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Natural Product Biosynthesis by Microorganisms and Plants Part B - 1st Edition - ISBN: 9780123942913, 9780123946287

Natural Product Biosynthesis by Microorganisms and Plants Part B, Volume 516

1st Edition

Serial Volume Editor: David Hopwood
Hardcover ISBN: 9780123942913
eBook ISBN: 9780123946287
Imprint: Academic Press
Published Date: 1st October 2012
Page Count: 462
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Table of Contents

Chapter One. In Vivo Production of Thiopeptide Variants

1 Introduction

2 Generation of Thiopeptide Variants


Chapter Two. Microviridin Biosynthesis

1 Introduction

2 Reconstitution In Vitro of the Cyclization Reactions of the Microviridin Biosynthetic Pathway from P. agardhii

3 Influence of Leader Peptide on the Cyclization Reactions

4 Summary


Chapter Three. Cyclotide Isolation and Characterization

1 Introduction

2 Isolation of Cyclotides from Plant Tissues

3 Isolation of Nucleic Acids Encoding Cyclotide Precursors

4 Mass Spectrometric Detection and Characterization of Cyclotides

5 Structural Analysis of Cyclotides

6 Membranolytic Assays of Cyclotides

7 Summary


Chapter Four. Ribosomally Encoded Cyclic Peptide Toxins from Mushrooms

1 Introduction

2 Detection and Purification of Amanita Cyclic Peptide Toxins

3 Identification of Cyclic Peptide Genes in Amanita and Other Fungi

4 Purification and Assay of Prolyl Oligopeptidase (POP) from Mushrooms

5 Immunodetection of POP in Mushroom Tissues

6 Summary


Chapter Five. The Pictet–Spengler Mechanism Involved in the Biosynthesis of Tetrahydroisoquinoline Antitumor Antibiotics

1 Introduction

2 Biosynthetic Analysis of THIQ Core Scaffold Assembly

3 Reaction Mechanism for Constructing a Pentacyclic THIQ Core Skeleton Catalyzed by a Novel Function for a Single NRPS Module

4 Comparison Between Typical Peptide Synthesis and the Pictet–Spengler Reaction Catalyzed by NRPS Machinery

5 Reaction Profiles Affected by the Chain Length of the Acyl Moiety

6 Multiple Roles for the Fatty Acyl Chain During the Maturation Process in Late-Stage Biosynthesis

7 Methods for Substrate Synthesis, Enzyme Preparation, and Assay Conditions

8 Conclusion


Chapter Six. Discovery and Biosynthesis of Phosphonate and Phosphinate Natural Products

1 Introduction

2 Spectrum of Phosphonate Natural Products: Structures and Bioactivities

3 Biosynthetic Pathways

4 Screening Bacteria for Phosphonate Producing Gene Clusters

5 Purification of Phosphonates/Phosphinates

6 Structural Elucidation of Phosphonates

7 A Wealth of Unusual Biosynthetic Enzymes

8 Conclusion and Outlook


Chapter Seven. RlmN and AtsB as Models for the Overproduction and Characterization of Radical SAM Proteins

1 Introduction

2 RlmN as a Model for Gene Expression and Overproduction of RS Proteins

3 RlmN as a Model for Purification of RS Proteins

4 Overproduction and Purification of E. coli Flavodoxin and Flavodoxin Reductase

5 AtsB as a Model for Determination of Configuration and Stoichiometry of Iron–Sulfur Clusters in RS Enzymes

6 Activity Determination of RS Enzymes


Chapter Eight. Fe(II)-Dependent, Uridine-5′-Monophosphate α-Ketoglutarate Dioxygenases in the Synthesis of 5′-Modified Nucleosides

1 Introduction

2 Methods

3 Summary


Chapter Nine. Tailoring Reactions Catalyzed by Heme-Dependent Enzymes

1 Introduction

2 Overexpression, Purification, and Characterization of the Novel Nitrating CYP TxtE

3 Protein Characterization Using UV–vis Spectroscopy

4 Ligand and Substrate Binding

5 Summary


Chapter Ten. Oxidative Tailoring Reactions Catalyzed by Nonheme Iron-Dependent Enzymes

1 Introduction

2 Examples of Tailoring Reactions Catalyzed by Nonheme Iron- and Oxygen-Dependent Enzymes

3 Heterologous Expression and Functional Investigation of the S. coelicolor redH and redG Genes in Streptomyces albus

4 Conclusions


Chapter Eleven. The Rare Fluorinated Natural Products and Biotechnological Prospects for Fluorine Enzymology

1 Introduction

2 Production of Fluoroacetate and 4-Fluorothreonine from S. cattleya

3 Enzyme 1: Fluorinase

4 Enzyme 2: Purine Nucleoside Phosphorylase

5 Enzyme 3: Isomerase

6 Enzyme 4: Aldolase

7 Enzyme 5: Transaldolase (Pyridoxal Phosphate)

8 Enzyme 6: Aldehyde Dehydrogenase

9 Generation of a Novel Fluorometabolite by Genetic Engineering

10 Summary


Chapter Twelve. Enzymatic Chlorination and Bromination

1 Introduction

2 FADH2-Dependent Halogenases

3 Characterization of FADH2-Dependent Halogenases

4 Nonheme Iron, α-Ketoglutarate- and O2-Dependent Halogenases


Chapter Thirteen. Prenyltransferases of the Dimethylallyltryptophan Synthase Superfamily

1 Introduction

2 Spore Preparation

3 Cultivation of Fungi

4 RNA and DNA Isolation and cDNA Synthesis

5 Gene Cloning

6 Protein Overproduction

7 Preparation of Stock Solutions for Enzyme Assays

8 Biochemical Characterization of Prenyltransferases

9 Chemoenzymatic Synthesis of Prenylated Compounds


11 Summary


Chapter Fourteen. Serine Carboxypeptidase-Like Acyltransferases from Plants

1 Introduction

2 Methods in SCPL Acyltransferase Research

3 Summary


Chapter Fifteen. 4-Methylideneimidazole-5-One-Containing Aminomutases in Enediyne Biosynthesis

1 Introduction

2 Methods

3 Conclusion


Chapter Sixteen. Tailoring Enzymes Acting on Carrier Protein-Tethered Substrates in Natural Product Biosynthesis

1 Introduction

2 Methods

3 Conclusion


Chapter Seventeen. High-Throughput Colorimetric Assays for Nucleotide Sugar Formation and Glycosyl Transfer

1 Introduction

2 Purification of the Glycosyltransferase OleD

3 Colorimetric Assay for NDP-Sugar Formation

4 Coupled Reactions for Glycosyl Transfer

5 High-Throughput Colorimetric Coupled Assay for Glycosyl Transfer

6 Summary


Author Index

Subject Index


This new volume of Methods in Enzymology continues the legacy of this premier serial by containing quality chapters authored by leaders in the field. The second of 3 volumes covering Natural product biosynthesis by microorganisms and plants.

Key Features

  • This new volume continues the legacy of this premier serial
  • Contains quality chapters authored by leaders in the field
  • The second of 3 volumes it has chapters on such topics as biological chlorination, bromination and iodination, and phylogenetic approaches to natural product structure prediction


Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists


No. of pages:
© Academic Press 2012
1st October 2012
Academic Press
Hardcover ISBN:
eBook ISBN:

Ratings and Reviews

About the Serial Volume Editor

David Hopwood

Affiliations and Expertise

John Innes Centre and AFRC Institute of Plant Science Research, Norwich, UK