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Using Mass Spectrometry For Biochemical Studies On Enzymatic Domains From Polyketide Synthases

Author: Matthew Jenner
Publisher: Springer
ISBN: 3319327232
Size: 59.94 MB
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This thesis reports studies on the substrate specificity of crucial ketosynthase (KS) domains from trans-AT Polyketide Synthases (PKSs). Using a combination of electrospray ionisation-mass spectrometry (ESI-MS) and simple N-acetyl cysteamine (SNAC) substrate mimics, the specificity of a range of KS domains from the bacillaene and psymberin PKSs have been succsessfully studied with regard to the initial acylation step of KS-catalysis. In addition, the ability to alter the substrate tolerance of KS domains by simple point mutations in the active site has been demonstrated. A series of acyl-ACPs have been synthesised using a novel methodology and employed to probe the substrate specificity of both KS domains and the previously uncharcterised acyl hydrolase domain, PedC. KS-catalysed chain elongation reactions have also been conducted and monitored by ESI-MS/MS. All KS domains studied exhibited higher substrate specificity at the elongation step than in the preceeding acylation step. Furthermore, a mechanism of reversible acylation is proposed using the PsyA ACP1-KS1 di-domain. The findings in this thesis provide important insights into mechanisms of KS specificity and show that mutagenesis can be used to expand the repertoire of acceptable substrates for future PKS engineering.

Advances In Enzyme Biotechnology

Author: Pratyoosh Shukla
Publisher: Springer Science & Business Media
ISBN: 8132210948
Size: 13.73 MB
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Enzyme Technology is one the most promising disciplines in modern biotechnology. In this book, the applications of a wide variety of enzymes are highlighted. Current studies in enzyme technology are focused towards the discovery of novel enzymes (termed “bio-discovery” or “bio-prospecting”) and the identification and elucidation of novel pathways of these novel enzymes with emphasis on their industrial relevance. With the development of molecular techniques and other bioinformatics tools, the time to integrate this subject with other fields in the life sciences has arrived. A rapid expansion of the knowledge base in the field of enzyme biotechnology has occurred over the past few years. Much of this expansion has been driven by the bio-discovery of many new enzymes from a wide range of environments, some extreme in nature, followed by subsequent protein (enzyme) engineering. These enzymes have found a wide range of applications, ranging from bioremediation, bio-monitoring, biosensor development, bioconversion to biofuels and other biotechnologically important value-added products. Hydrolases constitute a major component of the global annual revenue generated by industrial enzymes and the emphasis has therefore been placed on these enzymes and their applications. With the immense interest of researchers active in this area, this book will serve to provide information on current aspects in this field of study. In the current edition, the contributions of many diversified topics towards establishing new directions of research in the area of enzyme biotechnology are described. This book serves to provide a unique source of information to undergraduates, post graduates and doctoral courses in microbiology and biotechnology along with allied life sciences. The present edition of the book covers all important areas of enzyme biotechnology i.e. the wide variety of enzymes in the field of enzyme biotechnology and their industrial applications, new methods and state-of-the-art information on modern methods of enzyme discovery. This book will act as good resource on most of the current facets of enzyme technology for all students engaged in bioengineering and biotechnology.

Molecular Markers In Mycology

Author: Bhim Pratap Singh
Publisher: Springer
ISBN: 3319341065
Size: 20.53 MB
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The Kingdom fungi encompass a massive diversity of taxa with wide-ranging ecologies, life cycles, and morphologies ranging from unicellular aquatic chytrids to large mushrooms. Before molecular methods came in existence, taxonomists considered this Kingdom to be a member of the plant kingdom due to certain life styles like immobility and growth habitats. Molecular markers (also known as DNA markers), facilitated a better alternative method over traditional morphological methods, employed for the identification, characterization, and to understand the evolution of fungi. The morphological methods used for identification are mainly dependent on spore color or microscopic features whereas molecular markers are based on DNA polymorphism in the genomic organization. Phylogenetic studies reported in last decade, based on molecular markers, have reshaped the classification system of Kingdom fungi, which divided into one subkingdom, seven phyla, and ten subphyla. Recent advances in molecular mycology have opened the way for researchers to identify and characterize novel fungal species from unique environments. Mycology is concerned with the systematic study of fungi, including their genetic and biochemical properties, their use to humans as a source of medicine and food, as well as their dangers, such as poisoning and infections. In the 21st century with the development of DNA sequencing technologies and phylogenetic analysis based on molecular markers, new insights into fungal taxonomy were provided. This book contains a thorough discussion of molecular characterization and detection of different groups of fungi by using PCR-based markers and provides a comprehensive view of the applications and uses of different molecular markers in molecular mycology. It also addresses the recent molecular markers employed to solve the problems of identification and discusses current approaches used in molecular characterization and detection of fungi.

