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.

When marketing managers and financial managers join forces within any business, the result can often be poor communication on financial criteria and goals. The risk of this situation occurring is inevitably present when those with different professional backgrounds and roles are working in accordance with their own norms. In his seminal 1956 paper on general systems theory, the economist Kenneth Boulding referred to the phenomenon of "specialised ears and generalised deafness", which can be seen to exist when marketing managers are financially illiterate or when financial managers lack the necessary insights to design, implement and operate accounting systems which are useful to marketing managers in carrying out their roles. It is increasingly difficult to attach credence to the idea of marketing managers who lack financial skills, or financial managers who fail to relate to the context in which marketing managers operate. Understanding the marketing/accounting interface is therefore important in generating emergent properties from the interaction of marketers and accountants whereby the whole is greater than the sum of the parts. The chapters in this volume seek to address this challenge. This book was originally published as a special issue of the Journal of Marketing Management.

The book highlights work from many different labs that taught us abnormal HDACs potentially contribute to the development or progression of many human diseases including immune dysfunctions, heart disease, cancer, memory impairment, aging, and metabolic disorders.

The aims of this volume are to highlight the tremendous pharmacological potential of protein kinase and protein phosphatase inhibitors, to provide a thorough overview of the most remarkable achievements in the field and to illustrate how beneficial these studies can be for the advancement of both basic knowledge on biological regulation and deregulation and for the clinical treatment of a wide spectrum of diseases. This goal is attained by contributions of leader investigators in the field, who address the issue from different angles.

This volume commemorates the 30th anniversary of the discovery in 1971 of the first DNA topoisomerases, i.e topoisomerase I in E. coli, then provisionally named omega' protein, by the pioneering work of Jim Wang at Harvard University, who has contributed an article to this volume: Reflections on an accidental discovery' of the enzyme. Many kinds of topoisomerase have since been found from type I through to type VI in a variety of organisms ranging from viruses to higher eukaryote mammals. The wide distribution of enzymes in various forms of life implies that the DNA topoisomerase is essential for life.

In the mid 80's type I and II enzymes were found to be the intracellular targets of a number of efficacious anticancer drugs such as doxorubicin, mitoxantrone, etoposide and camptothecin as a result of a continued efforts of many investigators, especially Leroy Liu and his collaborators at Johns Hopkins University. Readers will find a series of chapters written by researchers actively engaged in the expanding field of topoisomerase and their inhibitors. The series of chapters cover review articles on pharmacology and the molecular mechanism of topoisomerase I- and II-targeting anticancer drugs in mammals and in the yeast Saccharomyces cerevisiae, which has proved to be a superb model organism for studies of anticancer drugs. This volume compiles up-to-date information on the topoisomerase-targeting compounds in clinical and preclinical development as a useful and important reference book for students and researchers in the field of pharmacology, toxicology, oncology and molecular biology.

The second international symposium on Pectins and Pectinases was organized by Wageningen University and Research Centre and held in Rotterdam, May 6-10, 2001. This successful meeting was attended by around 130 participants from more than 20 countries representing almost all of the groups and industries working woridwide on pectins and pectinases. Following the first meeting on this subject held in December 1995, the symposium defInitely forms a platform for researchers and industries working in the fIeld, all within their own discipline and expertise. The symposium demanded a written account and this book is the resuit of that. It contains aIl keynote lectures and other oral presentations and provides an update about the current research. SignifIcant progress has been made in the last 5 years. This book provides an up-to-date insight into the research on pectin and pectic enzymes involved in their biosynthesis, degradation, modifIcation or utilization. The progress in the elucidation of the chemical structure of pectin and mode of action and 3-D structure of the pectin degrading enzymes allows us to identify and influence the functionality of pectins and pectic enzymes, both in vitra after isolation as weIl in the plants themselves (in planta). Other contributions deal with new applications of both pectin and pectin-degrading enzymes, while more and more attention is paid to health and nutritional aspects ofpectins. The book provides a 'state of the art' account for both beginners and experienced researchers of almost all disciplines of pectin research.

