Enzyme nanoarchitectures : enzymes armored with graphene / edited by Challa V. Kumar.

Format:

Book

Contributor:

Kumar, Challa V., editor.

Series:

Methods in enzymology ; 0076-6879 Volume 609.

Methods in enzymology ; Volume 609

Language:

English

Subjects (All):

Enzymes--Biotechnology.

Enzymes.

Nanobiotechnology.

Graphene.

Physical Description:

1 online resource (422 pages).

Edition:

First edition.

Place of Publication:

Cambridge, MA : Academic Press is an imprint of Elsevier, [2018]

Summary:

Enzymes Conjugated to Graphene, Volume 609 in the Methods in Enzymology series, highlights new advances in the field, with this new volume presenting interesting chapters on Enzyme immobilization, Detection of Urea, Enzyme immobilization Enzyme immobilization, PAMAM dendrimer modified reduced graphene oxide post functionalized by horseradish peroxidase for biosensing H2O2, HRP immobilized for LEV detection, Enzyme immobilization, Graphene biocatalysts, Enzyme immobilization, Interactions, Enzyme immobilization, GQD, Enzyme Immobilization, and Enzyme immobilization on functionalized graphene oxide nanosheets.- Provides the authority and expertise of leading contributors from an international board of authors- Presents the latest release in the Methods of Enzymology series- Updated release includes the latest information on the enzymes conjugated to graphene

Contents:

Front Cover

Enzyme Nanoarchitectures: Enzymes Armored with Graphene

Copyright

Contents

Contributors

Preface

Acknowledgments

Reference

Chapter One: Interlocking Enzymes in Graphene-Coated Cellulose Paper for Increased Enzymatic Efficiency

1. Introduction

1.1. Why Use Cellulose?

1.2. Why Choose Interlocking?

1.3. Why Use Graphene?

1.4. Why Use BSA?

1.5. Why Use GOx and HRP?

2. Materials and Methods

2.1. General Equipment

2.2. Synthesis of Biographene

2.2.1. Characterization of Biographene With Raman Spectroscopy

2.3. Method for Enzyme Interlocking in Paper

2.4. Enzyme Loading on Paper by Bradford Assay

2.5. Colorimetric Enzymatic Activity Assays

2.5.1. GOx/HRP Activity Assay

2.6. Quantification of Km and Vmax

2.7. Optimization of GOx/HRP/Biographene Interlocking in Paper

2.7.1. Effect of Increased EDC Concentrations for Interlocking

2.7.2. Effect of Wash Time

3. Concluding Remarks

References

Chapter Two: Enzyme Multilayers on Graphene-Based FETs for Biosensing Applications

2. rGO-Based FETs

2.1. Fabrication

2.2. Characterization

3. Enzyme Immobilization

3.1. LbL Assembly

3.2. SPR Characterization

4. Enzymatic Biosensors

4.1. Single-Enzyme Sensor

4.2. Enzymatic Heavy Metal Sensor

5. Summary and Conclusions

Chapter Three: Stabilization of Laccase Through Immobilization on Functionalized GO-Derivatives

1.1. Laccase as a Natural Biocatalyst

1.2. Biocatalytic Systems in Nanobiotechnology

1.3. Graphene-Based Nanomaterials and Their Application in Nanobiocatalysis

2. Synthesis of Nanomaterials

2.1. Synthesis of GO

2.2. Modification of GO With Diamines and/or Amino Acids

3. Immobilization of Laccase on GO-Derivatives.

3.1. Noncovalent Immobilization of TvL on fGO

3.2. Covalent Immobilization of TvL on fGO With Terminal Amine Groups

3.3. Covalent Immobilization of TvL on fGO With Terminal Carboxyl Groups

3.4. Synthesis of Multilayer Assemblies of TvL and fGO

4. Characterization of the Immobilized Biocatalysts

4.1. Determination of Immobilization Yield

4.1.1. BCA Assay

4.1.2. Bradford Assay

4.2. Determination of Free and Immobilized Laccase Activity

4.3. Characterization of Nanobiocatalysts by FTIR Spectroscopy

4.4. Characterization of Laccase-Based Multilayer Nanoassemblies by AFM

5. Stability and Reusability of Nanobiocatalysts

5.1. Determination of the Stability of Immobilized TvL

5.2. Determination of the Reusability of Immobilized TvL

6. Conclusion

Chapter Four: Synthesis, Characterization, and Applications of Nanographene-Armored Enzymes

