Methods to Determine Enzymatic Activity

Methods to Determine Enzymatic Activity

Author: Alane Beatriz Vermelho

Publisher: Bentham Science Publishers

Published: 2013-11-22

Total Pages: 334

ISBN-13: 1608053008

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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.


Book Synopsis Methods to Determine Enzymatic Activity by : Alane Beatriz Vermelho

Download or read book Methods to Determine Enzymatic Activity written by Alane Beatriz Vermelho and published by Bentham Science Publishers. This book was released on 2013-11-22 with total page 334 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.


Practical Enzymology

Practical Enzymology

Author: Hans Bisswanger

Publisher: John Wiley & Sons

Published: 2019-11-06

Total Pages: 533

ISBN-13: 3527820809

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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.


Book Synopsis Practical Enzymology by : Hans Bisswanger

Download or read book Practical Enzymology written by Hans Bisswanger and published by John Wiley & Sons. This book was released on 2019-11-06 with total page 533 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.


HPLC in Enzymatic Analysis

HPLC in Enzymatic Analysis

Author: Edward F. Rossomando

Publisher: John Wiley & Sons

Published: 2009-09-25

Total Pages: 480

ISBN-13: 0470111003

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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.


Book Synopsis HPLC in Enzymatic Analysis by : Edward F. Rossomando

Download or read book HPLC in Enzymatic Analysis written by Edward F. Rossomando and published by John Wiley & Sons. This book was released on 2009-09-25 with total page 480 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.


Enzyme Assays

Enzyme Assays

Author: Robert Eisenthal

Publisher: Practical Approach

Published: 2002

Total Pages: 310

ISBN-13: 9780199638208

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Enzyme assays are among the most frequently performed procedures in biochemistry and are routinely used to estimate the amount of enzyme present in a cell or tissue, to follow the purification of an enzyme, or to determine the kinetic parameters of a system. The range of techniques used tomeasure the rate of an enzyme-catalysed reaction is limited only by the nature of the chemical change and the ingenuity of the investigator. This book describes the design and execution of enzyme assays, covering both general principles and specific chapters.Building upon the highly popular first edition, this book combines revised or rewritten chapters with entirely new contributions. Topics include experimental protocols covering photometric, radiometric, HPLC, and electrochemical assays, along with methods for determining enzyme assays after gelelectrophoresis. The theory underlying each method is outlined, together with a description of the instrumentation, sensitivity and sources of error. Also included are chapters on the principles of enzyme assay and kinetic studies; techniques for enzyme extraction; high- throughout screening;statistical analysis of enzyme kinetic data; and the determination of active site concentration.This second edition of Enzyme Assays will be valuable not only to biochemists, but to researchers in all areas of the life sciences.


Book Synopsis Enzyme Assays by : Robert Eisenthal

Download or read book Enzyme Assays written by Robert Eisenthal and published by Practical Approach. This book was released on 2002 with total page 310 pages. Available in PDF, EPUB and Kindle. Book excerpt: Enzyme assays are among the most frequently performed procedures in biochemistry and are routinely used to estimate the amount of enzyme present in a cell or tissue, to follow the purification of an enzyme, or to determine the kinetic parameters of a system. The range of techniques used tomeasure the rate of an enzyme-catalysed reaction is limited only by the nature of the chemical change and the ingenuity of the investigator. This book describes the design and execution of enzyme assays, covering both general principles and specific chapters.Building upon the highly popular first edition, this book combines revised or rewritten chapters with entirely new contributions. Topics include experimental protocols covering photometric, radiometric, HPLC, and electrochemical assays, along with methods for determining enzyme assays after gelelectrophoresis. The theory underlying each method is outlined, together with a description of the instrumentation, sensitivity and sources of error. Also included are chapters on the principles of enzyme assay and kinetic studies; techniques for enzyme extraction; high- throughout screening;statistical analysis of enzyme kinetic data; and the determination of active site concentration.This second edition of Enzyme Assays will be valuable not only to biochemists, but to researchers in all areas of the life sciences.


Fitting Models to Biological Data Using Linear and Nonlinear Regression

Fitting Models to Biological Data Using Linear and Nonlinear Regression

Author: Harvey Motulsky

Publisher: Oxford University Press

Published: 2004-05-27

Total Pages: 352

ISBN-13: 9780198038344

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Most biologists use nonlinear regression more than any other statistical technique, but there are very few places to learn about curve-fitting. This book, by the author of the very successful Intuitive Biostatistics, addresses this relatively focused need of an extraordinarily broad range of scientists.


