Written in a logical, easy-to-understand manner, Experiments in the Purification and Characterization of Enzymes: A Laboratory Manual is an indispensable resource for both students and instructors in the fields of biochemistry, molecular biology, chemistry, pharmaceutical chemistry, and related molecular life sciences such as cell biology, neurosciences, and genetics.
In the first decade of the twenty-first century, enzymes and their multiple applications have played a critical role in the discovery and development of many new therapeutic agents.This book is a coordinated compilation of research expertise and current opinion uniquely focused on enzymes and their properties and applications.
Compiled by editors with a combined pharmaceutical experience of over sixty years, the text provides in-depth reviews of recent developments in selective topics on biosynthesis, biocatalysis, and chemical biology of enzymes as it applies to drug discovery, development, and manufacture.
The first in a multi-part series on enzymes, this volume features three sections:
New Approaches to Find and Modify Enzymes describes the emerging field of metagenomics, presents the practical applications of directed evolution to enzymes and pathways, and explores approaches for the discovery and design of biocatalysts
Biocatalytic Applications reviews specific applications of different reactions in producing active pharmaceutical ingredients and surveys recent developments employing enzymes in organic synthesis
Biosynthetic Applications goes over successful drug discoveries and developments by combinatorial biosynthesis and reviews the application of combinatorial biosynthesis among multiple compatible hosts
These timely discussions, which cover everything from chemical biology of enzymes, to the redesign of binding and catalytic specificities of enzymes, make this volume a valuable tool for keeping up to date. As such, it is an important read for researchers, students, and professors in the study of biotechnology, life sciences, biochemistry, enzymology, medicinal chemistry, and natural products.
The popular textbook Introduction to Enzyme and Coenzyme Chemistry has been thoroughly updated to include information on the most recent advances in our understanding of enzyme action, with additional recent examples from the literature used to illustrate key points. A major new feature is the inclusion of two-colour figures, and the addition of over 40 new figures of the active sites of enzymes discussed in the text, in order to illustrate the interplay between enzyme structure and function.
This new edition provides a concise but comprehensive account from the perspective of organic chemistry, what enzymes are, how they work, and how they catalyse many of the major classes of enzymatic reactions, and will continue to prove invaluable to both undergraduate and postgraduate students of organic, bio-organic and medicinal chemistry, chemical biology, biochemistry and biotechnology.
With its wide range of topics and long historical pedigree, Advances in Enzymology and Related Areas of Molecular Biology can be used not only by students and researchers in molecular biology, biochemistry, and enzymology, but also by any scientist interested in the discovery of an enzyme, its properties, and its applications.
Recently there has been much industrial interest in xylan and its hydrolytic enzymatic complex, as a supplement and for the manufacturing of food, drinks, textiles, pulps and paper, and ethanol; and in xylitol production as a fermentation substrate for the production of enzymes. This book describes xylan as a major component of plant hemicelluloses.
Fully updated to reflect advances made in the field over recent years, new chapters in the second edition look at the use of enzymes in the reduction of acrylamide, in fish processing and in non-bread cereal applications such as flour confectionery. Genetic modification of source organisms (GMO) has been used to improve yields of purer enzymes for some time now but the newer technology of protein engineering (PE) of enzymes has the potential to produce purer, more targeted products without unwanted side activities, and a chapter is also included on this important new topic. Authors have been selected not only for their practical working knowledge of enzymes but also for their infectious enthusiasm for the subject.
The book is aimed at food scientists and technologists, ingredients suppliers, geneticists, analytical chemists and quality assurance personnel.
The full potential of enzyme inhibitors and activators can only be appreciated after elucidating the details of their kinetic mechanisms of action exploring the entire range of physiologically significant reactant concentrations. This book highlights the wide spectrum of allosteric enzyme modification in physiological occurrences as well as in pharmacological and biotechnological applications that embrace simple and multiple enzyme-modifier interactions. The reader is guided in the journey through this still partly uncharted territory with the aid of mechanistically-oriented criteria aimed at showing the logical way towards the identification of a particular mechanism.
Activation enzyme-catalyzed functionalization reaction introduces a functional group to a lipophilic compound. Functionalization modifies many foreign compounds to form reactive intermediates capable of interacting with cellular components (proteins, DNA and lipids), leading to a variety of conditions for diseases. Functionalized compounds are further metabolized through detoxification enzyme-catalyzed reactions, which result in an increase in the solubility of parent compounds and an inactivation of metabolic intermediates, thus facilitating their excretion from the body. To minimize the exposure of potentially toxic metabolic intermediates, it is essential to keep them at a minimum level.
