Mononuclear Non-heme Iron Dependent Enzymes Part B
- 1st Edition, Volume 704 - September 17, 2024
- Imprint: Academic Press
- Editor: Jennifer Bridwell-Rabb
- Language: English
- Hardback ISBN:9 7 8 - 0 - 4 4 3 - 3 4 6 4 7 - 7
- eBook ISBN:9 7 8 - 0 - 4 4 3 - 3 4 6 4 8 - 4
Mononuclear Non-heme Iron Dependent Enzymes, Volume 704, Part B focuses on methods for studying, characterizing, and leveraging the chemistry of mononuclear non-heme iron depend… Read more
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Request a sales quoteMononuclear Non-heme Iron Dependent Enzymes, Volume 704, Part B focuses on methods for studying, characterizing, and leveraging the chemistry of mononuclear non-heme iron dependent enzymes. Chapters in this new release include Observing extradiol dioxygenases in action through a crystalline lens, Methods used to determine the structure of the oxygenase component of naphthalene 1,2-dioxygenase, Expression, purification, kinetics, and crystallization of non-heme mononuclear iron enzymes: biphenyl, phthalate, terephthalate dioxygenase, Substitution of the Mononuclear, Non-heme Iron Cofactor in Lipoxygenases for Structural Studies: Non-heme iron enzyme dependent reactions, and much more.
Other sections cover Cyclopropanation and aziridination catalyzed by non-heme iron and 2-oxogluatarate dependent enzymes, Expression, purification, and characterization of non-heme Iron-dependent mono-oxygenase OzmD in Oxazinomycin Biosynthesis, Isonitrile Biosynthesis by a Non-Heme Iron(II)-Dependent Oxidases/Decarboxylase, Functional analysis of an a-ketoglutarate-dependent non-heme iron oxygenase in fungal meroterpenoid biosynthesis, Spectroscopic and Computational Studies of a Bifunctional Iron- and 2-oxoglutarate Dependent Enzyme, AsqJ, and much more.
Other sections cover Cyclopropanation and aziridination catalyzed by non-heme iron and 2-oxogluatarate dependent enzymes, Expression, purification, and characterization of non-heme Iron-dependent mono-oxygenase OzmD in Oxazinomycin Biosynthesis, Isonitrile Biosynthesis by a Non-Heme Iron(II)-Dependent Oxidases/Decarboxylase, Functional analysis of an a-ketoglutarate-dependent non-heme iron oxygenase in fungal meroterpenoid biosynthesis, Spectroscopic and Computational Studies of a Bifunctional Iron- and 2-oxoglutarate Dependent Enzyme, AsqJ, and much more.
- Provides detailed articles regarding how to study the structures and mechanisms of mononuclear non-heme iron dependent enzymes
- Guides readers on how to use partner proteins in non-heme iron enzyme catalysis
- Includes strategies to employ mononuclear non-heme iron enzymes in biocatalytic applications
Chemists who study the remarkable transformations that are facilitated by metalloproteins. Scientists who are interested in purifying, crystallizing, mechanistically studying the chemistry of mononuclear non-heme iron enzymes, and/or leveraging the chemistry of these enzymes for biocatalytic endeavors
- Cover image
- Title page
- Table of Contents
- Series Page
- Copyright
- Contributors
- Section 1 Structural studies of mononuclear non-hemeiron dependent enzymes
- Chapter One: Observing extradiol dioxygenases in action through a crystalline lens
- Abstract
- 1 Introduction
- 2 Protein expression and purification
- 3 Metal reconstitution and iron content analysis
- 4 Activity assays
- 5 Crystallization and in crystallo reaction
- 6 Single-crystal and solution spectroscopies
- 7 Concluding remarks
- Acknowledgments
- References
- Chapter Two: Methods used to determine the structure of the oxygenase component of naphthalene 1,2 dioxygenase
- Abstract
- 1 Introduction
- 2 Storage of protein for crystallization and further studies
- 3 Protein crystallization
- 4 Protein crystals for initial structure determination
- 5 An occupied active site
- 6 NDO-O grown in minimal salts media
- 7 Preparation of NDO-O ligand complex crystals
- 8 Microspectrophotometry and preparation of active enzyme-substrate complexes
- 9 Conclusion
- References
- Chapter Three: Expression, purification, kinetics, and crystallization of non-heme mononuclear iron enzymes: Biphenyl, Phthalate, and Terephthalate dioxygenases
- Abstract
- 1 Introduction
- 2 Overexpression of phthalate, terephthalate, and biphenyl dioxygenases
- 3 Purification of phthalate, terephthalate, and biphenyl dioxygenases
- 4 Biochemical activity
