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.

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.

This enzymology textbook for graduate and advanced undergraduate students covers the syllabi of most universities where this subject is regularly taught. It focuses on the synchrony between the two broad mechanistic facets of enzymology: the chemical and the kinetic, and also highlights the synergy between enzyme structure and mechanism. Designed for self-study, it explains how to plan enzyme experiments and subsequently analyze the data collected. The book is divided into five major sections: 1] Introduction to enzymes, 2] Practical aspects, 3] Kinetic Mechanisms, 4] Chemical Mechanisms, and 5] Enzymology Frontiers. Individual concepts are treated as stand-alone chapters; readers can explore any single concept with minimal cross-referencing to the rest of the book. Further, complex approaches requiring specialized techniques and involved experimentation (beyond the reach of an average laboratory) are covered in theory with suitable references to guide readers. The book provides students, researchers and academics in the broad area of biology with a sound theoretical and practical knowledge of enzymes. It also caters to those who do not have a practicing enzymologist to teach them the subject.

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.

Principles of Enzyme Kinetics discusses the principles of enzyme kinetics at an intermediate level. It is primarily written for first-year research students in enzyme kinetics. The book is composed of 10 chapters. Chapter 1 provides the basic principles of enzyme kinetics with a brief discussion of dimensional analysis. Subsequent chapters cover topics on the essential characteristics of steady-state kinetics, temperature dependence, methods for deriving steady-state rate equations, and control of enzyme activity. Integrated rate equations, and introductions to the study of fast reactions and the statistical aspects of enzyme kinetics are provided as well. Chemists and biochemists will find the book invaluable.

This book was written with the purpose of providing a sound basis for the design of enzymatic reactions based on kinetic principles, but also to give an updated vision of the potentials and limitations of biocatalysis, especially with respect to recent app- cations in processes of organic synthesis. The ?rst ?ve chapters are structured in the form of a textbook, going from the basic principles of enzyme structure and fu- tion to reactor design for homogeneous systems with soluble enzymes and hete- geneous systems with immobilized enzymes. The last chapter of the book is divided into six sections that represent illustrative case studies of biocatalytic processes of industrial relevance or potential, written by experts in the respective ?elds. We sincerely hope that this book will represent an element in the toolbox of gr- uate students in applied biology and chemical and biochemical engineering and also of undergraduate students with formal training in organic chemistry, biochemistry, thermodynamics and chemical reaction kinetics. Beyond that, the book pretends also to illustrate the potential of biocatalytic processes with case studies in the ?eld of organic synthesis, which we hope will be of interest for the academia and prof- sionals involved in R&D&I. If some of our young readers are encouraged to engage or persevere in their work in biocatalysis this will certainly be our more precious reward.

Few scientists have the knowledge to perform the studies that are necessary to discover and characterize enzyme inhibitors, despite the vested interest the pharmaceutical industry has in this field. Beginning with the most basic principles pertaining to simple, one-substrate enzyme reactions and their inhibitors, and progressing to a thorough treatment of two-substrate enzymes, Kinetics of Enzyme Action: Essential Principles for Drug Hunters provides biochemists, medicinal chemists, and pharmaceutical scientists with numerous case study examples to outline the tools and techniques necessary to perform, understand, and interpret detailed kinetic studies for drug discovery.