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Author: Rebecca Schomer
Publisher:
ISBN:
Category :
Languages : en
Pages : 140
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Book Description
Natural products (NPs) are an important source for antibacterial, antifungal and anticancer drugs. One medically relevant class of NP, nonribosomal peptides (NRPs), are constructed by a conserved enzymology called nonribosomal peptide synthetases (NRPSs). Efforts to metabolically engineer next generation antibacterial NRPs has been slow due to an incomplete understanding of NRPS enzymology. The recent discovery that MbtH-like proteins (MLPs), a superfamily of short polypeptides with chaperone-like properties, are required for the solubility and activity of many NRPSs was a significant breakthrough in our ability to characterize NRPSs in vitro. While MLP-encoding genes are usually found within the NRPS-encoding gene cluster they influence, cross pathway MLP/NRPS interactions occur when more than one MLP is encoded in a genome. These interactions are complex and have been shown to be variable between both noncognate and cognate MLP/NRPS partners. Prior to this thesis, we did not have a detailed understanding of which aspects of the MLP are required for functional interaction with an NRPS. Using Enterobactin (ENT) biosynthesis from Escherichia coli as a model system, this thesis makes progress characterizing the role of the MbtH-like protein (MLP) in NRPS enzymology by dissecting MLP/NRPS interactions by two approaches. First, I probe the ability of noncognate MLPs to interact in vivo and in vitro with the MLP-dependent NRPS, EntF. Second, I target the cognate MLP, YbdZ, and perform a complete survey of the MLP residues involved in functional interactions with EntF. These analyses of this model MLP/NRPS pair is the most detailed to date and offers valuable insight into future directed evolution studies of MLPs and MLP-dependent NRPSs. Significantly, this work demonstrates the complexities of MLP/NRPS interactions and highlights challenges that must be addressed when engineering MLP-dependent NRPS.
Author: Rebecca Schomer
Publisher:
ISBN:
Category :
Languages : en
Pages : 140
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Book Description
Natural products (NPs) are an important source for antibacterial, antifungal and anticancer drugs. One medically relevant class of NP, nonribosomal peptides (NRPs), are constructed by a conserved enzymology called nonribosomal peptide synthetases (NRPSs). Efforts to metabolically engineer next generation antibacterial NRPs has been slow due to an incomplete understanding of NRPS enzymology. The recent discovery that MbtH-like proteins (MLPs), a superfamily of short polypeptides with chaperone-like properties, are required for the solubility and activity of many NRPSs was a significant breakthrough in our ability to characterize NRPSs in vitro. While MLP-encoding genes are usually found within the NRPS-encoding gene cluster they influence, cross pathway MLP/NRPS interactions occur when more than one MLP is encoded in a genome. These interactions are complex and have been shown to be variable between both noncognate and cognate MLP/NRPS partners. Prior to this thesis, we did not have a detailed understanding of which aspects of the MLP are required for functional interaction with an NRPS. Using Enterobactin (ENT) biosynthesis from Escherichia coli as a model system, this thesis makes progress characterizing the role of the MbtH-like protein (MLP) in NRPS enzymology by dissecting MLP/NRPS interactions by two approaches. First, I probe the ability of noncognate MLPs to interact in vivo and in vitro with the MLP-dependent NRPS, EntF. Second, I target the cognate MLP, YbdZ, and perform a complete survey of the MLP residues involved in functional interactions with EntF. These analyses of this model MLP/NRPS pair is the most detailed to date and offers valuable insight into future directed evolution studies of MLPs and MLP-dependent NRPSs. Significantly, this work demonstrates the complexities of MLP/NRPS interactions and highlights challenges that must be addressed when engineering MLP-dependent NRPS.
Author: Karla Jeanette Esquilin-Lebron
Publisher:
ISBN:
Category :
Languages : en
Pages : 0
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Book Description
An important group of bioactive natural products produced by bacteria and fungi are the nonribosomal peptides. The biosynthesis of these structurally diverse metabolites is derived by the modular enzymology of nonribosomal peptide synthetases (NRPSs). The modularity of NRPSs reside in a set of repeating catalytic domains that work together to recognize and incorporate aryl or amino acid precursors into the final product. The repetitive nature of NRPSs makes them perfect candidates for the rational design of these enzymes by the rearrangement of modules or domains to modify the nonribosomal peptide chemical scaffolds. Successes of NRPS combinatorial biosynthesis approaches have been scarce. Recent findings by the Thomas laboratory and others revealed that many NRPSs in bacteria require members of the MbtH-like proteins (MLP) superfamily for their solubility or function. MLP/NRPS interactions and their complexity are an important factor that the field has not addressed during combinatorial biosynthesis efforts. Proper pairing of MLP/NRPS systems might be the key to successful combinatorial biosynthesis efforts. In this thesis I propose two approaches to overcome the MLP challenge in combinatorial biosynthesis. First, the identification or evolution of a "universal" MLP that can act as a molecular tool and interact with multiple NRPSs. In Chapter 2, I characterized an orphan MLP, MXAN_3118, from M. xanthus DK1622 and identify it can naturally interact with at least seven cognate NRPS systems and the enterobactin system from E. coli. The ability of MXAN_3118 to naturally interact with multiple NRPSs suggests this enzyme is a candidate universal MLP that can potentially interact with MLP-dependent NRPSs from different systems. My second approach is to understand how we can change an MLP-dependent NRPS to be independent of the MLP. In Chapter 3, I studied the recently discovered enterobactin (ENT) system from yeast and identified that after acquisition of the biosynthetic gene cluster and further evolution the yeast system is MLP-independent. I hypothesize that the yeast ENT system lost the requirement for MLP from a combination of factors; including changes in the protein structure and the yeast cell physiology. The work in this thesis to overcome the MLP-dependence of NRPS systems, or MLP challenge, provided groundwork that can contribute to the future success of natural product engineering.