- 5 Crystallization of phthalate, terephthalate, and biphenyl dioxygenases
- 6 Harvesting of the crystals and X-ray diffraction
- 7 Summary
- Acknowledgments
- References
- Chapter Four: Substitution of the mononuclear, non-heme iron cofactor in lipoxygenases for structural studies
- Abstract
- 1 Introduction
- 2 Applications
- 3 Procedures for metal substitution of iron for manganese in LOXs
- 4 Sample preparation for structural studies
- 5 Summary and conclusions
- References
- Section 2 Non-heme iron enzyme dependent reactions
- Chapter Five: Cyclopropanation and aziridination catalyzed by non-heme iron and 2-oxoglutarate dependent enzymes
- Abstract
- 1 Introduction
- 2 Cyclopropanation
- 3 Aziridination
- 4 Conclusions
- Acknowledgments
- References
- Chapter Six: Expression, purification and characterization of non-heme iron-dependent mono-oxygenase OzmD in oxazinomycin biosynthesis
- Abstract
- 1 Introduction
- 2 Cloning, overexpression and purification of OzmD
- 3 Characterization of OzmD protein
- 4 Activity assays
- 5 Perspective
- Acknowledgments
- References
- Chapter Seven: Isonitrile biosynthesis by non-heme iron(II)-dependent oxidases/decarboxylases
- Abstract
- 1 Introduction
- 2 Enzyme expression, purification, and in vitro assays with ScoE
- 3 Mechanistic dissection of isonitrile formation using spectroscopic methods
- 4 Enzyme expression, purification, and in vitro assays with Rv0097
- 5 Perspectives
- Acknowledgments
- References
- Chapter Eight: Functional analysis of an α-ketoglutarate-dependent non-heme iron oxygenase in fungal meroterpenoid biosynthesis
- Abstract
- 1 Introduction
- 2 Preparation of substrate
- 3 In vitro enzyme reaction
- 4 Structural analysis and mutagenesis studies
- 5 Conclusions
- Acknowledgments
- References
- Chapter Nine: Spectroscopic and computational studies of a bifunctional iron- and 2-oxoglutarate dependent enzyme, AsqJ
- Abstract
- 1 Introduction
- 2 Biochemical and spectroscopic characterization of AsqJ
- 3 X-ray protein crystallographic characterization of AsqJ
- 4 Molecular dynamics simulations
- 5 Proposed reaction mechanism
- Acknowledgments
- References
- Chapter Ten: Expression, purification, and biochemical characterization of micro- and macroalgal kainoid synthases
- Abstract
- 1 Introduction
- 2 Molecular biology of kainoid synthase constructs
- 3 Heterologous protein expression and purification of kainoid synthases
- 4 Kainoid synthase chemical assays
- 5 Conclusions and future outlook
- Acknowledgments
- References
- Chapter Eleven: Approaches to determination of the mechanism of the Rieske monooxygenase salicylate 5-hydroxylase
- Abstract
- 1 Introduction
- 2 Preparation of salicylate 5-hydroxylase
- 3 Methods and procedures
- 4 S5HH spectra
- 5 Kinetic characterization by single-turnover
- 6 Substrate range and hydroxylation site
- 7 Kinetic isotope effects
- 8 Insight into the mechanism of monooxygenation by S5H
- 9 Conclusions for the Rieske oxygenase class
- Acknowledgments
- References
- Chapter Twelve: In vitro assay and inhibition of 9-cis-epoxycarotenoid dioxygenase (NCED) from Solanum lycopersicum and Zea mays
- Abstract
- 1 Introduction
- 2 9-cis-neoxanthin isolation from spinach and characterisation by ultraviolet–visible spectroscopy (UV-Vis) and high-performance liquid chromatography (HPLC)
- 3 Overexpression of S. lycopersicum and Z. mays NCED in Escherichia coli
- 4 NCED assay and enzyme inhibition assays
- 5 Discussion
- 6 Conclusions
- Acknowledgements
- References
- Chapter Thirteen: Methods for production and assaying catalysis of isolated recombinant human aspartate/asparagine-β-hydroxylase
- Abstract
- 1 Introduction
- 2 Preparation of purified isolated human AspH
- 3 Mass spectrometry-based assays for isolated recombinant human AspH315-758
- 4 Conclusions
- References
- Chapter Fourteen: Biochemical and biophysical approaches to characterization of the aromatic amino acid hydroxylases
- Abstract
- Abbreviations
- 1 Introduction
- 2 Structure
- 3 Catalytic mechanism
- 4 Substrate specificity
- 5 Regulation
- 6 Experimental approaches
- 7 Conclusion
- References
- Edition: 1
- Volume: 704
- Published: September 17, 2024
- Imprint: Academic Press
- No. of pages: 380
- Language: English
- Hardback ISBN: 9780443346477
- eBook ISBN: 9780443346484
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