1.1. Synthesis of Graphene-Armored Materials

1.2. Functionalization of Graphene-Armored Materials

1.3. Advantages of Using Graphene-Based Materials in Health-Related Applications

2. Facile Synthesis of Enzyme-Graphene-Armored Nanocomposites as Biocatalysts

3. Immobilization of the Enzyme Onto Graphene Anchored Nanocomposites

3.1. Graphene-Iron Oxide NC

3.1.1. Synthesis of GrFe3O4 NC

3.1.2. Characterization of GrFe3O4 NC

3.1.3. Immobilization of β-Galactosidase on GrFe3O4 NC

3.1.4. Stability Studies of Immobilized β-Galactosidase

3.1.5. Genotoxicity Assessment of GrFe3O4 NCs

3.2. Synthesis and Characterization of GrFe3O4 NC

3.3. Binding Studies of Immobilized β-Galactosidase

3.4. Stability Studies of Immobilized β-Galactosidase

3.4.1. Effect of pH and Temperature on the Activity of Free and Immobilized β-Galactosidase

3.4.2. Effect of Galactose.

3.5. Reusability and Storage Stability

3.6. Toxicity Assessment

3.7. Polyaniline-Coated Silver-Functionalized Graphene NC

3.7.1. Synthesis of PANI/Ag/GONC

3.7.2. Characterization of PANI/Ag/GONC

3.7.2.1. Fourier-Transform Infrared Spectroscopy

3.7.2.2. Transmission Electron Microscopy

3.7.2.3. Scanning Electron Microscopy and Energy Dispersive X-Ray Spectroscopy (EDS)

3.7.2.4. AFM

3.7.2.5. DLS Measurements

3.7.3. Immobilization of Lipase on PANI/Ag/GONC (Zhu &amp

Sun, 2012)

3.7.4. Activity and Stability Studies of ANLPANI/Ag/GONC

3.7.5. Reusability Assay

3.8. Characterization of Free and ANL Bound PANI/Ag/GONC

3.9. Immobilization Efficiency of ANLPANI/Ag/GONC

3.10. Stability Studies of ANLPANI/Ag/GONC

3.11. Reusability Analysis of ANLPANI/Ag/GONC

4. Summary and Future Outlook

Funding

Further Reading

Chapter Five: PAMAM Dendrimer Modified Reduced Graphene Oxide Postfunctionalized by Horseradish Peroxidase for Biosensing ...