Extensive investigations have revealed that foreign compound-metabolizing enzymes exhibit genetic polymorphisms. Variations in their activities can produce different results as to the susceptibility to potential toxic effects. Moreover, the expressions of activation enzymes and detoxification enzymes are inducible. A number of chemical compounds are capable of acting as modulators for these two classes of enzymes. These findings have lead to the proposal of modulating metabolizing enzymes as a useful approach for human health benefits. Importantly, many of these chemical compounds are present in human daily diets.
There are many advances that have been made in the past decades towards the understanding of functions and implications of activation enzymes and detoxification enzymes. An organized, concise overview is needed for the readers who are initially exposed to this important subject, particularly for students and researchers in the areas of biomedical sciences, biochemistry, nutrition, pharmacology and chemistry. This book is intended to serve this purpose as an introduction to the subject. Furthermore, major topics in the book, excluding catalytic reactions and structural properties, may have interest to other readers who have knowledge of basic sciences and understanding enzyme related information.
The book discusses subjects associated with foreign compound metabolizing enzymes with emphasis on biochemical aspects, including lipophilic foreign compounds, catalytic properties, reactive intermediates, biomedical and biochemical effects, genetic polymorphisms, enzyme inducibility, enzyme modulation for health benefits, dietary related enzyme modulators, and structural characteristics of enzyme inducers.
All enzymes are remarkable since they have the ability to increase the rate of a chemical reaction, often by more than a billion fold. Allosteric enzymes are even more amazing because they have the additional ability to change their rate in response to cellular activators or inhibitors. This enables them to control the pathway in which they are the regulatory enzyme. Since the effector molecules represent the current status of the cell for a given metabolic pathway, this results in very responsive and balanced metabolic states, and makes it possible for cells and organisms to be appropriately dynamic, and responsive, in a changing environment. This book provides a logical introduction to the limits for enzyme function, as dictated by the factors that are the limits for life. This book presents a complete description of all the mechanisms used for changing enzyme activity. Eight enzymes are used as model systems, because they have been studied extensively. Wherever possible, the human form of the enzyme is used to illustrate the regulatory features.
While authors often emphasize the few enzymes that have the most remarkable catalytic rates, this survey of enzymes has led to the author’s appreciation of some important general conclusions:
1. Most enzymes are not exceptionally fast; they are always good enough for their specific catalytic step.
2. Although enzymes could always be much faster if they changed so as to bind their substrates more weakly, actual enzymes must be able to discriminate in favor of their special substrate, and therefore they have sacrificed speed to obtain better binding. This means that specific control of individual metabolic steps is more important than overall speed.
3. Results for many hundreds of enzymes establish that a lower limit for a normal catalytic activity is 1 s-1. Most enzymes have a catalytic rate between 10 and 300 s-1.
4. Allosteric regulation always results in a change in the enzyme's affinity for its substrate. Even V-type enzymes (named for their large change in catalytic velocity) always have a corresponding change in affinity for their substrate.
Thomas Traut has a PhD in molecular biology, and has studied enzymes since 1974. As a professor at the University of North Carolina at Chapel Hill he has focused on enzyme regulation, and taught advanced enzymology to graduate students. Important findings from his research helped to define the mechanism of allosteric control for dissociating enzymes. In this group the active form of the enzyme is normally oligomeric (trimer, tetramer, hexamer, etc.) and the dissociated subunit has little or no activity. With the solution of crystal structures for these enzymes by other laboratories, it became established that these dissociating enzymes had the active site at the interface of two adjacent subunits. Therefore, only the oligomer could have a complete active site. Since binding of the substrate, or of an effector, stabilizes two adjacent subunits in contact, this from of regulation is an efficient form of conformational control.
Additional studies helped to establish the correspondence between the subunit size of an enzyme and the number of independent ligand binding sites. Modules are the smallest units of folded protein structure. They are normally 3 - 7 kDa in size, and can bind one ligand. Catalytic sites, or regulatory sites, are frequently formed by two adjacent modules. The total size of a protein subunit then gives a good estimate of the number of such different ligand binding sites.
This is a revised edition of a very successful book, which appeals to both academic and industrial markets.Illustrates the organic mechanism associated with each enzyme-catalyzed reactionMakes the connection between organic reaction mechanisms and enzyme mechanismsCompiles the latest information about molecular mechanisms of enzyme reactionsAccompanied by clearly drawn structures, schemes, and figuresIncludes an extensive bibliography on enzyme mechanisms covering the last 30 yearsExplains how enzymes can accelerate the rates of chemical reactions with high specificityProvides approaches to the design of inhibitors of enzyme-catalyzed reactionsCategorizes the cofactors that are appropriate for catalyzing different classes of reactionsShows how chemical enzyme models are used for mechanistic studiesDescribes catalytic antibody design and mechanismIncludes problem sets and solutions for each chapter Written in an informal and didactic style