Hybrid Nanomaterials of Magnetic Iron Nanoparticles and Graphene Oxide as Matrices for the Immobilization of β-Glucosidase: Synthesis, Characterization, and Biocatalytic Properties

Orfanakis, Georgios; Patila, Michaela; Catzikonstantinou, Alexandra V.; Lyra, Kyriaki-Marina; Kouloumpis, Αntonios; Spyrou, Konstantinos; Katapodis, Petros; Paipetis, Alkiviadis S.; Rudolf, Petra; Gournis, Dimitrios; Stamatis, Haralambos

doi:10.3389/fmats.2018.00025

Cited by 36 publications

(23 citation statements)

References 48 publications

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“…A first indication about the structure of the iron nanoparticles was gained from the XRD pattern (Supplementary Materials Figure S1). The peaks appearing in the range of 20 • and 80 • can be assigned to the crystallographic Miller planes (220), (311), (400), (422), (511), (440) indicative of γ-Fe 2 O 3 phase [19]. Moreover, in order to confirm the size of the magnetic nanoparticles, AFM height images of the γ-Fe 2 O 3 nanoparticles deposited on a Si-wafer were collected (the micrographs are shown in Figure S2, Supplementary Materials).…”

Section: Synthesis and Characterization Of The Magnetic Nanoparticlesmentioning

confidence: 99%

“…These nanomatrices offer the great advantage of easy separation from the reaction mixture just by the application of an external magnetic field, thus overcoming recoverability and reusability issues so that the overall handling process is notably facilitated. So far, several enzymes have been successfully immobilized onto various types of magnetic nanoparticles [17][18][19] indicating favorable activity and stability.…”

Section: Introductionmentioning

confidence: 99%

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Development of a Four-Enzyme Magnetic Nanobiocatalyst for Multi-Step Cascade Reactions

et al. 2019

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We report the preparation, characterization and application of a novel magnetic four-enzyme nanobiocatalyst prepared by the simultaneous covalent co-immobilization of cellulase (CelDZ1), β-glucosidase (bgl), glucose oxidase (GOx) and horseradish peroxidase (HRP) onto the surface of amino-functionalized magnetic nanoparticles (MNPs). This nanobiocatalyst was characterized by various spectroscopic techniques. The co-immobilization process yielded maximum recovered enzymatic activity (CelDZ1: 42%, bgl: 66%, GOx: 94% and HRP: 78%) at a 10% v/v cross-linker concentration, after 2 h incubation time and at 1:1 mass ratio of MNPs to total enzyme content. The immobilization process leads to an increase of Km and a decrease of Vmax values of co-immobilized enzymes. The thermal stability studies of the co-immobilized enzymes indicated up to 2-fold increase in half-life time constants and up to 1.5-fold increase in their deactivation energies compared to the native enzymes. The enhanced thermodynamic parameters of the four-enzyme co-immobilized MNPs also suggested increment in their thermal stability. Furthermore, the co-immobilized enzymes retained a significant part of their activity (up to 50%) after 5 reaction cycles at 50 °C and remained active even after 24 d of incubation at 5 °C. The nanobiocatalyst was successfully applied in a four-step cascade reaction involving the hydrolysis of cellulose.

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Section: Synthesis and Characterization Of The Magnetic Nanoparticlesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Development of a Four-Enzyme Magnetic Nanobiocatalyst for Multi-Step Cascade Reactions

et al. 2019

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“…The insufficient stability of enzyme under processing conditions is the major barrier for enzyme industrial application, high temperature, or inappropriate pH value would cause the reversible or irreversible denaturation of enzyme, which would cause the losses of enzyme activity (Orfanakis et al, 2018;Liao et al, 2019). Thus, in order to check whether the prepared biocatalyst was suitable under catalytic conditions, the effects of temperature and pH value on free alcalase and alcalase@HMSS-NH 2 -Fe 3+ were further evaluated.…”

Section: Enzymatic Performance Evaluation Of Free and Immobilized Alcmentioning

confidence: 99%

Alcalase Microarray Base on Metal Ion Modified Hollow Mesoporous Silica Spheres as a Sustainable and Efficient Catalysis Platform for Proteolysis

Qing

Sun

et al. 2020

Front. Bioeng. Biotechnol.

