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Applied Biochemistry and Biotechnology
Article . 2012 . Peer-reviewed
License: Springer TDM
Data sources: Crossref
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Improved Covalent Immobilization of Horseradish Peroxidase on Macroporous Glycidyl Methacrylate-Based Copolymers

Authors: Olivera, Prodanović; Miloš, Prokopijević; Dragica, Spasojević; Zeljko, Stojanović; Ksenija, Radotić; Zorica D, Knežević-Jugović; Radivoje, Prodanović;

Improved Covalent Immobilization of Horseradish Peroxidase on Macroporous Glycidyl Methacrylate-Based Copolymers

Abstract

A macroporous copolymer of glycidyl methacrylate and ethylene glycol dimethacrylate, poly(GMA-co-EGDMA), with various surface characteristics and mean pore size diameters ranging from 44 to 200 nm was synthesized, modified with 1,2-diaminoethane, and tested as a carrier for immobilization of horseradish peroxidase (HRP) by two covalent methods, glutaraldehyde and periodate. The highest specific activity of around 35 U g(-1) dry weight of carrier was achieved on poly(GMA-co-EGDMA) copolymers with mean pore diameters of 200 and 120 nm by the periodate method. A study of deactivation kinetics at 65 °C and in 80 % dioxane revealed that periodate immobilization also produced an appreciable stabilization of the biocatalyst, while stabilization factor depended strongly on the surface characteristics of the copolymers. HRP immobilized on copolymer with a mean pore diameter of 120 nm by periodate method showing not only the highest specific activity but also good stability was further characterized. It appeared that the immobilization resulted in the stabilization of enzyme over a broader pH range while the Michaelis constant value (K (m)) of the immobilized HRP was 10.8 mM, approximately 5.6 times higher than that of the free enzyme. After 6 cycles of repeated use in a batch reactor for pyrogallol oxidation, the immobilized HRP retained 45 % of its original activity.

Country
Serbia
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Subjects by Vocabulary

Microsoft Academic Graph classification: Glycidyl methacrylate Methacrylate Horseradish peroxidase chemistry.chemical_compound Polymer chemistry Copolymer biology Chemistry Periodate Covalent bond biology.protein Glutaraldehyde Ethylene glycol

Keywords

Morphology, Polymers, Surface Properties, Bioengineering, Pyrogallol, Applied Microbiology and Biotechnology, Biochemistry, Horseradish peroxidase, Covalent immobilization, Copolymerization, Biological applications of polymers, Molecular Biology, Periodic Acid, Hydrogen Bonding, General Medicine, Enzymes, Immobilized, Enzymes, Kinetics, Glutaral, Macroporous polymers, Epoxy Compounds, Methacrylates, Ethylene Glycols, Biotechnology

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download
citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
views
OpenAIRE UsageCountsViews provided by UsageCounts
downloads
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21
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152
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