The lipases and phospholipases represent a diverse group of enzymes that are expressed in animals, plants, fungi, and bacteria. Their ubiquitous distribution among all species is a testament to the essential roles played by these enzymes in lipid storage, mobilization and metabolism, membrane homeostasis and remodeling, endocrine and immune functions, and signal tra- duction. In humans, lipases and phospholipases are also thought to contribute to complex diseases, such as atherosclerosis, obesity, arthritis, and cancer, as well as to single gene defects, such as Wolman's disease and Type I hyperlipoproteinemia. Enzymatically, the lipases are unique, since they hydrolyze substrates that are either insoluble, or only partly soluble, in aq- ous solvents; thus, enzyme catalysis takes place at a lipid-water interface. The interface comprises at least two, and often more, discrete bulk and s- face phases, in which the enzyme, substrate, and products oflipolysis disperse among these phases based on their physical properties. Furthermore, the d- tribution of these components changes continuously as lipolysis proceeds. Thus, the lipases and phospholipases are fundamentally different from any other enzyme because of the physical complexity of the environment in which catalysis occurs.

Recent years have seen a rapid increase in the use of enzymes as food processing tools, as an understanding of their means of control has improved. Since publication of the first edition of this book many new products have been commercially produced and the corresponding number of published papers has swollen. This second edition has been fully revised and updated to cover changes in the last five years. It continues to provide food technologists, chemists, biochemists and microbiologists with an authoritative, practical and detailed review of the subject.

Different aspects of biocatalysis are discussed with a view to obtaining products in a practical way using microorganisms and enzymes in catalytic amounts but as if they were organic reagents. A limited number of reactions catalysed by microbial reagents have been examinded according to the following classes: C-C bond formation, including the reaction catalyzed by aldolases, enantiomeric oxynitrilases, and decarboxylases. Oxidations promoted by microorganisms leading to Bayer-Williger-type products and hydroxylation at saturated carbon also includes the production of synthons derived from the microbial oxidation of substituted aromatics of wide synthetic applications. Reductions of carbonyl group with yeasts and other microorganisms, including the factor affecting the stereoselectivity when using all-cell organisms. Group transfer reactions describes synthetic achievements and theoretical aspects concerning the use of lipases, esterases and acylases. Industrial applications of microbial reagents in the large-scale production of fine chemicals offers validation to the basic research in the field.

The first systematic overview of this key technique since the early 1990s, this authoritative reference is the only handbook available to include all recent developments. The author draws on his wide-ranging experience in both academia and industry to systematically cover all types of enzyme immobilization methods, such as adsorption-based and covalent immobilization, as well as enzyme entrapment and encapsulation. Throughout, a careful review of materials and techniques for the generation of functional immobilized enzymes benefits both developers and users of carrier-bound enzymes.
A must for biotechnologists, biochemists and preparative chemists using enzymes in their daily work.

Key Building Blocks via Enzyme-Mediated Synthesis, by Thomas Fischer and J rg Pietruszka
*
Engineered Biosynthesis of Plant Polyketides: Structure-Based and Precursor-Directed Approach, by Ikuro Abe
*
Enzymatic and Chemo-Enzymatic Approaches Towards Natural and Non-Natural Alkaloids: Indoles, Isoquinolines, and Others, by Joachim St ckigt, Zhong Chen, and Martin Ruppert
*
Chemoenzymatic and Bioenzymatic Synthesis of Carbohydrate Containing Natural Products, by Bohdan Ostash, Xiaohui Yan, Victor Fedorenko, and Andreas Bechthold
*
Total (Bio)Synthesis: Strategies of Nature and of Chemists, by Alexandra A. Roberts, Katherine S. Ryan, Bradley S. Moore, and Tobias A.M. Gulder

Printing Peptide Arrays with a Complementary Metal Oxide Semiconductor Chip, by Felix F. Loeffler, Yun-Chien Cheng, Bastian Muenster, Jakob Striffler, Fanny C. Liu, F. Ralf Bischoff, Edgar Doersam, Frank Breitling, Alexander Nesterov-Mueller. Protein Engineering as a Tool for the Development of Novel Bio production Systems, by Uwe T. Bornscheuer. Compartmentalization and Metabolic Channeling for Multienzymatic Biosynthesis: Practical Strategies and Modeling Approaches, by U. Jandt, C. You, Y. H.-P. Zhang, A.-P. Zeng. Cell-Free Systems: Functional Modules for Synthetic and Chemical Biology, by Marlitt Stech, Andreas K. Brodel, Robert B. Quast, Rita Sachse, Stefan Kubick. New Bio production Systems: From Molecular Circuits to Novel Reactor Concepts in Cell-Free Biotechnology, by Steffen Rupp. Cell-free Biosystems in the Production of Electricity and Bioenergy, by Zhiguang Zhu, Tsz Kin Tam, Y.-H. Percival Zhang. In Vitro Multi enzymatic Reaction Systems for Biosynthesis, by Ines Ardao, Ee Taek Hwang, An-Ping Zeng. Directed Multistep Biocatalysis Using Tailored Permeabilized Cells, by Steffen Krauser, Christian Weyler, Lisa Katharina Blass, Elmar Heinzle."