Book Synopsis Fitting Models to Biological Data Using Linear and Nonlinear Regression by : Harvey Motulsky

Download or read book Fitting Models to Biological Data Using Linear and Nonlinear Regression written by Harvey Motulsky and published by Oxford University Press. This book was released on 2004-05-27 with total page 352 pages. Available in PDF, EPUB and Kindle. Book excerpt: Most biologists use nonlinear regression more than any other statistical technique, but there are very few places to learn about curve-fitting. This book, by the author of the very successful Intuitive Biostatistics, addresses this relatively focused need of an extraordinarily broad range of scientists.


Enzymes

Enzymes

Author: Robert A. Copeland

Publisher: John Wiley & Sons

Published: 2004-04-07

Total Pages: 416

ISBN-13: 0471461857

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Fully updated and expanded-a solid foundation for understandingexperimental enzymology. This practical, up-to-date survey is designed for a broadspectrum of biological and chemical scientists who are beginning todelve into modern enzymology. Enzymes, Second Editionexplains the structural complexities of proteins and enzymes andthe mechanisms by which enzymes perform their catalytic functions.The book provides illustrative examples from the contemporaryliterature to guide the reader through concepts and data analysisprocedures. Clear, well-written descriptions simplify the complexmathematical treatment of enzyme kinetic data, and numerouscitations at the end of each chapter enable the reader to accessthe primary literature and more in-depth treatments of specifictopics. This Second Edition of Enzymes: A Practical Introductionto Structure, Mechanism, and Data Analysis features refinedand expanded coverage of many concepts, while retaining theintroductory nature of the book. Important new featuresinclude: A new chapter on protein-ligand binding equilibria Expanded coverage of chemical mechanisms in enzyme catalysisand experimental measurements of enzyme activity Updated and refined discussions of enzyme inhibitors andmultiple substrate reactions Coverage of current practical applications to the study ofenzymology Supplemented with appendices providing contact information forsuppliers of reagents and equipment for enzyme studies, as well asa survey of useful Internet sites and computer software forenzymatic data analysis, Enzymes, Second Edition isthe ultimate practical guide for scientists and students inbiochemical, pharmaceutical, biotechnical, medicinal, andagricultural/food-related research.


Book Synopsis Enzymes by : Robert A. Copeland

Download or read book Enzymes written by Robert A. Copeland and published by John Wiley & Sons. This book was released on 2004-04-07 with total page 416 pages. Available in PDF, EPUB and Kindle. Book excerpt: Fully updated and expanded-a solid foundation for understandingexperimental enzymology. This practical, up-to-date survey is designed for a broadspectrum of biological and chemical scientists who are beginning todelve into modern enzymology. Enzymes, Second Editionexplains the structural complexities of proteins and enzymes andthe mechanisms by which enzymes perform their catalytic functions.The book provides illustrative examples from the contemporaryliterature to guide the reader through concepts and data analysisprocedures. Clear, well-written descriptions simplify the complexmathematical treatment of enzyme kinetic data, and numerouscitations at the end of each chapter enable the reader to accessthe primary literature and more in-depth treatments of specifictopics. This Second Edition of Enzymes: A Practical Introductionto Structure, Mechanism, and Data Analysis features refinedand expanded coverage of many concepts, while retaining theintroductory nature of the book. Important new featuresinclude: A new chapter on protein-ligand binding equilibria Expanded coverage of chemical mechanisms in enzyme catalysisand experimental measurements of enzyme activity Updated and refined discussions of enzyme inhibitors andmultiple substrate reactions Coverage of current practical applications to the study ofenzymology Supplemented with appendices providing contact information forsuppliers of reagents and equipment for enzyme studies, as well asa survey of useful Internet sites and computer software forenzymatic data analysis, Enzymes, Second Edition isthe ultimate practical guide for scientists and students inbiochemical, pharmaceutical, biotechnical, medicinal, andagricultural/food-related research.


Colorimetric Determination of Nitrate Plus Nitrite in Water by Enzymatic Reduction, Automated Discrete Analyzer Methods

Colorimetric Determination of Nitrate Plus Nitrite in Water by Enzymatic Reduction, Automated Discrete Analyzer Methods

