NanoArmoring of Enzymes: Rational Design of Polymer-Wrapped Enzymes

Chapter One: Encapsulating Proteins in Nanoparticles: Batch by Batch or One by One
Y. Liu and A. Cao

• Abstract
• 1 Introduction
• 2 “Batch-by-Batch” Encapsulation of Proteins in Silica NPs
• 3 “One-by-One” Encapsulation of Proteins in NPs
• 4 Summary and Outlook
• Acknowledgments

Chapter Two: Enzyme Adsorption on Nanoparticle Surface Probed by Highly Sensitive Second Harmonic Light Scattering
A. Das, A. Chakrabarti and P.K. Das

• Abstract
• 1 Introduction
• 2 Techniques for Probing Nanoparticle–Protein Interaction
• 3 Second Harmonic Light Scattering
• 4 Sample Preparation
• 5 Interaction of ADH With GNPs
• 6 Interaction of Insulin With GNPs
• 7 Stoichiometry of the Protein–GNP Conjugate
• 8 Summary and Future Outlook
• Acknowledgment

Chapter Three: Armoring Enzymes by Metal–Organic Frameworks by the Coprecipitation Method
M. Hou and J. Ge

• Abstract
• 1 Introduction
• 2 Equipment
• 3 Materials
• 4 Protocol
• 5 Safety
• 6 Step1: Preparation of Enzyme@MOF
• 7 Step2: Preparation of Buffers
• 8 Step3: Purification of the Product
• 9 Step4: Characterization of Enzyme@MOF
• 10 Step5: Measurement of Enzyme Loading in MOFS
• 11 Step6: Analysis of the Location of Enzymes in MOFS
• 12 Step7: Determination of Enzyme Activities
• 13 Conclusions

Chapter Four: Enzyme Armoring by an Organosilica Layer: Synthesis and Characterization of Hybrid Organic/Inorganic Nanobiocatalysts
M.R. Correro, S. Sykora, P.F.-X. Corvini and P. Shahgaldian

• Abstract
• 1 Introduction
• 2 Equipments and Chemicals
• 3 Safety Guidelines
• 4 Procedures
• 5 Conclusions
• Acknowledgments

Chapter Five: Strategies for Biophysical Characterization of Protein–Polymer Conjugates
C. Williams, M.L. Dougherty, K. Makaroff, J. Stapleton, D. Konkolewicz, J.A. Berberich and R.C. Page

• Abstract
• 1 Introduction
• 2 Biophysical Methods
• 3 Conclusions
• Acknowledgments

Chapter Six: Guide to the Preparation of Molecularly Imprinted Polymer Nanoparticles for Protein Recognition by Solid-Phase Synthesis
J. Xu, P.X. Medina-Rangel, K. Haupt and B.Tse Sum Bui

• Abstract
• 1 Introduction
• 2 Protocols
• 3 Physicochemical Characterization of MIP-NPs
• 4 Evaluation of the Binding Characteristics of MIP-NPs Using a Quartz Crystal Microbalance
• 5 MIP-NPs Protect Enzyme From Thermal and pH Denaturation
• 6 Concluding Remarks
• 7 Notes
• Acknowledgments

Chapter Seven: Armored Urease: Enzyme-Bioconjugated Poly(acrylamide) Hydrogel as a Storage and Sensing Platform
K.R. Kunduru, S.N.R. Kutcherlapati, D. Arunbabu and T. Jana

• Abstract
• 1 Introduction
• 2 Reversible Immobilization
• 3 Jack Bean Urease: Active Site
• 4 Immobilization of Jack Bean Urease
• 5 Methodology
• 6 Results and Discussion
• 7 Conclusions and Future Perspectives
• Acknowledgments

Chapter Eight: Armored Enzyme–Nanohybrids and Their Catalytic Function Under Challenging Conditions
O.V. Zore, R.M Kasi and C.V. Kumar

• Abstract
• 1 Introduction
• 2 Methods: Synthesis of Bienzyme–Polymer Conjugates
• 3 Methods: Synthesis of Bienzyme–Polymer–Graphene Oxide (GOx–HRP–PAA/GO) Hybrid Materials
• 4 Characterization
• 5 Conclusions
• Acknowledgments

Chapter Nine: Approaches for Conjugating Tailor-Made Polymers to Proteins
M. Paeth, J. Stapleton, M.L. Dougherty, H. Fischesser, J. Shepherd, M. McCauley, R. Falatach, R.C. Page, J.A. Berberich and D. Konkolewicz

• Abstract
• 1 Introduction
• 2 Polymer Synthetic Procedures
• 3 Protein Conjugation Methods—Amine Based
• 4 Polymer Conjugation by GT Using Click Approaches
• 5 Conjugate Characterization Methods
• 6 Conclusions