Soil enzymes are one of the vital key mediators involved in nutrient recycling and the decomposition of organic matter and thereby in maintaining soil quality and fertility. This Soil Biology volume covers the various facets of soil enzymes, such as their functions, biochemical and microbiological properties and the factors affecting their activities. Enzymes in the rhizosphere, in forest soils, and in volcanic ash-derived soils are described. Soil enzymes covered include phosphohydrolases, lignocellulose-degrading enzymes, phenol oxidases, fungal oxidoreductases, keratinases, pectinases, xylanases, lipases and pectinases. Several chapters treat the soil enzymatic activities in the bioremediation of soils contaminated with pesticides and pollutants such as oil, chlorinated compounds, synthetic dyes and aromatic hydrocarbons. The role of soil enzymes as bioindicators is a further important topic addressed.

The critically acclaimed laboratory standard for more than forty years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with more than 300 volumes (all of them still in print), the series contains much material still relevant today--truly an essential publication for researchers in all fields of life sciences.

This detailed volume explores poly(ADP-ribose) polymerases (PARPs) in the biology of eukaryotes and their relevance to human health. Beginning with a section on the detection and quantification of poly(ADP-ribose) polymer (pADPr), the book continues by delving into the identification of protein targets, functional analysis, the poly(ADP-ribosyl)ating pathway in chromatin and genes expression, as well as the use of animal models and PARP1 inhibitor design and testing, and more. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Poly(ADP-Ribose) Polymerase: Methods and Protocols, Third Edition presents essential new and classical methods for studying the pADPr-pathway.

This volume provides up-to-date and novel techniques for various screening technologies currently used in metagenomics and related areas. Starting with DNA/RNA isolation from environmental samples, the book continues by delving into areas such as current methods used to isolate DNA and construct metagenomic libraries, establishment of metagenome libraries in non-E. coli hosts, and topics like function-driven mining of metagenomic DNA, screening and analyzing protocols for a wide array of different genes encoding enzymes, bacterial viruses and much more. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Metagenomics: Methods and Protocols, Third Edition provides a comprehensive collection of up-to-date metagenome protocols and tools for the recovery of many major types of biocatalysts and allows for the easy setup of these screens in microbiology laboratories.

Enzymatic Analysis: A Practical Guide is a multipurpose manual of laboratory methods. It offers a systematic scheme for the analysis of biological materials from the level of the whole organ down to the single cell and beyond. It is intended as a guide to the development of new methods, to the refinement of old ones, and to the adaptation in general of methods to almost any scale of sensitivity. As some may realize, the book is a sequel to A Flexible System of Enzymatic Analysis, originally published in 1972. The major changes, other than an appropriate interchange of authors, consist of a wholly new chapter of methods and protocols for measuring enzymes, the addition of 13 new entries in the metabolite chapter, and a much superior chapter on enzymatic cycling. With considerable nostalgia, we have switched from DPN and TPN to NAD and NADP nomenclature, which no doubt will make Otto Warburg turn over in his grave. The incentives for the methodology in this book came from the rigorous demands of quantitative histochemistry and cytochemistry. These demands are specificity, simplicity, flexibility, and, of course, sensitivity-all likewise desirable attributes of methods for other purposes. The specificity is provided by the use of enzyme methods. Simplicity is achieved by leading all reactions to a final pyridine nucleotide step.