2. Functionalization of Graphene (or) RGO by Noncovalent Interactions

3. Covalent Functionalization of Graphene, RGO

4. Graphene in Biosensing

5. Synthesis of Carrot Extract Reduced Graphene Oxide

5.1. Equipment

5.2. Materials

5.3. Preparation Method

5.3.1. Preparation of Graphene Oxide by Hummers Method

5.3.2. Preparation of Carrot Extract

5.3.3. Synthesis of Reduced Graphene Oxide (Ct-RGO) by a Green Approach

5.4. Characterization of the Reduced Graphene Oxide Prepared by the Green Approach

5.4.1. Equipment

5.4.2. Materials

5.4.3. Procedure

5.4.4. Characterization

6. Functionalization of Graphene by PAMAM Dendrimers via Electrochemical Grafting for Enzymatic Sensing Application

6.1. Equipment

6.2. Materials

6.3. Methods

6.3.1. Electrode Preparation.

6.3.2. Modification of GCE by Ct-RGO

6.3.3. Electrochemical Grafting of PAMAM on Ct-RGO

6.3.4. Following the Process of Electrochemical Grafting by Cyclic Voltammetry

7. Postfunctionalization of GCE/Ct-RGO-PAMAM by the Horseradish Peroxidase Enzyme

7.1. Equipment

7.2. Materials

7.3. Phosphate Buffer Preparation (PBS, pH 7)

7.4. Enzyme Immobilization

8. Application of HRP-Modified GCE/Ct-RGO-PAMAM-GA-HRP for H2O2 Sensing

8.1. Equipment

8.2. Materials

8.3. Method

9. Application of HRP-Modified GCE/Ct-RGO-PAMAM-GA-HRP for H2O2 Sensing in Serum

9.1. Equipment

9.2. Materials

9.3. Method

10. Conclusions

Chapter Six: Preparation, Characterization, and Application of Enzyme Nanoparticles

1.1. Introduction to Enzyme Kinetics

1.2. What Are Nanoparticles?

1.3. Enzyme Nanoparticles

2. Experimental

2.1. Chemicals and Reagents

2.2. Equipment

2.3. Methods to Prepare ENPs

2.4. Preparation of ENPs by Desolvation

2.4.1. Desolvation

2.4.2. Cross-linking

2.4.3. Functionalization

2.4.4. Purification

2.5. Characterization of ENPs

2.5.1. Transmission Electron Microscopy

2.5.2. Colorimetric Methods

2.5.3. UV Absorption Spectra

2.5.4. FTIR Spectra

3. Immobilization of ENPs

3.1. Characterization of ENPs Immobilized Onto Matrix

3.2. Kinetic Properties of ENPs

3.2.1. Optimum pH

3.2.2. Optimum Temperature and Thermostability

3.2.3. Response Time

3.2.4. Km Values

3.2.5. Working Range

4. Stability and Reusability

4.1. Storage Stability

4.2. Reusability

4.3. Correlation of Measurements With a Standard Method

5. Applications of ENPs

5.1. Biosensor Based on Immobilization of Nanoparticles of Cholesterol Esterase and Cholesterol Oxidase.

5.2. Biosensor Based on Immobilization of Nanoparticles of Lipase, Glycerol Kinase, and Glycerol-3-Phosphate Oxidase

5.3. Biosensor Based on Immobilization of Nanoparticles of Hb

5.4. Biosensor Based on Immobilized HbNPs for the Determination of Acrylamide in Processed Foods

5.5. Biosensor Based on Immobilized HRP Nanoparticles

5.6. Biosensor Based on Immobilized GOD Nanoparticles

5.7. Biosensor Based on Immobilized ChOx Nanoparticles

5.8. Biosensor Based on Immobilized Uricase Nanoparticles

6. Detailed Procedure for the Development of HbNPs-Based Biosensors

6.1. Reagents

6.2. Instruments Used

6.3. Preparation and Testing of HbNP Biosensor

7. Conclusions and Future Perspective

Conflict of Interest

Chapter Seven: Encapsulation of Microorganisms, Enzymes, and Redox Mediators in Graphene Oxide and Reduced Graphene Oxide

2. Methods

2.2. Encapsulation of Yeast and Bacteria in GO Hydrogel

2.3. Encapsulation of Enzymes in GO

2.4. Preparation of Mediator/Nanoparticle-Modified GO Hydrogel

2.5. Electrode Modification With GO Hydrogel

2.5.1. Modification of Glassy Carbon Electrodes With GO and GO Composites

2.5.2. Filtration-Evaporation Method for Carbon Cloth-GO Electrode Fabrication

2.6. Reduction of GO to rGO

2.6.1. Electrochemical Reduction

2.6.2. Reduction of GO to rGO by Biocatalysis

2.6.3. GO Reduction by Encapsulated GOx or GOx-Displaying Yeast

2.6.4. GO Reduction by Encapsulated S. oneidensis

2.7. Electrochemical Characterization of Modified Electrodes

Chapter Eight: Chemical and Biochemical Approach to Make a Perfect Biocatalytic System on Carbonaceous Matrices

1.1. Methods of Immobilization.

1.2. Examples of the Most Promising Immobilization Matrices.

Notes:

Includes bibliographical references and index.

Description based on print version record.

ISBN:

0-12-815241-9

0-12-815240-0