Methods of Enzymatic Analysis

Methods of Enzymatic Analysis focuses on the general progress in enzymology and in the special field of enzymatic analysis. This book explores the commercial production of biochemical reagents for analysis and explains the transition from the possible use of enzymatic analysis to its various applications in pure and applied biochemistry. Organized into four sections, this book starts with an overview of the basis of enzymatic analysis and provides general experimental guidelines for the techniques of measurement and for the disintegration of cells and tissues. This text then provides detailed instructions for the determination of substrates and assay of enzyme activities. Other chapters explore the practical aspects and information necessary for the application of reagents to enzymatic analysis, including sources, stability, and purity required. The final section describes the commercially available enzymes, coenzymes, substrates, and several less common reagents. Biochemists, biophysicists, researchers, and graduate students will find this book extremely useful.

Foreword

Preface

Contributors

Abbreviations

Section A: General

I. Principles of Enzymatic Analysis

II. Experimental Techniques

III. Cell and Tissue Disintegration

Section B: Estimation of Substrates

Preliminary Remarks

I. Substances Involved in Carbohydrate Metabolism

1. Poly-, Oligo- and Disaccharides

2. Monosaccharides and Derivatives

3. Three-Carbon Compounds

4. Two- and One-Carbon Compounds

II. Substances Involved in the Citric Acid Cycle

a) Citrate

b) Citrate and Isocitrate

c) α-Oxoglutarate

d) L-Malate

e) Oxaloacetate

f) Succinate

g) Fumarate

III. Substances Involved in Protein Metabolism

1. Proteins and Peptides

2. Amino Acids, Amines and Amides

IV. Substances Involved in the Metabolism of Fatty Acids, Lipids and Steroids

a) Polyunsaturated Fatty Acids

b) Lecithin

c) Acyl-CoA Derivatives

d) Acetoacetate

e) D-(—)-ß-Hydroxybutyrate

f) Hydrolysis of Steroid Conjugates

g) 20-Ketosteroids

h) Steroid Alcohols in Urine

V. Nucleosides, Purines and Pyrimidines, Coenzymes and Related Substances

1. Nucleosides, Purines and Pyrimidines

2. Coenzymes and Related Substances

VI. Other Substrates and Miscellaneous Substances

a) Organophosphorus Insecticides

b) DDT

c) Inorganic Peroxides

d) Nitrate

e) Magnesium in Plasma

f) Pyrophosphate

Section C: Measurement of Enzyme Activity

Preliminary Remarks

I. The Importance of the Measurement of Enzyme Activity

a) In Medicine

b) In Food Chemistry

c) In Botanical and Agricultural Chemistry

II. Methods

1. Aldolases

2. Dehydrogenases

3. Esterases

4. Proteases

5. Transaminases

6. Others

III. Histochemical Detection of Enzymes

1. Introduction (Aims of the Method)

2. General

3. Special Methods

Section D: Biochemical Reagents

I. Enzymes

II. Coenzymes and Substrates

III. Complete Reagent Kits

Tables

Conversion Table: Transmittance ⇌ Extinction

Atomic Weights

Index