Chapter Ten: NanoArmoring of Enzymes by Polymer-Functionalized Iron Oxide Nanoparticles
G. Premaratne, L. Coats and S. Krishnan

• Abstract
• 1 Introduction
• 2 NanoArmoring by Making Polymer–Enzyme Conjugates
• 3 MNP–Polymer–Enzyme Conjugates
• 4 Covalent vs Noncovalent Immobilization of Peroxidase Active Proteins on PolyMNPs. What Difference Does It Make in the Electrocatalytic Activity and Kinetics?
• 5 Effect of MNP Size on Activity and Recovery of GOx (Park, McConnell, Boddohi, Kipper, & Johnson, 2011)
• 6 Summary
• 7 Future Outlook
• Acknowledgments

Chapter Eleven: Expression of Cellulolytic Enzyme as a Fusion Protein That Reacts Specifically With a Polymeric Scaffold
P. Katyal, Y. Yang, O. Vinogradova and Y. Lin

• Abstract
• 1 Introduction
• 2 Materials
• 3 Methods
• 4 Future Outlook
• Acknowledgments

Chapter Twelve: Nanoarmoring of Proteins by Conjugation to Block Copolymer Micelles
N. Suthiwangcharoen and R. Nagarajan

• Abstract
• 1 Introduction
• 2 Synthesis of Block Copolymer–Protein (BSA) Conjugate
• 3 Characterization of Block Copolymer–Protein Conjugate
• 4 Characterization of Protein–Polymer Conjugate Micelle
• 5 Conclusions
• Acknowledgments

Chapter Thirteen: Semisynthetic Enzymes by Protein–Peptide Site-Directed Covalent Conjugation: Methods and Applications
J.M Palomo

• Abstract
• 1 Introduction
• 2 Protein–Peptide Site-Directed Covalent Conjugation
• 3 The New Semisynthetic Lipases in Biotransformations
• 4 Conclusions
• Acknowledgments

Chapter Fourteen: Transgultaminase-Mediated Nanoarmoring of Enzymes by PEGylation
A. Grigoletto, A. Mero, K. Maso and G. Pasut

• Abstract
• 1 Introduction
• 2 Materials and Reagents
• 3 Methods

Chapter Fifteen: Polymer-Based Protein Engineering: Synthesis and Characterization of Armored, High Graft Density Polymer–Protein Conjugates
S. Carmali, H. Murata, C. Cummings, K. Matyjaszewski and A.J. Russell

• Abstract
• 1 Introduction
• 2 Protocols
• 3 Protein–Polymer-Based Engineering
• 4 Conclusions
• Acknowledgment

Chapter Sixteen: Nano-Armoring of Enzymes: Rational Design of Polymer-Wrapped Enzymes
K. Raghupathi and S. Thayumanavan

• Abstract
• 1 Introduction
• 2 Covalent Conjugation
• 3 Electrostatic Complexation
• 4 Noncovalent Entrapment
• 5 Conclusions
• Acknowledgments

Chapter Seventeen: Nanoarmored Enzymes for Organic Enzymology: Synthesis and Characterization of Poly(2-Alkyloxazoline)–Enzyme Conjugates
M. Leurs and J.C. Tiller

• Abstract
• 1 Introduction
• 2 Synthesis of Poly(2-Alkyloxazoline)s With an NH₂ End Group
• 3 Conjugation of Enzymes With Poly(2-Alkyloxazoline)s
• 4 Conjugate Characterization
• 5 Activity Evaluation of Poly(2-Alkyloxazoline)–Enzyme Conjugates
• 6 Conclusions
• Acknowledgments

Chapter Eighteen: Preparation and Applications of Dendronized Polymer–Enzyme Conjugates
A. Küchler, D. Messmer, A.D. Schlüter and P. Walde

• Abstract
• 1 Introduction
• 2 The Denpol de-PG2
• 3 The Denpol–Enzyme Conjugate de-PG2-BAH-proK
• 4 Synthesis and Characterization of Denpols Carrying Other Types of Enzymes
• 5 Immobilization of de-PG2-BAH-proK on Silicate Surfaces
• 6 Entrapment of Denpol–Enzyme Conjugates Inside Phospholipid Vesicles
• 7 Conclusions and Outlook
• Acknowledgments

Chapter Nineteen: Nanoarmoring of Enzymes by Interlocking in Cellulose Fibers With Poly(Acrylic Acid)
C.M. Riccardi, R.M. Kasi and C.V. Kumar

• Abstract
• 1 Introduction
• 2 Equipment and Reagents
• 3 Methods
• 4 Conclusions
• Acknowledgments