Immobilized Crosslinked Pectinase Preparation on Porous ZSM-5 Zeolites as Reusable Biocatalysts for Ultra-Efficient Hydrolysis of β-Glycosidic Bonds
In this study, we immobilized pectinase preparation on porous zeolite ZSM-5 as an enzyme carrier. We realized this immobilized enzyme catalyst, pectinase preparation@ZSM-5, via a simple combined strategy involving the van der Waals adsorption of pectinase preparation followed by crosslinking of the...
Main Authors: | , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Frontiers Media S.A.
2021-08-01
|
Series: | Frontiers in Chemistry |
Subjects: | |
Online Access: | https://www.frontiersin.org/articles/10.3389/fchem.2021.677868/full |
id |
doaj-cd06cbd3d5ef4a67b65032d7f7b08a2f |
---|---|
record_format |
Article |
spelling |
doaj-cd06cbd3d5ef4a67b65032d7f7b08a2f2021-08-11T07:35:20ZengFrontiers Media S.A.Frontiers in Chemistry2296-26462021-08-01910.3389/fchem.2021.677868677868Immobilized Crosslinked Pectinase Preparation on Porous ZSM-5 Zeolites as Reusable Biocatalysts for Ultra-Efficient Hydrolysis of β-Glycosidic BondsCan Liu0Liming Zhang1Li Tan2Yueping Liu3Weiqian Tian4Lanqing Ma5Key Laboratory for Northern Urban Agriculture of Ministry of Agriculture and Rural Affairs, Beijing University of Agriculture, Beijing, ChinaDepartment of Fibre and Polymer Technology, KTH Royal Institute of Technology, Stockholm, SwedenKey Laboratory for Northern Urban Agriculture of Ministry of Agriculture and Rural Affairs, Beijing University of Agriculture, Beijing, ChinaKey Laboratory for Northern Urban Agriculture of Ministry of Agriculture and Rural Affairs, Beijing University of Agriculture, Beijing, ChinaDepartment of Fibre and Polymer Technology, KTH Royal Institute of Technology, Stockholm, SwedenKey Laboratory for Northern Urban Agriculture of Ministry of Agriculture and Rural Affairs, Beijing University of Agriculture, Beijing, ChinaIn this study, we immobilized pectinase preparation on porous zeolite ZSM-5 as an enzyme carrier. We realized this immobilized enzyme catalyst, pectinase preparation@ZSM-5, via a simple combined strategy involving the van der Waals adsorption of pectinase preparation followed by crosslinking of the adsorbed pectinase preparation with glutaraldehyde over ZSM-5. Conformal pectinase preparation coverage of various ZSM-5 supports was achieved for the as-prepared pectinase preparation@ZSM-5. The porous pectinase preparation@ZSM-5 catalyst exhibited ultra-efficient biocatalytic activity for hydrolyzing the β-glycosidic bonds in the model substrate 4-nitrophenyl β-D-glucopyranoside, with a broad operating temperature range, high thermal stability, and excellent reusability. The relative activity of pectinase preparation@ZSM-5 at a high temperature (70 °C) was nine times higher than that of free pectinase preparation. Using thermal inactivation kinetic analysis based on the Arrhenius law, pectinase preparation@ZSM-5 showed higher activation energy for denaturation (315 kJ mol−1) and a longer half-life (62 min−1) than free pectinase preparation. Moreover, a Michaelis–Menten enzyme kinetic analysis indicated a higher maximal reaction velocity for pectinase preparation@ZSM-5 (0.22 µmol mg−1 min−1). This enhanced reactivity was attributed to the microstructure of the immobilized pectinase preparation@ZSM-5, which offered a heterogeneous reaction system that decreased the substrate–pectinase preparation binding affinity and modulated the kinetic characteristics of the enzyme. Additionally, pectinase preparation@ZSM-5 showed the best ethanol tolerance among all the reported pectinase preparation-immobilized catalysts, and an activity 247% higher than that of free pectinase preparation at a 10% (v/v) ethanol concentration was measured. Furthermore, pectinase preparation@ZSM-5 exhibited potential for practical engineering applications, promoting the hydrolysis of β-glycosidic bonds in baicalin to convert it into baicalein. This was achieved with a 98% conversion rate, i.e., 320% higher than that of the free enzyme.https://www.frontiersin.org/articles/10.3389/fchem.2021.677868/fullimmobilizationpectinaseβ-glycosidic bondZSM-5 zeoliteheat resistanceethanol tolerance |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Can Liu Liming Zhang Li Tan Yueping Liu Weiqian Tian Lanqing Ma |
spellingShingle |
Can Liu Liming Zhang Li Tan Yueping Liu Weiqian Tian Lanqing Ma Immobilized Crosslinked Pectinase Preparation on Porous ZSM-5 Zeolites as Reusable Biocatalysts for Ultra-Efficient Hydrolysis of β-Glycosidic Bonds Frontiers in Chemistry immobilization pectinase β-glycosidic bond ZSM-5 zeolite heat resistance ethanol tolerance |
