The results showed that around 63.6% of glycuronan lyases (800.9 μg) were immobilized on the disk. The V<sub>max</sub> values of immobilized glucuronan lyases did not significantly change (56.9 ± 4.7 μM∙min<sup>−1</sup>), while the K<sub>m</sub> values (0.310 ± 0.075 g∙L<sup>−1</sup>) increased by 2.5 times. It is worth noting that immobilized glucuronan lyases overcame the catalytic inhibition of free enzymes observed under high glucuronan concentrations (0.5–2 g∙L<sup>−1</sup>). circumscribed central composite design (CCCD) and response surface methodology (RSM) showed that glucuronan concentration, flow rate, and reaction time significantly affected the yield of oligoglucuronans. The degree of polymerization (DP) of degraded glucuronan ranged from DP 2–8 according to the results obtained by high performance anion exchange chromatography coupled with a pulsed amperometric detector (HPAEC-PAD). The IMER retained 50.9% activity after running 2373 column volumes of glucuronan. Finally, this glucuronan lyase reactor was tentatively connected to an immobilized laccase reactor to depolymerize, and gallic acid (GA) was added to glucuronan. Approximately 8.5 mg of GA was added onto 1 g of initial glucuronan, and the GA–oligoglucuronan conjugates showed notable antioxidant activity.https://www.mdpi.com/2673-4176/5/4/47glucuronanglucuronan lyaseoligoglucuronangallic acidCIM<sup>®</sup> CDI diskimmobilized enzyme reactors
spellingShingle Xiaoyang Hou
Pascal Dubessay
Gwendoline Christophe
Nicolas Bridiau
Pierre-Edouard Bodet
Mounir Traikia
Mugilan Damadoran Raja
Thierry Maugard
Aleš Štrancar
Fabrice Audonnet
Philippe Michaud
Guillaume Pierre
Development of a Dual-Stage CIM<sup>®</sup> CDI Reactor with Immobilized Glucuronan Lyases and Laccases for Sustainable Synthesis of Antioxidant Phenolized Oligoglucuronan
Polysaccharides
glucuronan
glucuronan lyase
oligoglucuronan
gallic acid
CIM<sup>®</sup> CDI disk
immobilized enzyme reactors
title Development of a Dual-Stage CIM<sup>®</sup> CDI Reactor with Immobilized Glucuronan Lyases and Laccases for Sustainable Synthesis of Antioxidant Phenolized Oligoglucuronan
title_full Development of a Dual-Stage CIM<sup>®</sup> CDI Reactor with Immobilized Glucuronan Lyases and Laccases for Sustainable Synthesis of Antioxidant Phenolized Oligoglucuronan
title_fullStr Development of a Dual-Stage CIM<sup>®</sup> CDI Reactor with Immobilized Glucuronan Lyases and Laccases for Sustainable Synthesis of Antioxidant Phenolized Oligoglucuronan
title_full_unstemmed Development of a Dual-Stage CIM<sup>®</sup> CDI Reactor with Immobilized Glucuronan Lyases and Laccases for Sustainable Synthesis of Antioxidant Phenolized Oligoglucuronan
title_short Development of a Dual-Stage CIM<sup>®</sup> CDI Reactor with Immobilized Glucuronan Lyases and Laccases for Sustainable Synthesis of Antioxidant Phenolized Oligoglucuronan
title_sort development of a dual stage cim sup r sup cdi reactor with immobilized glucuronan lyases and laccases for sustainable synthesis of antioxidant phenolized oligoglucuronan
topic glucuronan
glucuronan lyase
oligoglucuronan
gallic acid
CIM<sup>®</sup> CDI disk
immobilized enzyme reactors
url https://www.mdpi.com/2673-4176/5/4/47
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