Evans B.S (Ed.) Nonribosomal Peptide and Polyketide Biosynthesis: Methods and Protocols

Humana Press, 2016 — 281p. — (Methods in Molecular Biology 1401) — ISBN: 978-1-4939-3375-4 (eBook), 978-1-4939-3373-0 (Hardcover).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.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 LC-MS/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