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Nonribosomal Peptide and Polyketide Biosynthesis

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Cover of 'Nonribosomal Peptide and Polyketide Biosynthesis'

Table of Contents

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    Book Overview
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    Chapter 1 Structural Biology of Nonribosomal Peptide Synthetases.
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    Chapter 2 The Assembly Line Enzymology of Polyketide Biosynthesis.
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    Chapter 3 Measurement of Nonribosomal Peptide Synthetase Adenylation Domain Activity Using a Continuous Hydroxylamine Release Assay
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    Chapter 4 Affinity Purification Method for the Identification of Nonribosomal Peptide Biosynthetic Enzymes Using a Synthetic Probe for Adenylation Domains
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    Chapter 5 Colorimetric Detection of the Adenylation Activity in Nonribosomal Peptide Synthetases
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    Chapter 6 Facile Synthetic Access to Glycopeptide Antibiotic Precursor Peptides for the Investigation of Cytochrome P450 Action in Glycopeptide Antibiotic Biosynthesis
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    Chapter 7 Reconstitution of Fungal Nonribosomal Peptide Synthetases in Yeast and In Vitro.
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    Chapter 8 The Continuing Development of E. coli as a Heterologous Host for Complex Natural Product Biosynthesis
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    Chapter 9 Screening for Expressed Nonribosomal Peptide Synthetases and Polyketide Synthases Using LC-MS/MS-Based Proteomics.
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    Chapter 10 Enhancing Nonribosomal Peptide Biosynthesis in Filamentous Fungi.
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    Chapter 11 In Situ Analysis of Bacterial Lipopeptide Antibiotics by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging.
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    Chapter 12 Secondary Metabolic Pathway-Targeted Metabolomics.
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    Chapter 13 Annotating and Interpreting Linear and Cyclic Peptide Tandem Mass Spectra.
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    Chapter 14 Bioinformatics Tools for the Discovery of New Nonribosomal Peptides
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    Chapter 15 The Use of ClusterMine360 for the Analysis of Polyketide and Nonribosomal Peptide Biosynthetic Pathways.
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    Chapter 16 Alignment-Free Methods for the Detection and Specificity Prediction of Adenylation Domains.
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    Chapter 17 Characterization of Nonribosomal Peptide Synthetases with NRPSsp.
Attention for Chapter 7: Reconstitution of Fungal Nonribosomal Peptide Synthetases in Yeast and In Vitro.
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Chapter title
Reconstitution of Fungal Nonribosomal Peptide Synthetases in Yeast and In Vitro.
Chapter number 7
Book title
Nonribosomal Peptide and Polyketide Biosynthesis
Published in
Methods in molecular biology, January 2016
DOI 10.1007/978-1-4939-3375-4_7
Pubmed ID
26831704
Book ISBNs
978-1-4939-3373-0, 978-1-4939-3375-4
Authors

Cacho, Ralph A, Tang, Yi, Ralph A. Cacho, Yi Tang, Cacho, Ralph A.

Abstract

The emergence of next-generation sequencing has provided new opportunities in the discovery of new nonribosomal peptides (NRPs) and NRP synthethases (NRPSs). However, there remain challenges for the characterization of these megasynthases. While genetic methods in native hosts are critical in elucidation of the function of fungal NRPS, in vitro assays of intact heterologously expressed proteins provide deeper mechanistic insights in NRPS enzymology. Our previous work in the study of NRPS takes advantage of Saccharomyces cerevisiae strain BJ5464-npgA as a robust and versatile platform for characterization of fungal NRPSs. Here we describe the use of yeast recombination strategies in S. cerevisiae for cloning of the NRPS coding sequence in 2μ-based expression vector; the use of affinity chromatography for purification of NRPS from the total S. cerevisiae soluble protein fraction; and strategies for reconstitution of NRPSs activities in vitro.

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X Demographics

The data shown below were collected from the profiles of 2 X users who shared this research output. Click here to find out more about how the information was compiled.
 Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 18 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Germany 2 11%
Unknown 16 89%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 7 39%
Researcher 4 22%
Lecturer 1 6%
Student > Bachelor 1 6%
Student > Master 1 6%
Other 0 0%
Unknown 4 22%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 7 39%
Agricultural and Biological Sciences 2 11%
Chemistry 2 11%
Medicine and Dentistry 1 6%
Chemical Engineering 1 6%
Other 0 0%
Unknown 5 28%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 03 February 2016.
All research outputs
#18,437,241
of 22,842,950 outputs
Outputs from Methods in molecular biology
#7,922
of 13,127 outputs
Outputs of similar age
#284,426
of 393,571 outputs
Outputs of similar age from Methods in molecular biology
#846
of 1,470 outputs
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