Engineering Nonribosomal Peptide Synthesis by Directed Evolution and Module Reassembly PDF Download
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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: 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: Dan Robertson
Publisher: Elsevier
ISBN: 008049725X
Category : Science
Languages : en
Pages : 453
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Book Description
This MIE volume covers methods for a multitude of topics among which are computational methods, laboratory methods, enzyme optimization, binding proteins/antibodies, and screening technologies.Table of Contents-Methodology-Applications-Opzimization and Screening-Applications-Directed Evolution of Enzymatic Function-Applications-Evolution of Biosynthetic Pathways-Devices, Antibodies and Vaccines
Author: Nick J Westwood
Publisher: Royal Society of Chemistry
ISBN: 178801507X
Category : Science
Languages : en
Pages : 317
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Book Description
Synthetic chemistry plays a central role in many areas of chemical biology; utilising recent case studies, the goal of Chemical and Biological Synthesis is to highlight the full impact that the preparation of novel reagents can have in chemical biology. Covering the synthetic approaches that can be applied across the whole field of chemical biology, this book provides synthetic chemists with the broader context to which their work contributes and the biological questions that can be addressed through it. An ideal guide for postgraduate students and researchers in synthetic organic chemistry and chemical biology, Chemical and Biological Synthesis introduces synthetic techniques and methods to those who wish to incorporate synthesis for the first time in their biology-focused research programmes.
Author: Sung-Kun (Sean) Kim
Publisher: Frontiers Media SA
ISBN: 2889666859
Category : Science
Languages : en
Pages : 120
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Book Description
Author: Lutz Hilterhaus
Publisher: John Wiley & Sons
ISBN: 3527336699
Category : Science
Languages : en
Pages : 464
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Book Description
This reference book originates from the interdisciplinary research cooperation between academia and industry. In three distinct parts, latest results from basic research on stable enzymes are explained and brought into context with possible industrial applications. Downstream processing technology as well as biocatalytic and biotechnological production processes from global players display the enormous potential of biocatalysts. Application of "extreme" reaction conditions (i.e. unconventional, such as high temperature, pressure, and pH value) - biocatalysts are normally used within a well defined process window - leads to novel synthetic effects. Both novel enzyme systems and the synthetic routes in which they can be applied are made accessible to the reader. In addition, the complementary innovative process technology under unconventional conditions is highlighted by latest examples from biotech industry.
Author: Benjamin F. Cravatt
Publisher: Springer
ISBN: 3030111431
Category : Medical
Languages : en
Pages : 420
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Book Description
This volume provides a collection of contemporary perspectives on using activity-based protein profiling (ABPP) for biological discoveries in protein science, microbiology, and immunology. A common theme throughout is the special utility of ABPP to interrogate protein function and small-molecule interactions on a global scale in native biological systems. Each chapter showcases distinct advantages of ABPP applied to diverse protein classes and biological systems. As such, the book offers readers valuable insights into the basic principles of ABPP technology and how to apply this approach to biological questions ranging from the study of post-translational modifications to targeting bacterial effectors in host-pathogen interactions.
Author: Derek Barton
Publisher: Newnes
ISBN: 0080912834
Category : Science
Languages : en
Pages : 8492
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Book Description
Comprehensive Natural Products Chemistry
Author:
Publisher: Elsevier
ISBN: 0081026919
Category : Science
Languages : en
Pages : 4266
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Book Description
Comprehensive Natural Products III, Third Edition, Seven Volume Set updates and complements the previous two editions, including recent advances in cofactor chemistry, structural diversity of natural products and secondary metabolites, enzymes and enzyme mechanisms and new bioinformatics tools. Natural products research is a dynamic discipline at the intersection of chemistry and biology concerned with isolation, identification, structure elucidation, and chemical characteristics of naturally occurring compounds such as pheromones, carbohydrates, nucleic acids and enzymes. This book reviews the accumulated efforts of chemical and biological research to understand living organisms and their distinctive effects on health and medicine and to stimulate new ideas among the established natural products community. Provides readers with an in-depth review of current natural products research and a critical insight into the future direction of the field Bridges the gap in knowledge by covering developments in the field since the second edition published in 2010 Split into 7 sections on key topics to allow students, researchers and professionals to find relevant information quickly and easily Ensures that the knowledge within is easily understood by and applicable to a large audience
Author: Yuan Lu
Publisher: Springer Nature
ISBN: 3031412877
Category : Science
Languages : en
Pages : 154
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Book Description
This book reviews cell-free production systems, exploring the frontiers in cellular engineering and biotechnology. With contributions from experts in the field, the book offers a comprehensive and up-to-date account of the latest advancements and practical applications. The volume covers a diverse range of topics, beginning with an in-depth analysis of cell-free display techniques for protein evolution, shedding light on the methodologies used to engineer proteins for diverse purposes, followed by an examination of bottom-up synthetic biology employing cell-free protein synthesis. Additionally, it investigates the intricacies of the cell-free synthesis of metalloproteins, elucidating the unique properties and functionalities of these biologically important molecules. In this book, particular attention is given to the integration of cell-free production systems with droplet microfluidics, a pioneering approach that has revolutionized research activities in both academic and industrial settings. Readers will also discover the latest advancements in cell-free protein synthesis and immobilization, and find out more about the eCell technology, which combines cell-free protein synthesis with bio-sensing and remediation, revolutionizing critical areas of study in biotechnology. Together with the companion volume entitled “Cell-free Production: System Development”, both books highlight the research progresses on the basic and applied research of cell-free production systems in the last few years, and are invaluable resources for scholars, researchers, and bioengineers. This book also appeals to enthusiasts of synthetic biology.
Author:
Publisher:
ISBN:
Category : Dissertations, Academic
Languages : en
Pages : 362
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Book Description
