Enzymes of energy technology / edited by Fraser Armstrong.

Format:

Book

Contributor:

Armstrong, Fraser, editor.

Language:

English

Subjects (All):

Enzymes--Industrial applications.

Enzymes.

Physical Description:

1 online resource (404 pages)

Place of Publication:

Cambridge, MA : Academic Press, [2018]

Summary:

Enzymes of Energy Technology, Volume 613 in the Methods in Enzymology series, highlights new advances in the field, with this updated volume presenting interesting chapters written by an international board of authors. Chapters include Purification of fully active and crystallizable photosystem II from thermophilic cyanobacteria, Production and manipulation of [NiFeSe]-hydrogenases for renewable hydrogen research, Hydrogen production by [FeFe]-hydrogenases, Production and properties of enzymes that activate and produce carbon monoxide, Recombinant [NiFe]-hydrogenases from E. coli, Working with nitrogenase, Oxygen--tolerant [NiFe]-hydrogenases, Cytoplasmic and Membrane Bound Hydrogenases from the hyperthermophile Pyrococcus furiosus, and more.Additional sections cover Enzymatic conversion of methane into useful chemicals, Production and investigations of trans-membrane electron transfer protein, Characterization of post-translational modifications in methyl-coenzyme M reductase in diverse methanogens by mass-spectrometry, Reductive activation of carbon dioxide by formate dehydrogenases, and Lytic polysaccharide monooxygenases in biofuel processing.14. Production and manipulation of blue copper oxidases for technological applicationsYasmina Mekmouche- Provides the authority and expertise of leading contributors from an international board of authors- Presents the latest release in the Methods in Enzymology series- Includes the latest information on the Enzymes of Energy Technology

Contents:

Front Cover

Enzymes of Energy Technology

Copyright

Contents

Contributors

Preface

Chapter One: Purification of fully active and crystallizable photosystem II from thermophilic cyanobacteria

1. Introduction

2. Growth and harvest of cells

3. Isolation of thylakoid membranes

4. Isolation of crude PSII particles

5. Purification of PSII dimers

6. Characterization of the purified PSII core dimers

7. Crystallization of purified PSII dimers

Acknowledgments

References

Chapter Two: Production and manipulation of blue copper oxidases for technological applications

2. Heterologous production of fungal laccases

2.1. Cloning

2.1.1. Site-directed mutagenesis

2.1.2. Yeast recombination

2.2. Transformations

2.2.1. S. cerevisiae

2.2.2. A. niger

2.3. Screening for laccase secreting transformants in S. cerevisiae and A. niger

2.3.1. Screening of clones from deep-well or microtiter plate cultures

2.3.2. Screening of laccase producing transformants on petri dish plates

2.3.3. Screening of activity on 96-well plate

2.3.4. ELISA screening

2.4. Fermentation in A. niger

2.4.1. Preparation of spore inoculum and storage

2.4.2. Production in bioreactor and growth parameters

3. General purification procedure and characterization

3.1. Purification procedure

3.1.1. Assay procedures

3.1.2. Laccase purification procedure

4. Applications

4.1. General procedure for an oriented covalent grafting of the laccase surface

4.1.1. Preparation of laccase variants for single-site anchoring

4.1.2. Reductive amination of lysine

4.1.3. Evaluation of the grafting

4.2. Hybridization with materials

5. Conclusions

Acknowledgment

Further reading.

Chapter Three: Production and spectroscopic characterization of lytic polysaccharide monooxygenases

2. Expression and purification of LPMOs

2.1. Recombinant expression and purification from the E. coli periplasm

2.2. Recombinant expression and purification from the E. coli cytoplasm

2.3. Recombinant expression and purification from yeast

2.4. Recombinant expression and purification from fungi

3. EPR spectroscopy

3.1. EPR sample preparation for LPMOs

3.2. Multifrequency collection

3.3. Simulation procedures

Further reading

Chapter Four: A novel overproduction system for the structural determination of a proton-pumping hydrogen-producing [NiFe ...

1.1. Hydrogen catalysis in nature

1.2. Escherichia coli hydrogenase-2 genetic and structural arrangement

1.3. First approaches to overexpression of a [NiFe]-hydrogenase

2. Overproduction of fully formed holo-Hyd-2

2.1. Molecular biology strategies for the overproduction of Hyd-2

2.1.1. Design of overexpression plasmid pO

2.1.1.1. Equipment

2.1.1.2. Reagents and buffers

2.1.1.3. Procedures

2.1.2. Design of overexpression strain HJ001-hyp

2.2. Purification of native Hyd-2 and overproduced Hyd-2: A comparison

2.2.1. Equipment

2.2.2. Reagents and buffers

2.2.3. Procedures

2.2.4. Notes

2.3. Extension of the Hyd-2 overproduction purification protocol for crystal structure determination

2.3.1. Equipment

2.3.2. Reagents and buffers

2.3.3. Procedures

3. Biochemical and electrochemical analysis of overproduced Hyd-2 reveals fully formed and active Hyd-2-OP

4. Structural characterization leading to a proposal for the full membrane-bound structure of Hyd-2

References.

Chapter Five: O2-tolerant [NiFe]-hydrogenases of Ralstonia eutropha H16: Physiology, molecular biology, purification, and ...

