Methods in Methane Metabolism Part B Methanotrophy 1st Edition by Amy Rosenzweig, Stephen W Ragsdale ISBN 0123869056 9780123869050

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Product details: 

ISBN 10: 0123869056  

ISBN 13: 9780123869050

Author: Amy Rosenzweig, Stephen W. Ragsdale

Produced by microbes on a large scale, methane is an important alternative fuel as well as a potent greenhouse gas. This volume focuses on microbial methane metabolism, which is central to the global carbon cycle. Both methanotrophy and methanogenesis are covered in detail. Topics include isolation and classification of microorganisms, metagenomics approaches, biochemistry of key metabolic enzymes, gene regulation and genetic systems, and field measurements. The state of the art techniques described here will both guide researchers in specific pursuits and educate the wider scientific community about this exciting and rapidly developing field.

Topics include isolation and classification of microorganisms, metagenomics approaches, biochemistry of key metabolic enzymes, gene regulation and genetic systems and field measurements.

The state-of-the-art techniques described here will both guide researchers in specific pursuits and educate the wider scientific community about this exciting and rapidly developing field.

Table of contents: 

Chapter 1: Characterization of the Recombinant Pyrophosphate-Dependent 6-Phosphofructokinases from M
1. Introduction
2. Methods
3. Brief Overview of Current Knowledge of PPi-PFK in Microorganisms
4. Properties of PPi-PFK from Methanotrophs
5. Conclusion
Acknowledgments
References
Chapter 2: Genes and Enzymes of Ectoine Biosynthesis in Halotolerant Methanotrophs
1. Introduction
2. PCR-Based Approach for Identification of the Ectoine Biosynthesis Genes in Methanotrophs
3. Transcriptional Regulation of the Ectoine Biosynthesis Genes
4. Key Enzymes of Ectoine Biosynthesis
5. Conclusion
Acknowledgments
References
Chapter 3: Facultative and Obligate Methanotrophs: How to Identifyand Differentiate Them
1. Introduction
2. Identification of Methanotrophic Capabilities in Novel Isolates
3. Substrate Utilization Tests
4. Tests for Culture Purity
References
Chapter 4: Approaches for the Characterization and Description of Novel Methanotrophic Bacteria
1. Introduction to Aerobic Methanotroph Characterization for Systematic Research and Other Purposes
2. Broad Level Systematic Diversity
3. Methodological Approaches
4. Concluding Remarks
References
Chapter 5: Methylococcus capsulatus (Bath): From Genome to Protein Function, and Vice Versa
1. Introduction
2. Mapping of the Outer Membrane Proteome of M. capsulatus
3. Inner Membrane Protein Complexes Studied with Blue-Native Polyacrylamide Gel Electrophoresis (BN-
4. Visualization and Identification of M. capsulatus c-type Heme Proteins
Acknowledgments
References
Chapter 6: Functional Metagenomics of Methylotrophs
1. Introduction
2. Enrichment for Specific Functional Types Using SIP
3. Creating a Metagenomic Database and Linking It to Functionality
4. Ultrashort Read-Based Metatranscriptomics
5. Conclusions and Future Perspectives
Acknowledgments
References
Chapter 7: Genetic Systems for Moderately Halo(alkali)philic Bacteria of the Genus Methylomicrobium
1. Introduction
2. Methods
3. Outlook
Acknowledgments
References
Chapter 8: Development of a System for Genetic Manipulation of the Facultative Methanotroph Methyloc
1. Introduction
2. Growth of M. silvestris
3. Introduction of Plasmid DNA by Conjugation and Electroporation
4. Gene Deletion by Electroporation of Linear DNA
5. Preparation of Competent Cells for Electroporation of M. silvestris
6. Construction of Linear DNA for Gene Deletion by Homologous Recombination
7. Electroporation
8. Efficiency of Gene Deletion
9. Case Study: Deletion of Isocitrate Lyase
10. Complementation
11. Phenotype of the Isocitrate Lyase Deletion Mutant
12. Conclusions
References
Chapter 9: Mutagenesis of Soluble Methane Monooxygenase
1. Introduction
2. Bacteriological Growth Media and Antibiotics
3. Mutagenesis and Subcloning
4. Expression Hosts and Conjugation
5. Analysis of Mutants
Acknowledgments
References
Chapter 10: Single Cell Methods for Methane Oxidation Analysis
1. Introduction
2. Methods
Acknowledgment
References
Chapter 11: Methanotroph Outer Membrane Preparation
1. Introduction
2. Isolation of OMs
3. Enrichment of Integral and Tightly Associated OMPs
4. Extraction of Surface-Associated Proteins
5. Biotin Labeling of Surface-Exposed OMPs
References
Chapter 12: Overexpression and Purification of the Particulate Methane Monooxygenase from Methylococ
1. Introduction
2. Overproduction of pMMO
3. Isolation and Purification of pMMO from the pMMO-Enriched Membranes
4. Characterization of the Purified pMMO-Detergent Complex
Acknowledgments
References
Chapter 13: Metal Reconstitution of Particulate Methane Monooxygenase and Heterologous Expression of
1. Introduction
2. Preparation of apo pMMO Membranes and Metal Reconstitution
3. Soluble Domain Constructs of the pmoB Subunit
Acknowledgments
References
Chapter 14: Particulate Methane Monooxygenase from Methylosinus trichosporium OB3b
1. Introduction
2. Bacterial Cells Expressing pMMO
3. Membrane-Bound Form of pMMO
4. Purification of Detergent-Solubilized pMMO
5. Summary
Acknowledgments
References
Chapter 15: Production, Isolation, Purification, and Functional Characterization of Methanobactins
1. Introduction: Copper, Siderophores, Chalkophores, and Methanobactin
2. Methanotroph Growth and Optimizing Methanobactin Production
3. Methanobactin Isolation and Purification
4. Studying Methanobactins in Pseudonatural Environments
Acknowledgments
References
Chapter 16: A Simple Assay for Screening Microorganisms for Chalkophore Production
1. Introduction
2. Cu-CAS Assay for Chalkophore Detection
3. Fe-CAS Assay for Detecting Nonspecific Binding of Copper from Cu-CAS by a Siderophore
4. Conclusions
Acknowledgments
References
Chapter 17: Isolation of Methanobactin from the Spent Media of Methane-Oxidizing Bacteria
1. Introduction
2. Isolation of Methanobactin from the Spent Media of MOB
3. Purification of mb
4. Sample Variability
Acknowledgments
References
Chapter 18: Measurements of Biosphere-Atmosphere Exchange of CH4 in Terrestrial Ecosystems
1. Introduction
2. Chamber Measurements of CH4 Exchange
3. Micrometeorological Measurements of CH4 Exchange

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