Drug Discovery From Nature

Author: S. Grabley
Publisher: Springer Science & Business Media
ISBN: 9783540648444
Size: 19.36 MB
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This book is unique in covering the present status and future potential of natural products in drug discovery. It provides readers with recent information regarding the impact on drug discovery, development and strategies, technical and automation aspects, and methods based on biochemistry as well as molecular biology, highlighting compounds from natural sources. Special emphasis is placed on the various strategies to gain access to natural compounds and combinatorial approaches by making use of both synthetic and biological methods.

Genetic Engineering Of Plant Secondary Metabolism

Author: Brian E. Ellis
Publisher: Springer Science & Business Media
ISBN: 1461525446
Size: 76.45 MB
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In this volume of Recent Advances in Phytochmistry you will find a record of the pioneering attempts of plant biochemists and molecular biologists to modify the patterns of secondary metabolism in plants, as presented at the 33rd annual meeting of the Phytochemical Society of North America, in Asilomar, California, on June 27 -July I, 1993. The studies described here represent a marriage of the newest of technologies with one of the oldest human activities, exploitation of plant chemistry. They also represent the beginning of a new era of phytochemical research, an era that will undoubtedly begin to provide answers to some of the long-standing questions that have absorbed plant biochemists for the past century. There is, for instance, a common deflating experience to which every worker in the area of plant secondary metabolism can probably relate. After hearing about the latest research findings regarding some aspect of remarkable compound "X", someone in the audience finally directs the inevitable question at the hapless speaker. "Tell me, is anything known as to the biological role of compound "X" in the plant?" The answer, in most cases, must be "essentially nothing"! This is a frustrating scenario for both the speaker and the audience, since the very fact that a complex biosynthetic pathway remains encoded in a plant genome points to an associated selective advantage. The problem is that establishing the nature and scale of that advantage is a very complex task.

Complex Enzymes In Microbial Natural Product Biosynthesis Part B Polyketides Aminocoumarins And Carbohydrates

Author:
Publisher: Academic Press
ISBN: 9780080923369
Size: 39.68 MB
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Microbial natural products have been an important traditional source of valuable antibiotics and other drugs but interest in them waned in the 1990s when big pharma decided that their discovery was no longer cost-effective and concentrated instead on synthetic chemistry as a source of novel compounds, often with disappointing results. Moreover understanding the biosynthesis of complex natural products was frustratingly difficult. With the development of molecular genetic methods to isolate and manipulate the complex microbial enzymes that make natural products, unexpected chemistry has been revealed and interest in the compounds has again flowered. This two-volume treatment of the subject will showcase the most important chemical classes of complex natural products: the peptides, made by the assembly of short chains of amino acid subunits, and the polyketides, assembled from the joining of small carboxylic acids such as acetate and malonate. In both classes, variation in sub-unit structure, number and chemical modification leads to an almost infinite variety of final structures, accounting for the huge importance of the compounds in nature and medicine. * Gathers tried and tested methods and techniques from top players in the field. * Provides an extremely useful reference for the experienced research scientist. * Covers biosynthesis of Polyketides, Tarpenoids, Aminocoumarins and Crabohydrates