2. Physiology

2.1. Lithoautotrophic cultivation of R. eutropha derivatives

2.1.1. Equipment

2.1.2. Buffers and reagents

2.1.3. Procedure

2.1.4. Notes

2.2. Heterotrophic cultivation of R. eutropha derivatives

2.2.2. Buffers and reagents

2.2.3. Procedure

3. Molecular biology

3.1. Conjugative plasmid transfer to R. eutropha

3.1.1. Equipment

3.1.2. Buffers and reagents

3.1.3. Procedure

3.2. Transformation of R. eutropha by electroporation

3.2.1. Equipment

3.2.2. Buffers and reagents

3.2.3. Procedure

3.2.4. Notes

3.3. Generation of R. eutropha mutants by double homologous recombination

3.3.1. Equipment

3.3.2. Buffers and reagents

3.3.3. Procedure

3.3.4. Notes

3.4. Plasmids replicating in R. eutropha

4. Hydrogenase purification

4.1. Purification of the soluble NAD-reducing [NiFe]-hydrogenase (SH)

4.1.1. Equipment

4.1.2. Buffers and reagents

4.1.3. Procedure

4.1.4. Notes

4.2. Purification of membrane-bound [NiFe]-hydrogenase

4.2.1. Equipment

4.2.2. Buffers and reagents

4.2.3. Procedure for heterodimeric MBH

4.2.4. Procedure for heterotrimeric MBH

4.2.5. Notes

5. Biochemical analyses

5.1. H2-dependent reduction of NAD

5.1.1. Equipment

5.1.2. Buffers and reagents

5.1.3. Procedure

5.1.4. Notes

5.2. H2-dependent reduction of redox dyes

5.2.1. Equipment

5.2.2. Buffers and reagents

5.2.3. Procedure

5.2.4. Notes

5.3. H2 evolution measurements with modified Clark electrode

5.3.1. Equipment

5.3.2. Buffers and reagents

5.3.3. Procedure

5.3.4. Notes

5.4. H2 evolution measurements by gas chromatography

5.4.1. Equipment.

5.4.2. Buffers and reagents

5.4.3. Procedure

5.5. Catalytic hydrogen-deuterium exchange measurements

5.5.1. Equipment

5.5.2. Buffers and reagents

5.5.3. Procedure

5.5.4. Notes

6. Summary and conclusion

Chapter Six: Cytoplasmic and membrane-bound hydrogenases from Pyrococcus furiosus

2. Expression strategy

3. Large-scale growth of recombinant P. furiosus strains

4. Purification of SHI

4.1. Equipment

4.2. Buffers and reagents

4.3. Procedure

5. Purification of C-MBH and S-MBH

5.1. Equipment

5.2. Buffers and reagents

5.3. Procedure

6. Assays

6.1. Equipment

6.2. Buffers and reagents

6.3. Procedure

6.3.1. Hydrogen evolution activity

6.3.2. Hydrogen oxidation activity

6.3.3. Oxygen-stability and thermostability

7. Catalytic properties of SHI and MBH

8. Summary and conclusions

Chapter Seven: Characterization of the [NiFeSe] hydrogenase from Desulfovibrio vulgaris Hildenborough

2. Preparation of the native [NiFeSe] hydrogenase from D. vulgaris Hildenborough

2.1. Growth of D. vulgaris Hildenborough for expression of the [NiFeSe] hydrogenase

2.2. Purification of the membrane form of the [NiFeSe] hydrogenase

2.3. Purification of the soluble form of the [NiFeSe] hydrogenase

3. Production of the recombinant D. vulgaris Hildenborough [NiFeSe] hydrogenase

3.1. Creation of the deletion mutant for the [NiFeSe] hydrogenase

3.2. Expression vector and complemented strain for [NiFeSe] hydrogenase expression

3.3. Site-directed mutagenesis of the [NiFeSe] hydrogenase

3.4. Purification of the recombinant [NiFeSe] hydrogenase

4. Hydrogenase activities

4.1. H2 evolution and uptake with artificial mediators

4.1.1. H2 evolution assay

4.1.2. H2 uptake assay.

4.1.3. Activity-stained native gels

4.2. H/D isotope exchange activity

4.3. Electrocatalytic H2 production and oxidation

4.4. Photocatalytic H2 production

5. Structure determination of D. vulgaris Hildenborough [NiFeSe] hydrogenase by X-ray crystallography

5.1. Crystallization

5.2. Data collection and structure determination

5.3. Structure refinement

6. D. vulgaris Hildenborough [NiFeSe] hydrogenase biophysical characterization

6.1. Infrared spectroscopy

6.2. Atomic force microscopy

6.3. Quartz crystal microbalance

Chapter Eight: [FeFe]-hydrogenases from green algae

2. Discovery and isolation of novel algal [FeFe]-hydrogenases

2.1. Step 1: Screening of H2-producing microalgae

2.1.1. Anaerobic induction

2.1.2. Sulfur deprivation

2.2. Step 2: Heterologous expression in bacterial systems

2.2.1. Promising alterations of expression protocol

Protein toxicity

Expression strain/codon usage

New media formulation

Lower expression temperatures

Tuneable expression

2.3. Step 3: Purification

Avoid any contamination with O2

Be sure to lyse cells properly

Work fast and keep protein cold

Check if the enzyme is light sensitive

Screen for appropriate storage conditions

2.4. Step 4: H-cluster assembly

2.4.1. In vitro maturation

3. Methods of characterization

3.1. IR spectroscopy

3.2. In vitro activity assays

3.2.1. In vitro activity assay H2 production

4. Conclusion and outlook

Chapter Nine: Application of affinity purification methods for analysis of the nitrogenase system from Azotobacter vinelandii

2. The nitrogenase systems in A. vinelandii

3. Strategies for application of affinity purification to the nitrogenase system

4. Methods.

4.1. Growth and cell-free extract preparation of A. vinelandii.

Notes:

Description based on print version record.

ISBN:

0-12-816362-3

0-12-816361-5