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Author: Michael Burkart
Publisher: Springer Nature
ISBN: 1071632140
Category : Science
Languages : en
Pages : 324
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Book Description
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.
Author: Michael Burkart
Publisher: Springer Nature
ISBN: 1071632140
Category : Science
Languages : en
Pages : 324
Get Book Here
Book Description
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.
Author: Philipp Stephan
Publisher:
ISBN:
Category :
Languages : de
Pages : 0
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Book Description
Over the last decades, discovery rates of new antibiotics have declined while the occurrence of multi-resistant pathogens has increased, leading towards a post-antibiotic era. Many current antibiotics are natural products or derivatives, known for their excellent bioactivities but complex structures that complicate synthetic production. Many antibiotics, like penicillin and vancomycin, are peptides produced by nonribosomal peptide synthetases (NRPS). These multi-modular enzyme complexes function like assembly lines, with each module adding a specific amino acid to the peptide through the biocatalytic activities of distinct domains. Targeted NRPS engineering to alter structures of the produced peptides has faced challenges, but directed evolution of NRPS domains seems to offer a promising solution. This thesis presents a straightforward method for the directed evolution of adenylation (A) domains using LC-MS/MS detection of dipeptides in cell lysates, demonstrating its power by engineering the GrsB1 A domain from gramicidin S biosynthesis to select for L-piperazic acid (Piz) instead of its native substrate L-Pro without reducing enzyme activity. The modified antibiotic, Piz-gramicidin S, showed improved bioactivity. Despite compatibility issues between NRPS domains, kinetic analyses revealed that A domain substrate preferences ultimately determine final product ratios. Additionally, the thesis proposes a novel DNA-based approach for NRPS engineering, using zinc finger domains that bind specific DNA sequences to simplify module rearrangement by making NRPSs DNA-templated. This method offers a more efficient alternative to traditional cloning techniques by replacing the handling of large NRPS genes with much shorter sequences of the zinc finger domains. Overall, this research advances the engineering of NRPSs for developing new antibiotics with better efficacy against resistant pathogens and highlights the usefulness of directed evolution in this context.
Author: Jennifer Fick Brannock
Publisher:
ISBN:
Category :
Languages : en
Pages : 184
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Book Description
Author: Farzaneh Pourmasoumi
Publisher:
ISBN:
Category :
Languages : de
Pages : 0
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Book Description
Nonribosomal peptide synthetases (NRPSs) are an important source of pharmaceutically valuable natural products, including antibiotics. In depth understanding of enzymatic interactions responsible for the biosynthesis of these compounds by NRPSs is essential for re-engineering of these proteins to create structural diversity. In this work, we establish tools that expand the understanding and engineering of nonribosomal peptide synthetases.
Author: Reto Daniel Zwahlen
Publisher:
ISBN: 9789403406732
Category :
Languages : en
Pages :
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Author: Andreas Tietze
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
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Author: Bradley S. Evans
Publisher:
ISBN: 9781493933754
Category : Enzymology
Languages : en
Pages : 281
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Book Description
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 successfulMethods in Molecular Biologyseries, 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 Protocolsserves 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.
Author: Daniel John Edwards
Publisher:
ISBN:
Category :
Languages : en
Pages : 644
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Book Description
Author: Michael Andrew Fischbach
Publisher:
ISBN:
Category : Biomolecules
Languages : en
Pages : 264
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Book Description
Author: Janik Kranz
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
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