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The industrial exploitation of protease is limited owing to its sensitivity to environmental factors and autolysis during biocatalytic processes. In the present study, the alcalase microarray (Bacillus licheniformis, alcalase@HMSS-NH 2-Metal) based on different metal ions modified hollow mesoporous silica spheres (HMSS-NH 2-Metal) was successfully developed via a facile approach. Among the alcalase@HMSS-NH 2-Metal (Ca 2+ , Zn 2+ , Fe 3+ , Cu 2+), the alcalase@HMSS-NH 2-Fe 3+ revealed the best immobilization efficiency and enzymatic properties. This tailor-made nanocomposite immobilized alcalase on a surface-bound network of amino-metal complex bearing protein-modifiable sites via metal-protein affinity. The coordination interaction between metal ion and alcalase advantageously changed the secondary structure of enzyme, thus significantly enhanced the bioactivities and thermostability of alcalase. The as-prepared alcalase@HMSS-NH 2-Fe 3+ exhibited excellent loading capacity (227.8 ± 23.7 mg/g) and proteolytic activity. Compared to free form, the amidase activity of alcalase microarray increased by 5.3-fold, the apparent kinetic constant V max /K m of alcalase@HMSS-NH 2-Fe 3+ (15.6 min −1) was 1.9-fold higher than that of free alcalase, and the biocatalysis efficiency increased by 2.1-fold for bovine serum albumin (BSA) digestion. Moreover, this particular immobilization strategy efficiently reduced the bioactivities losses of alcalase caused by enzyme leaking and autolysis during the catalytic process. The alcalase microarray still retained 70.7 ± 3.7% of the initial activity after 10 cycles of successive reuse. Overall, this study established a promising strategy to overcome disadvantages posed by free alcalase, which provided new expectations for the application of alcalase in sustainable and efficient proteolysis.

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“…The formation of a covalent bond between the support and the enzyme usually requires the presence of a chemical reagent that acts as a cross-linker between terminal functional groups on the surface of particles and the amino-acid side chains on the protein surface. For example, our team has recently reported the covalent immobilization of β-glucosidase on hybrid nanomaterials of graphene oxide-iron magnetic nanoparticles functionalized with APTES and/or oleylamine [32]. The immobilization was carried out using glutaraldehyde as the cross-linker, and the resulted biocatalysts exhibited up to 4.4-fold higher half-time constants than free enzyme, while they were able to be reused up to 12 reaction cycles, retaining up to 40% of their initial activity.…”

Section: Enzyme Immobilization On Magnetic Particlesmentioning

confidence: 99%

“…Cross-linked enzyme aggregates (CLEAs) are obtained by precipitation of the enzyme from aqueous media followed by the cross-linking of the physical aggregates using a bifunctional reagent, such as glutaraldehyde [32]. It is a simple and cheap procedure with high immobilization yields that leads to the production of very stable biocatalysts.…”

Section: Enzyme Immobilization On Magnetic Particlesmentioning

confidence: 99%

Magnetic Microreactors with Immobilized Enzymes—From Assemblage to Contemporary Applications

2018

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Microfluidics, as the technology for continuous flow processing in microscale, is being increasingly elaborated on in enzyme biotechnology and biocatalysis. Enzymatic microreactors are a precious tool for the investigation of catalytic properties and optimization of reaction parameters in a thriving and high-yielding way. The utilization of magnetic forces in the overall microfluidic system has reinforced enzymatic processes, paving the way for novel applications in a variety of research fields. In this review, we hold a discussion on how different magnetic particles combined with the appropriate biocatalyst under the proper system configuration may constitute a powerful microsystem and provide a highly explorable scope.

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Hybrid Nanomaterials of Magnetic Iron Nanoparticles and Graphene Oxide as Matrices for the Immobilization of β-Glucosidase: Synthesis, Characterization, and Biocatalytic Properties

Cited by 36 publications

References 48 publications

Development of a Four-Enzyme Magnetic Nanobiocatalyst for Multi-Step Cascade Reactions

Development of a Four-Enzyme Magnetic Nanobiocatalyst for Multi-Step Cascade Reactions

Alcalase Microarray Base on Metal Ion Modified Hollow Mesoporous Silica Spheres as a Sustainable and Efficient Catalysis Platform for Proteolysis

Magnetic Microreactors with Immobilized Enzymes—From Assemblage to Contemporary Applications

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