High Pressure Effects in Molecular Biophysics and Enzymology is designed to acquaint biochemists, biophysicists, and graduate students with advances in the application of high pressure in connection with spectroscopy as a research tool in the study of biomolecules. The 23 chapters written by leading authorities present an overview of current approaches to the use of high pressure in research on enzyme kinetics, protein folding and structure, lipid bilayer structure and organization, lipid-protein interaction, and DNA structure. This important, timely volume is the first devoted exclusively to high-pressure effects in biochemistry and will be the definitive reference in its subject for the next several years.

Volume 39, devoted solely to the vital research area on molybdenum and tungsten and their role in biology, offers a comprehensive and timely account of this fascinating topic by 40 distinguished international authorities. With more than 2200 references to assist further research, Molybdenum and Tungsten: Their Roles in Biological Processes is an essential resource for scientists and students in many disciplines, including bioinorganic, inorganic, and coordination chemistry; biochemistry; biophysics; molecular biology; enzymology; pharmacology; physiology; clinical chemistry; nutrition; toxicology; and environmental sciences.

The objective of the "Springer Handbook of Enzymes" is to provide in concise form data on enzymes sufficiently well characterized. Data sheets are arranged in their EC-Number sequence. The volumes are arranged according to enzyme classes. Considerable progress has been made in enzymology since the publication of the first edition (published as "Enzyme Handbook"): many enzymes are newly classified or reclassified. In the 2nd edition each entry is correlated with references and one or more source organisms. New datafields are created: "application" and "engineering" (for the properties of enzymes where the sequence has been changed). Altogether the amount of data has doubled so that the 2nd edition will consist of 39 volumes plus synonym index. This collection is an indispensable source of information for researchers in biochemistry, biotechnology, organic and analytical chemistry, and food sciences.

This fully updated edition provides a series of methods for how best to assess functions of histone deacetylases and acetyltransferases. The disease-relevance of dysregulated protein deacetylation by overexpressed or aberrantly activated histone deacetylases has spurred an intense search for novel and improved inhibitors of these enzymes, as reflected in this collection. Expert contributors explore the generation and evaluation of novel histone deacetylase inhibitors and new and improved techniques to assess acetylation-dependent molecular mechanisms in vitro and in vivo. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, HDAC/HAT Function Assessment and Inhibitor Development: Methods and Protocols, Second Edition serves as an ideal guide for researchers seeking to further elucidate this vital area of study.

Medicinal chemistry is both science and art. The science of medicinal chemistry offers mankind one of its best hopes for improving the quality of life. The art of medicinal chemistry continues to challenge its practitioners with the need for both intuition and experience to discover new drugs. Hence sharing the experience of drug research is uniquely beneficial to the field of medicinal chemistry. Drug research requires interdisciplinary team-work at the interface between chemistry, biology and medicine. Therefore, the topic-related series Topics in Medicinal Chemistry covers all relevant aspects of drug research, e.g. pathobiochemistry of diseases, identification and validation of (emerging) drug targets, structural biology, drugability of targets, drug design approaches, chemogenomics, synthetic chemistry including combinatorial methods, bioorganic chemistry, natural compounds, high-throughput screening, pharmacological in vitro and in vivo investigations, drug-receptor interactions on the molecular level, structure-activity relationships, drug absorption, distribution, metabolism, elimination, toxicology and pharmacogenomics. In general, special volumes are edited by well known guest editors.

This Microbiology Monographs volume covers the latest advances in laccase applications in bioremediation and waste valorisation. The first three chapters provide a comprehensive introduction to fungal and bacterial laccases (the two most important enzyme groups from an application viewpoint) and their practical use in bioremediation and lignocellulosic waste valorisation. Subsequent chapters discuss possible combinations of laccases and further potentially collaborating enzymes, and offer in-depth insights into laccase immobilisation for wastewater treatment and environmental biosensor applications of laccases. Lastly, the book addresses the quest for enzymes with improved and better-fitting properties, covering laccase engineering by directed and computational evolution, and novel enzymes from extreme environments. As such, it is a fascinating read for microbiologists in both industry and academia.

Cytochrome P450: Structure, Mechanism, and Biochemistry, third edition is a revision of a review that summarizes the current state of research in the field of drug metabolism. The emphasis is on structure, mechanism, biochemistry, and regulation. Coverage is interdisciplinary, ranging from bioinorganic chemistry of cytochrome P450 to its relevance in human medicine. Each chapter provides an in-depth review of a given topic, but concentrates on advances of the last 10 years.