author_facet |
Can Liu Liming Zhang Li Tan Yueping Liu Weiqian Tian Lanqing Ma |
author_sort |
Can Liu |
title |
Immobilized Crosslinked Pectinase Preparation on Porous ZSM-5 Zeolites as Reusable Biocatalysts for Ultra-Efficient Hydrolysis of β-Glycosidic Bonds |
title_short |
Immobilized Crosslinked Pectinase Preparation on Porous ZSM-5 Zeolites as Reusable Biocatalysts for Ultra-Efficient Hydrolysis of β-Glycosidic Bonds |
title_full |
Immobilized Crosslinked Pectinase Preparation on Porous ZSM-5 Zeolites as Reusable Biocatalysts for Ultra-Efficient Hydrolysis of β-Glycosidic Bonds |
title_fullStr |
Immobilized Crosslinked Pectinase Preparation on Porous ZSM-5 Zeolites as Reusable Biocatalysts for Ultra-Efficient Hydrolysis of β-Glycosidic Bonds |
title_full_unstemmed |
Immobilized Crosslinked Pectinase Preparation on Porous ZSM-5 Zeolites as Reusable Biocatalysts for Ultra-Efficient Hydrolysis of β-Glycosidic Bonds |
title_sort |
immobilized crosslinked pectinase preparation on porous zsm-5 zeolites as reusable biocatalysts for ultra-efficient hydrolysis of β-glycosidic bonds |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Chemistry |
issn |
2296-2646 |
publishDate |
2021-08-01 |
description |
In this study, we immobilized pectinase preparation on porous zeolite ZSM-5 as an enzyme carrier. We realized this immobilized enzyme catalyst, pectinase preparation@ZSM-5, via a simple combined strategy involving the van der Waals adsorption of pectinase preparation followed by crosslinking of the adsorbed pectinase preparation with glutaraldehyde over ZSM-5. Conformal pectinase preparation coverage of various ZSM-5 supports was achieved for the as-prepared pectinase preparation@ZSM-5. The porous pectinase preparation@ZSM-5 catalyst exhibited ultra-efficient biocatalytic activity for hydrolyzing the β-glycosidic bonds in the model substrate 4-nitrophenyl β-D-glucopyranoside, with a broad operating temperature range, high thermal stability, and excellent reusability. The relative activity of pectinase preparation@ZSM-5 at a high temperature (70 °C) was nine times higher than that of free pectinase preparation. Using thermal inactivation kinetic analysis based on the Arrhenius law, pectinase preparation@ZSM-5 showed higher activation energy for denaturation (315 kJ mol−1) and a longer half-life (62 min−1) than free pectinase preparation. Moreover, a Michaelis–Menten enzyme kinetic analysis indicated a higher maximal reaction velocity for pectinase preparation@ZSM-5 (0.22 µmol mg−1 min−1). This enhanced reactivity was attributed to the microstructure of the immobilized pectinase preparation@ZSM-5, which offered a heterogeneous reaction system that decreased the substrate–pectinase preparation binding affinity and modulated the kinetic characteristics of the enzyme. Additionally, pectinase preparation@ZSM-5 showed the best ethanol tolerance among all the reported pectinase preparation-immobilized catalysts, and an activity 247% higher than that of free pectinase preparation at a 10% (v/v) ethanol concentration was measured. Furthermore, pectinase preparation@ZSM-5 exhibited potential for practical engineering applications, promoting the hydrolysis of β-glycosidic bonds in baicalin to convert it into baicalein. This was achieved with a 98% conversion rate, i.e., 320% higher than that of the free enzyme. |
topic |
immobilization pectinase β-glycosidic bond ZSM-5 zeolite heat resistance ethanol tolerance |
url |
https://www.frontiersin.org/articles/10.3389/fchem.2021.677868/full |
work_keys_str_mv |
AT canliu immobilizedcrosslinkedpectinasepreparationonporouszsm5zeolitesasreusablebiocatalystsforultraefficienthydrolysisofbglycosidicbonds AT limingzhang immobilizedcrosslinkedpectinasepreparationonporouszsm5zeolitesasreusablebiocatalystsforultraefficienthydrolysisofbglycosidicbonds AT litan immobilizedcrosslinkedpectinasepreparationonporouszsm5zeolitesasreusablebiocatalystsforultraefficienthydrolysisofbglycosidicbonds AT yuepingliu immobilizedcrosslinkedpectinasepreparationonporouszsm5zeolitesasreusablebiocatalystsforultraefficienthydrolysisofbglycosidicbonds AT weiqiantian immobilizedcrosslinkedpectinasepreparationonporouszsm5zeolitesasreusablebiocatalystsforultraefficienthydrolysisofbglycosidicbonds AT lanqingma immobilizedcrosslinkedpectinasepreparationonporouszsm5zeolitesasreusablebiocatalystsforultraefficienthydrolysisofbglycosidicbonds |
_version_ |
1721211574362832896 |