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This volume provides new technologies on NRPSs and related carrier protein dependent synthases, including polyketide synthases (PKS) and fatty acid synthases (FAS). Chapters detail enzymology, structural biology, proteopromics, chemical biology, natural product chemistry, and bioinformatics. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and methods, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.
Authoritative and cutting-edge, Non-Ribosomal Peptide Biosynthesis and Engineering: Methods and Protocols aims to feature methods that will be beneficial to new researchers, and those wanting to adopt new methodologies into their research.
List of contents
The Assembly-line Enzymology of Nonribosomal Peptide Biosynthesis.- Structural Studies of Modular Nonribosomal Peptide Synthetases.- Using NMR Titration to Study Protein-Protein Interactions in AcpP Dependent FAS-II.- Chemoproteomic Profiling of Adenylation Domain Functions in Gramicidin S-producing Nonribosomal Peptide Synthetases.- In Vitro Biochemical Characterization of Excised Macrocyclizing Thioesterase Domains from Non-Ribosomal Peptide Synthetases.- Chemo-Enzymatic Synthesis of Non-Ribosomal Macrolactams by a Penicillin-binding Protein-type Thioesterase.- Directed Evolution of the BpsA Carrier Protein Domain for Recognition by Non-cognate 4'-Phosphopantetheinyl Transferases to Enable Inhibitor Screening.- Unraveling Structural Information of Multi-Domain Nonribosomal Peptide Synthetases by Using Photo-Crosslinking Analysis with Genetic Code Expansion.- A Chemoenzymatic Approach to Investigate Cytochrome P450 Crosslinking in Glycopeptide Antibiotic Biosynthesis.- Cross-Linking of the Nonribosomal Peptide Synthetase Adenylation Domain with a Carrier Protein Using a Pantetheine-Type Probe.- A Practical Guideline to Engineering Non-Ribosomal Peptide Synthetases.- Probing Substrate-Loaded Carrier Proteins by Nuclear Magnetic Resonance.- Ribosomal Synthesis of Peptides Bearing Noncanonical Backbone Structures via Chemical posttranslational modifications.- Thioester Capture Strategy for the Identification of Nonribosomal Peptide and Polyketide Intermediates.- Chemical Labeling of Protein 4'-Phosphopantetheinylation in Surfactin-Producing Nonribosomal Peptide Synthetases.- Norine: Bioinformatics Methods and Tools for the Characterization of Newly Discovered NonRibosomal Peptides.
