Nonribosomal Peptide and Polyketide Biosynthesis

This volume presents an overview of nonribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) structure and function. It then continues with methods for the analysis of these pathways including conventional enzymological assays, contemporary mass spectrometric analysis techniques, specialized molecular biological approaches applicable to NRPSs and PKSs, and small molecule analysis tools tailored to this very special class of natural products, and concludes by examining bioinformatics tools for the analysis of these enzymes, pathways, and molecules. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Nonribosomal Peptide and Polyketide Biosynthesis: Methods and Protocols serves as a valuable reference for those experienced in studying NRPS and PKS enzymes, pathways, and natural products as well as a gateway for those just entering the field.

Includes cutting-edge methods and protocols for NPRS and PKS research Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras

Inhalt
Structural Biology of Nonribosomal Peptide Synthetases.- The Assembly Line Enzymology of Polyketide Biosynthesis.- Measurement of Nonribosomal Peptide Synthetase Adenylation Domain Activity Using a Continuous Hydroxylamine Release Assay.- Affinity Purification Method for the Identification of Nonribosomal Peptide Biosynthetic Enzymes Using a Synthetic Probe for Adenylation Domains.- Colorimetric Detection of the Adenylation Activity in Nonribosomal Peptide Synthetases.- Facile Synthetic Access to Glycopeptide Antibiotic Precursor Peptides for the Investigation of Cytochrome P450 Action in Glycopeptide Antibiotic Biosynthesis.- Reconstitution of Fungal Nonribosomal Peptide Synthetases in Yeast and In Vitro.- The Continuing Development of E. coli as a Heterologous Host for Complex Natural Product Biosynthesis.- Screening for Expressed Nonribosomal Peptide Synthetases and Polyketide Synthases Using LCMS/MS-Based Proteomics.- Enhancing Nonribosomal Peptide Biosynthesis in Filamentous Fungi.- In Situ Analysis of Bacterial Lipopeptide Antibiotics by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging.- Secondary Metabolic Pathway-Targeted Metabolomics.- Annotating and Interpreting Linear and Cyclic Peptide Tandem Mass Spectra.- Bioinformatics Tools for the Discovery of New Nonribosomal Peptides.- The Use of ClusterMine360 for the Analysis of Polyketide and Nonribosomal Peptide Biosynthetic Pathways.- Alignment-Free Methods for the Detection and Specificity Prediction of Adenylation Domains.- Characterization of Nonribosomal Peptide Synthetases with NRPSsp.