Author: Charles J Patton

Publisher: CreateSpace

Published: 2014-06-16

Total Pages: 44

ISBN-13: 9781500223311

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This report documents work at the U.S. Geological Sur- vey (USGS) National Water Quality Laboratory (NWQL) to validate enzymatic reduction, colorimetric determinative meth- ods for nitrate + nitrite in filtered water by automated discrete analysis. In these standard- and low-level methods (USGS I-2547-11 and I-2548-11), nitrate is reduced to nitrite with nontoxic, soluble nitrate reductase rather than toxic, granular, copperized cadmium used in the longstanding USGS auto- mated continuous-flow analyzer methods I-2545-90 (NWQL laboratory code 1975) and I-2546-91 (NWQL laboratory code 1979). Colorimetric reagents used to determine resulting nitrite in aforementioned enzymatic- and cadmium-reduction meth- ods are identical. The enzyme used in these discrete analyzer methods, designated AtNaR2 by its manufacturer, is produced by recombinant expression of the nitrate reductase gene from wall cress (Arabidopsis thaliana) in the yeast Pichia pastoris. Unlike other commercially available nitrate reductases we evaluated, AtNaR2 maintains high activity at 37°C and is not inhibited by high-phenolic-content humic acids at reaction temperatures in the range of 20°C to 37°C. These previously unrecognized AtNaR2 characteristics are essential for success- ful performance of discrete analyzer nitrate + nitrite assays (henceforth, DA-AtNaR2) described here.


Book Synopsis Colorimetric Determination of Nitrate Plus Nitrite in Water by Enzymatic Reduction, Automated Discrete Analyzer Methods by : Charles J Patton

Download or read book Colorimetric Determination of Nitrate Plus Nitrite in Water by Enzymatic Reduction, Automated Discrete Analyzer Methods written by Charles J Patton and published by CreateSpace. This book was released on 2014-06-16 with total page 44 pages. Available in PDF, EPUB and Kindle. Book excerpt: This report documents work at the U.S. Geological Sur- vey (USGS) National Water Quality Laboratory (NWQL) to validate enzymatic reduction, colorimetric determinative meth- ods for nitrate + nitrite in filtered water by automated discrete analysis. In these standard- and low-level methods (USGS I-2547-11 and I-2548-11), nitrate is reduced to nitrite with nontoxic, soluble nitrate reductase rather than toxic, granular, copperized cadmium used in the longstanding USGS auto- mated continuous-flow analyzer methods I-2545-90 (NWQL laboratory code 1975) and I-2546-91 (NWQL laboratory code 1979). Colorimetric reagents used to determine resulting nitrite in aforementioned enzymatic- and cadmium-reduction meth- ods are identical. The enzyme used in these discrete analyzer methods, designated AtNaR2 by its manufacturer, is produced by recombinant expression of the nitrate reductase gene from wall cress (Arabidopsis thaliana) in the yeast Pichia pastoris. Unlike other commercially available nitrate reductases we evaluated, AtNaR2 maintains high activity at 37°C and is not inhibited by high-phenolic-content humic acids at reaction temperatures in the range of 20°C to 37°C. These previously unrecognized AtNaR2 characteristics are essential for success- ful performance of discrete analyzer nitrate + nitrite assays (henceforth, DA-AtNaR2) described here.


Mechanisms of Catalysis

Mechanisms of Catalysis

Author:

Publisher: Academic Press

Published: 1991-01-28

Total Pages: 471

ISBN-13: 0080865968

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The remarkable expansion of information leading to a deeper understanding of enzymes on the molecular level necessitated the development of this volume which not only introduces new topics to The Enzymes series but presents new information on some covered in Volume I and II of this edition.


Book Synopsis Mechanisms of Catalysis by :

Download or read book Mechanisms of Catalysis written by and published by Academic Press. This book was released on 1991-01-28 with total page 471 pages. Available in PDF, EPUB and Kindle. Book excerpt: The remarkable expansion of information leading to a deeper understanding of enzymes on the molecular level necessitated the development of this volume which not only introduces new topics to The Enzymes series but presents new information on some covered in Volume I and II of this edition.


Methods of Soil Enzymology

Methods of Soil Enzymology

Author: Richard P. Dick

Publisher: John Wiley & Sons

Published: 2020-01-22

Total Pages: 416

ISBN-13: 0891188541

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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.


Book Synopsis Methods of Soil Enzymology by : Richard P. Dick

Download or read book Methods of Soil Enzymology written by Richard P. Dick and published by John Wiley & Sons. This book was released on 2020-01-22 with total page 416 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.


Principles of Enzyme Kinetics

Principles of Enzyme Kinetics

Author: Athel Cornish-Bowden

Publisher: Elsevier

Published: 2014-05-20

Total Pages: 221

ISBN-13: 1483164675

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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.


Book Synopsis Principles of Enzyme Kinetics by : Athel Cornish-Bowden

Download or read book Principles of Enzyme Kinetics written by Athel Cornish-Bowden and published by Elsevier. This book was released on 2014-05-20 with total page 221 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.