Springer Handbook Of Marine Biotechnology

Author: Se-Kwon Kim
Publisher: Springer
ISBN: 3642539718
Size: 17.64 MB
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This Springer Handbook provides, for the first time, a complete and consistent overview over the methods, applications, and products in the field of marine biotechnology. A large portion of the surface of the earth (ca. 70%) is covered by the oceans. More than 80% of the living organisms on the earth are found in aquatic ecosystems. The aquatic systems thus constitute a rich reservoir for various chemical materials and (bio-)chemical processes. Edited by a renowned expert with a longstanding experience, and including over 60 contributions from leading international scientists, the Springer Handbook of Marine Biotechnology is a major authoritative desk reference for everyone interested or working in the field of marine biotechnology and bioprocessing - from undergraduate and graduate students, over scientists and teachers, to professionals. Marine biotechnology is concerned with the study of biochemical materials and processes from marine sources, that play a vital role in the isolation of novel drugs, and to bring them to industrial and pharmaceutical development. Today, a multitude of bioprocess techniques is employed to isolate and produce marine natural compounds, novel biomaterials, or proteins and enzymes from marine organisms, and to bring them to applications as pharmaceuticals, cosmeceuticals or nutraceuticals, or for the production of bioenergy from marine sources. All these topics are addressed by the Springer Handbook of Marine Biotechnology. The book is divided into ten parts. Each part is consistently organized, so that the handbook provides a sound introduction to marine biotechnology - from historical backgrounds and the fundamentals, over the description of the methods and technology, to their applications - but it can also be used as a reference work. Key topics include: - Marine flora and fauna - Tools and methods in marine biotechnology - Marine genomics - Marine microbiology - Bioenergy and biofuels - Marine bioproducts in industrial applications - Marine bioproducts in medical and pharmaceutical applications - and many more...

Practical Gas Chromatography

Author: Katja Dettmer-Wilde
Publisher: Springer
ISBN: 3642546404
Size: 21.63 MB
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Gas chromatography continues to be one of the most widely used analytical techniques, since its applications today expand into fields such as biomarker research or metabolomics. This new practical textbook enables the reader to make full use of gas chromatography. Essential fundamentals and their implications for the practical work at the instrument are provided, as well as details on the instrumentation such as inlet systems, columns and detectors. Specialized techniques from all aspects of GC are introduced ranging from sample preparation, solvent-free injection techniques, and pyrolysis GC, to separation including fast GC and comprehensive GCxGC and finally detection, such as GC-MS and element-specific detection. Various fields of application such as enantiomer, food, flavor and fragrance analysis, physicochemical measurements, forensic toxicology, and clinical analysis are discussed as well as cutting-edge application in metabolomics is covered.

Biosynthesis And Molecular Genetics Of Fungal Secondary Metabolites

Author: Susanne Zeilinger
Publisher: Springer
ISBN: 1493925318
Size: 16.52 MB
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​Fungi produce many chemically diverse secondary metabolites whose biological roles largely remain elusive. Within the increasing number of sequenced fungal genomes several important genes involved in secondary metabolite formation have been identified. Most of these genes are clustered and their coordinated transcription is controlled in a complex way by both narrow pathway-specific regulators as well as broad global transcription factors responsive to environmental cues. In recent years it was discovered many of the newly identified gene clusters are silent under laboratory conditions suggesting that the biosynthetic potential of fungi is far from being exploited. Besides identifying novel bioactive metabolites from still unexplored sources, the activation of these gene clusters by several approaches may result in the discovery of new substances with antibiotic and pharmaceutical benefits. This book covers recent advances in the field of fungal secondary metabolisms ranging from methodologies to biological aspects and will include the latest knowledge on fungal molecular biology, genomics, and metabolomics. With the related volume by Professor Juan-Francisco Martin, where the most relevant and well-studied fungal secondary metabolites are compiled, this book provides a comprehensive overview of the state-of-the-art of research on fungal secondary metabolites.

Fundamentals Of Powder Diffraction And Structural Characterization Of Materials Second Edition

Author: Vitalij Pecharsky
Publisher: Springer Science & Business Media
ISBN: 0387095799
Size: 49.56 MB
Format: PDF
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A little over ?ve years have passed since the ?rst edition of this book appeared in print. Seems like an instant but also eternity, especially considering numerous developments in the hardware and software that have made it from the laboratory test beds into the real world of powder diffraction. This prompted a revision, which had to be beyond cosmetic limits. The book was, and remains focused on standard laboratory powder diffractometry. It is still meant to be used as a text for teaching students about the capabilities and limitations of the powder diffraction method. We also hope that it goes beyond a simple text, and therefore, is useful as a reference to practitioners of the technique. The original book had seven long chapters that may have made its use as a text - convenient. So the second edition is broken down into 25 shorter chapters. The ?rst ?fteen are concerned with the fundamentals of powder diffraction, which makes it much more logical, considering a typical 16-week long semester. The last ten ch- ters are concerned with practical examples of structure solution and re?nement, which were preserved from the ?rst edition and expanded by another example – R solving the crystal structure of Tylenol .