Author: Christopher T. Walsh
Publisher: Royal Society of Chemistry
ISBN: 1788010760
Category : Science
Languages : en
Pages : 787
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Book Description
This textbook describes the types of natural products, the biosynthetic pathways that enable the production of these molecules, and an update on the discovery of novel products in the post-genomic era.
Author: Robert A. Samson
Publisher:
ISBN: 9789070351533
Category : Electronic books
Languages : en
Pages : 260
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Book Description
Author: Christopher J Schofield
Publisher: Royal Society of Chemistry
ISBN: 1849739501
Category : Science
Languages : en
Pages : 508
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Book Description
Since the discovery of the first examples of 2-oxoglutarate-dependent oxygenase-catalysed reactions in the 1960s, a remarkably broad diversity of alternate reactions and substrates has been revealed, and extensive advances have been achieved in our understanding of the structures and catalytic mechanisms. These enzymes are important agrochemical targets and are being pursued as therapeutic targets for a wide range of diseases including cancer and anemia. This book provides a central source of information that summarizes the key features of the essential group of 2-oxoglutarate-dependent dioxygenases and related enzymes. Given the numerous recent advances and biomedical interest in the field, this book aims to unite the latest research for those already working in the field as well as to provide an introduction for those newly approaching the topic, and for those interested in translating the basic science into medicinal and agricultural benefits. The book begins with four broad chapters that highlight critical aspects, including an overview of possible catalytic reactions, structures and mechanisms. The following seventeen chapters focus on carefully selected topics, each written by leading experts in the area. Readers will find explanations of rapidly evolving research, from the chemistry of isopenicillin N synthase to the oxidation mechanism of 5-methylcytosine in DNA by ten-eleven-translocase oxygenases.
Author: Vladimir Havlicek
Publisher: John Wiley & Sons
ISBN: 1118466616
Category : Science
Languages : en
Pages : 634
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Book Description
This book highlights analytical chemistry instrumentation and practices applied to the analysis of natural products and their complex mixtures, describing techniques for isolating and characterizing natural products. • Applies analytical techniques to natural products research – an area of critical importance to drug discovery • Offers a one-stop shop for most analytical methods: x-ray diffraction, NMR analysis, mass spectrometry, and chemical genetics • Includes coverage of natural products basics and highlights antibacterial research, particularly important as efforts to combat drug resistance gain prominence • Covers instrumental techniques with enough detail for both current practitioners and beginning researchers
Author: Shahid Naeem
Publisher: Oxford University Press
ISBN: 0199547955
Category : Business & Economics
Languages : en
Pages : 387
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Book Description
The book starts by summarizing the development of the basic science and provides a meta-analysis that quantitatively tests several biodiversity and ecosystem functioning hypotheses.
Author: Dieter Söll
Publisher:
ISBN: 9781555818333
Category : Aminoacyl-tRNA synthetases
Languages : en
Pages : 572
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Book Description
Author: Graham F. Hatfull
Publisher: John Wiley & Sons
ISBN: 1683673425
Category : Science
Languages : en
Pages : 1428
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Book Description
A comprehensive collection of perspectives by experts in mycobacterial molecular biology Mycobacterium tuberculosis causes one in four avoidable deaths in the developing world and kills more adults than malaria, AIDS, and all tropical diseases combined. Tuberculosis was named a global health emergency by the World Health Organization, a distinction no other disease has received. Although the study of mycobacterial genetics has expanded dramatically, with new investigations into mycobacterial growth, replication, metabolism, physiology, drug susceptibility, and virulence, most of the problems in tuberculosis control that existed in 2000 remain today. Advances in our understanding of mycobacterial genetics have been reflected in exciting recent developments. New diagnostic approaches can identify drug resistance within a few hours, promising new drugs are progressing through the pipeline and into the clinic, and a range of newly developed vaccines are being evaluated. It is an exciting time as the fruits of 30 years of intensive genetic investigation are finally beginning to emerge. Written by leading experts in the field, Molecular Genetics of Mycobacteria, Second Edition, Discusses key areas of current research in mycobacterial genetics Explains the genetics of the physiology, metabolism, and drug sensitivities of M. tuberculosis Presents genetic approaches for manipulating M. tuberculosis This book is an invaluable resource for anyone interested in the molecular genetics and molecular biology of mycobacteria.
Author: Mamadou Daffé
Publisher:
ISBN: 9781555814687
Category : Bacterial cell walls
Languages : en
Pages : 0
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Book Description
Explains the unique characteristics that cause this large group of bacteria responsible for tuberculosis and leprosy to function differently; serves as a valuable reference for those working in the areas of biochemistry, genetics, genomics, and immunology.
