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Methods of Enzymatic Analysis, Volume 2 reviews developments in the determination of enzyme activity, including advances in assay techniques. It discusses the principles on which measurements of enzymes are based, with each chapter including equations and each method consisting of the pipetting protocol. This volume is divided into four parts, each discussing a group of enzymes and their determination. Part I focuses on oxidoreductases, such as sorbitol dehydrogenase, lactate dehydrogenase, malate dehydrogenase, isocitrate dehydrogenase, 6-phosphogluconate dehydrogenase, xanthine oxidase, and glutamate dehydrogenase. Part II is concerned with transferases ranging from ornithine carbamoyltransferase and transamidinase to transketolase, transaldolase, UDP-glucuronyltransferase, glutamate-pyruvate transaminase, and phosphotransferases. Part III discusses hydrolases including esterases, glycoside hydrolases, peptidases, and proteinases, whereas Part IV looks at lyases, isomerases, and ligases, such as fructose-1, 6-diphosphate aldolase, 1-phosphofructoaldolase, glucosephosphate isomerase, and tetrahydrofolate formylase. This book is a valuable resource for biochemists as well as students and researchers working in the field of analytical biochemistry.
The use of High Performance Liquid Chromatography (HPLC) techniques in the study of enzymatic reactions has grown significantly since the publication of the first edition of this highly successful book: the role of enzymes in biological research has expanded; the application of HPLC and enzymes has extended to more disciplines; advances in separation techniques and instrumentation have increased the capability of HPLC; and the discovery of new enzymes has spawned new methods of analysis. High Performance Liquid Chromatography in Enzymatic Analysis, Second Edition addresses these developments in its coverage of the refinements of HPLC methods and their use in a wide range of laboratory applications. It offers the same practical approach found in the first edition, incorporates a wealth of new information into existing chapters, and adds new chapters to deal with new applications, including capillary electrophoresis, forensic chemistry, microdialysis, and the polymerase chain reaction. Topics include: * Application of HPLC to the assay of enzymatic activities * Concepts and principles of HPLC, including the latest technological advances * Concepts and principles of capillary electrophoresis (CE) * Strategy for design of an HPLC/CE system for assay of enzyme activity * Preparation of enzymatic activities from tissues and single cells * Analysis of enzymatic activities in body fluids, including chromatobiosis * HPLC for the identification of new enzymatic activities * Fundamentals of the polymerase chain reaction * HPLC in forensics * Survey of enzymatic activities assayed by the HPLC method, including many new categories * Multienzyme systems, including many new examples * HPLC in the analysis of contaminated food "It is the ability of HPLC to accomplish separations completely and rapidly that led to its original application to problems in the life sciences, particularly those related to purification. An analysis of the literature revealed that this technique was used primarily for the purification of small molecules, macromolecules such as peptides and proteins, and more recently, antibodies. This application to purification has all but dominated the use of the method, and there has been a plethora of books, symposia, and conferences on the use of HPLC for these purposes. However, it was only a matter of time before others began to look beyond and to explore the possibilities that result from the capacity to make separations quickly and efficiently." --from the preface to the First Edition Easy to read and full of practical advice and hundreds of diagrams and examples, High Performance Liquid Chromatography in Enzymatic Analysis, Second Edition is an invaluable resource for students, researchers, and laboratory workers in analytical chemistry and biochemistry, molecular biology and cell biology, and for anyone interested in keeping up with this fast-growing field.
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.
Methods to Determine Enzymatic Activity is a textbook about industrial enzymes. The book features definitions, classifications and applications of selected enzymes important in industry and in biotechnological processes. Analytical methods for these enzymes are also included in the text. The main objective of this textbook is to provide readers information focused on the current analysis methods of enzymatic activity at qualitative and quantitative levels. Each chapter is about one specific enzyme and contains information about its substrate and some biochemical properties. The methodologies are presented as an experimental protocol allowing interested readers to reproduce the experimental methods detailed within the textbook. These protocols contain the principle of the technique, materials, methods, and all steps necessary for the determination of enzyme activity and interpretation of results. Each methodology is illustrated with photos and schemes for a better and clear understanding. This book, therefore, uniquely brings modern analysis techniques of industrial enzymes in a single easy to understand volume. This textbook is suitable for undergraduate enzymology courses and advanced industrial biotechnology and microbiology courses.
Enzymatic Plastic Degradation, Volume 648 in the Methods in Enzymology series, continues the legacy of this premier serial with chapters authored by leaders in the field. Chapters in this latest release include Evaluating plastic pollution and environmental degradation, Assessment methods for microplastic pollution in the oceans and fresh water, Exploring microbial consortia from various environments for plastic degradation, Characterization of filamentous fungi for attack on synthetic polymers via biological Fenton chemistry, Synthesis of radioactive-labeled nanoplastics for assaying the environmental (microbial) PS degradation, Exploring metagenome for plastic degrading enzymes, Cutinases from thermophilic bacteria (actinomycetes): from identification to functional and structural characterization, and much more. - Provides the authority and expertise of leading contributors from an international board of authors - Presents the latest release in the Methods in Enzymology series - Covers the latest research and technologies in enzymatic plastic degradation
A practice-oriented guide to assaying more than 100 of the most important enzymes, complete with the theoretical background and specific protocols for immediate use in the biochemical laboratory. Now expanded with a new section on metal ion determination.
Methods of Soil Enzymology provides the first comprehensive set of vetted methods for studying enzymes in soils. Readers will especially benefit from the step-by-step explanation of the lab procedures, as well as background information for using these methods effectively and analyzing data. Main topics include activity assays, enzyme extraction, and synthetic enzyme complexes. Each method covered includes background informaton, step-by-step descriptions of the procedure, and special comments regarding nuances, pitfalls, and interpretation of the method. Learn the latest research methods, including enzyme extraction methods and procedures for creating synthetic enzyme complexes, as well as the newest ways to use small-scale and high-throughput methods for enzyme activity assays. Written for the researcher, but welcoming to those new to soil enzymology, the introduction includes conceptual information to orient those who are not familiar with these methods but want to use them. In the tradition of SSSA methods books, Methods of Soil Enzymology features a comprehensive approach with a focus on ease of use.
Practical Enzyme Kinetics provides a practical how-to guide for beginning students, technicians, and non-specialists for evaluating enzyme kinetics using common software packages to perform easy enzymatic analyses.