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Siderophore and Related Natural Products Biosynthesis
- 1st Edition, Volume 702 - August 20, 2024
- Editor: Timothy Wencewicz
- Language: English
- Hardback ISBN:9 7 8 - 0 - 4 4 3 - 2 9 6 7 8 - 9
- eBook ISBN:9 7 8 - 0 - 4 4 3 - 2 9 6 7 9 - 6
Methods in Enzymology Volume 702 highlights new advances in the field, with this new volume presenting interesting topics on Siderophore and related natural products biosynthe… Read more
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Request a sales quoteMethods in Enzymology Volume 702 highlights new advances in the field, with this new volume presenting interesting topics on Siderophore and related natural products biosynthesis
- Provides the authority and expertise of leading contributors from an international board of authors
- Presents the latest release in Methods in Enzymology
- Updated release includes the latest information on Siderophore and related natural products biosynthesis
Academic, government and industrial sectors
- Cover image
- Title page
- Table of Contents
- Series Page
- Copyright
- Contributors
- Chapter One: Kinetic analysis of the three-substrate reaction mechanism of an NRPS-independent siderophore (NIS) synthetase
- Abstract
- 1 Introduction
- 2 Protein production and purification
- 3 NADH coupled adenylation assay
- 4 Kinetic analysis to distinguish among potential mechanisms
- 5 Alternate approaches
- 6 Summary and conclusions
- Acknowledgment
- References
- Chapter Two: Experimental methods for evaluating siderophore–antibiotic conjugates
- Abstract
- 1 Introduction
- 2 Methods
- 3 Summary
- References
- Chapter Three: A continuous fluorescence assay to measure nicotianamine synthase activity
- Abstract
- 1 Introduction
- 2 Assay design
- 3 Before you begin
- 4 Key resources table
- 5 Methods and equipment
- 6 Preparation of reagents
- 7 Step-by-step method details
- 8 Quantification and statistical analysis
- 9 Summary
- References
- Chapter Four: ITC-based kinetics assay for NIS synthetases
- Abstract
- 1 Introduction
- 2 General overview of the method
- 3 Before you begin
- 4 Materials and equipment
- 5 Step-by-step method details
- 6 Expected outcomes
- 7 Quantification and statistical analysis
- 8 Optimization and troubleshooting
- 9 Summary
- References
- Chapter Five: An in vitro assay to explore condensation domain specificity from non-ribosomal peptide synthesis
- Abstract
- 1 Introduction
- 2 General method and statistical analysis
- 3 Molecular design of PCP2-C3 SpyCatcher and PCP3 SpyTag constructs
- 4 Protein expression and purification
- 5 Synthesis of chemical reagents
- 6 In vitro reconstitution of NRPS C-domain
- 7 LC-HRMS/MS analysis of methylamine cleaved peptide products
- 8 Intact protein PPant ejection LC-ESI-Q-TOF-MS analysis of chemically stabilised peptide products
- 9 Conclusions
- Acknowledgements
- References
- Chapter Six: The production of siderophore analogues using precursor-directed biosynthesis
- Abstract
- 1 Introduction
- 2 Materials and equipment
- 3 Protocol
- 4 Summary
- Acknowledgements
- References
- Chapter Seven: Preparation of coenzyme F430 biosynthetic enzymes and intermediates
- Abstract
- 1 Introduction
- 2 Expression and purification of the coenzyme F430 biosynthesis enzymes
- 3 Synthesis and purification of coenzyme F430 biosynthetic intermediates
- 4 Concluding remarks
- Acknowledgments
- References
- Chapter Eight: Purification and biochemical characterization of methanobactin biosynthetic enzymes
- Abstract
- 1 Introduction
- 2 Expression and purification of M. trichosporium OB3b MbnBC complexes
- 3 In vitro modification of MbnA using purified MbnBC
- 4 Crystallization and structure determination of M. trichosporium OB3b MbnBC
- 5 Expression and purification of M. trichosporium OB3b MbnN
- 6 Conclusions
- Acknowledgments
- References
- Chapter Nine: Discovery, isolation, and characterization of diazeniumdiolate siderophores
- Abstract
- 1 Introduction
- 2 Bioinformatics and genome mining to predict bacteria producing graminine-containing siderophores
- 3 Bacterial growth conditions for production of siderophores
- 4 Detection and isolation of the siderophores
- 5 Identification and characterization of graminine-containing siderophores
- 6 Conclusions and future outlook
- Acknowledgements
- References
- Chapter Ten: Linking biosynthetic genes to natural products using inverse stable isotopic labeling (InverSIL)
- Abstract
- 1 Introduction
- 2 Key resources
- 3 Equipment and reagents
- 4 Method
- 5 Notes
- Acknowledgments
- References
- Chapter Eleven: 4-Aldrithiol-based photometric assay for detection of methylthioalkylmalate synthase activity
- Abstract
- 1 Introduction
- 2 Using continuous enzyme assays for MAMS activity
- 3 Analysis and interpretation of spectrophotometric enzyme assay for Brassica juncea MAMS isoforms
- 4 Spectrophotometric alternatives to the 4,4′-DTP assay
- 5 Conclusion and outlook
- References
- Chapter Twelve: Methods for biochemical characterization of flavin-dependent N-monooxygenases involved in siderophore biosynthesis
- Abstract
- 1 Introduction
- 2 NADPH oxidation
- 3 Oxygen consumption assays
- 4 Iodine oxidation assay
- 5 Determination of hydrogen peroxide formation and reaction uncoupling
- 6 Fmoc-Cl derivatization
- 7 Determination of reaction stereospecificity
- 8 Fluorescence anisotropy
- 9 Targeting flavin dynamics for drug discovery
- 10 Summary and conclusions
- Acknowledgments
- References
- Chapter Thirteen: Siderophore-dependent ferrichelatases
- Abstract
- 1 Introduction
- 2 General methods and statistical analysis
- 3 Overexpression and purification of N-His6-FhuD2
- 4 FhuD2 binding assay: intrinsic tryptophan fluorescence quenching
- 5 FhuD2 binding assay: siderophore affinity chromatography
- 6 Synthesis of DFO-NBD and Fe(III)-siderophores
- 7 Ferrichelatase assay
- 8 Siderophore EDTA competition assay
- 9 S. aureus growth studies
- 10 Summary and conclusions
- Acknowledgments
- References
- Chapter Fourteen: Native metabolomics for mass spectrometry-based siderophore discovery
- Abstract
- 1 Introduction
- 2 General methods and assembly of instrumentation
- 3 Sample preparation for native metabolomics
- 4 Mass spectrometry acquisition parameters
- 5 Data analysis using ion identity molecular networking
- 6 Limitations and considerations
- 7 Summary and conclusions
- Acknowledgments
- References
- Chapter Fifteen: Anaerobic heme recycling by gut microbes: Important methods for monitoring porphyrin production
- Abstract
- 1 Introduction
- 2 Methods
- 3 Procedure
- References
- Chapter Sixteen: Predicting metallophore structure and function through genome mining
- Abstract
- 1 Introduction
- 2 Materials and methods
- 3 Metallophore genomics: from sequence to molecule to function
- 4 Predicting metallophore pathways in antiSMASH
- 5 Predicting metallophore structure through comparative analyses
- 6 Next steps for the reader
- Acknowledgments
- References
- No. of pages: 412
- Language: English
- Edition: 1
- Volume: 702
- Published: August 20, 2024
- Imprint: Academic Press
- Hardback ISBN: 9780443296789
- eBook ISBN: 9780443296796
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