Rational design of enzyme-nanomaterials / edited by Chalia Vijaya Kumar.
Rational Design of Enzyme-Nanomaterials, the new volume in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers research methods in rational design of enzyme-nanomaterials, and includes sections on su...
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Full Text (via ScienceDirect) |
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| Format: | eBook |
| Language: | English |
| Published: |
Cambridge, MA :
Academic Press is an imprint of Elsevier,
2016.
|
| Series: | Methods in enzymology ;
v. 571. |
| Subjects: |
Table of Contents:
- Front Cover; Rational Design of Enzyme-Nanomaterials; Copyright; Contents; Contributors; Preface; Acknowledgments; Chapter One: Preparation of Biocatalytic Microparticles by Interfacial Self-Assembly of Enzyme-Nanoparticle Conjugates Aro ... ; 1. Theory; 2. Equipment; 3. Materials; 3.1. Buffer Preparation; 4. Protocol; 4.1. Duration; 4.2. Preparation; 5. Step 1: Nanoparticle Synthesis; 5.1. Overview; 5.2. Duration; 5.3. Tip; 5.4. Tip; 5.5. Tip; 5.6. Tip; 5.7. Tip; 6. Step 2: Purification of Enzyme; 6.1. Overview; 6.2. Duration; 6.3. Tip; 6.4. Tip.
- 7. Step 3: Preparation of the Aqueous Phase and Oil Phase7.1. Overview; 7.2. Duration; 7.3. Tip; 7.4. Tip; 7.5. Tip; 7.6. Tip; 7.7. Tip; 7.8. Tip; 7.9. Tip; 8. Step 4: Microparticle Assembly; 8.1. Overview; 8.2. Duration; 8.3. Tip; 8.4. Tip; 8.5. Tip; 9. Step 5: Microparticle Washing; 9.1. Overview; 9.2. Duration; 9.3. Tip; 9.4. Tip; 10. Conclusions; References; Chapter Two: Monitoring Enzymatic Proteolysis Using Either Enzyme- or Substrate-Bioconjugated Quantum Dots; 1. Introduction; 1.1. Enzyme-Nanoparticle Constructs.
- 1.2. Quantification Assay for Observing Modified Kinetics with Enzyme-QD Conjugates1.3. Enzyme Activity Sensors Based on Transient QD-Enzyme Interactions; 2. Quantification Assay for Observing Modified Kinetics with Enzyme-QD Conjugates; 2.1. Materials; 2.1.1. Equipment; 2.1.2. Reagents; 2.2. Enzyme Assembly onto QDs; 2.3. Obtaining Kinetic Data of QD-Enzyme Constructs; 2.4. Analyzing the Data; 3. Enzyme Activity Sensors Based on Transient QD-Enzyme Interactions; 3.1. Materials; 3.1.1. General Equipment; 3.1.2. Peptide Labeling; 3.1.2.1. Equipment; 3.1.2.2. Reagents.
- 3.1.3. Peptide Precleaving3.1.3.1. Equipment; 3.1.3.2. Reagents; 3.1.4. QD-Peptide Construct; 3.1.4.1. Equipment; 3.1.4.2. Reagents; 3.1.5. Spectral Characterization and Kinetics Measurements; 3.1.5.1. Equipment; 3.1.5.2. Reagents; 3.2. Peptide Labeling; 3.2.1. Single Cysteine Labeling; 3.2.2. Dual Labeling for Control Experiments; 3.2.3. Peptide Purification, Quantification, and Storage; 3.3. Peptide Precleaving; 3.4. QD-Peptide Construct; 3.4.1. Formation; 3.4.2. Characterization; 3.5. Spectral Characterization; 3.6. Fixed Enzyme Experiments; 3.7. Fixed Substrate Experiments.
- 3.8. Data Analysis4. Notes; Acknowledgments; References; Chapter Three: Intense PEGylation of Enzyme Surfaces: Relevant Stabilizing Effects; 1. Introduction; 2. Theory; 3. Protocols; 3.1. PEGylation of Chemically Aminated Enzymes; 3.1.1. Rhizomucor miehei Lipase; 3.1.2. Thermomyces lanuginosa Lipase; 3.2. PEGylation of Enzymes Coated with Polymers; 3.2.1. Candida antarctica B Lipase; 3.2.2. Bioxilanase L Plus (Trichoderma reesei endo-1,4-β-Xylanase); 4. Inactivation of Modified Enzyme Derivatives; 5. Conclusions; Acknowledgments; References.