Enzymes

PrefaceI. IntroductionII. Enzyme Techniques Measurement of Velocity Classes of Methods Some Practical Points The General Handling of Enzymes The Study of an Enzyme The Specific and Molecular Activities Enzyme Affinities Methods of Quantitatively Following Enzyme Reactions (a) Spectrophotometric Methods (b) Manometric Methods (c) The Thunberg Method (d) Electrode Methods (e) Polarimetric Methods (f) Chromatographic Methods (g) Chemical EstimationsIII. Enzyme Isolation Importance of Enzyme Purification Methods of Purification (a) Test (b) General Procedure (c) Source of Enzyme (d) Extraction Fractionation Methods (a) Fractional Precipitation by Change of pH (b) Fractional Denaturation by Heating (c) Fractional Precipitation with Organic Solvents (d) Fractional Precipitation by Salts (e) Fractional Adsorption (f) Column Chromatography (g) Crystallization (h) Sequence of Fractionation Methods (i) Other Methods Criteria of Purity of Enzymes Pitfalls in Working with Pure EnzymesIV. Enzyme Kinetics The Importance of Enzyme Kinetics Factors Influencing Enzyme Reaction Velocity (A) Effect of Enzyme Concentration (B) Effect of Substrate Concentration (c) Effect of pH (D) Effect of TemperatureV. Enzyme Classification Enzyme Commission's Numbering System Rules for Systematic and Trivial Nomenclatures (i) General Rules (ii) Rules for Particular Classes of Enzymes Enzyme Reactions 1. Oxidoreductases 2. Transferases 3. Hydrolases 4. Lyases 5. Isomerases 6. LigasesVI. Enzyme Specificity The Investigation of Enzyme Specificity General Observations on Specificity Stereospecificity of Enzymes Illustrative Examples of Enzyme Specificity Dehydrogenases Glucose Oxidase Methyltransferases Transketolase and Transaldolase Glycosyltransferases Phosphotransferases Esterases Phosphatases Glycosidases Peptidases Lyases Aldose Mutarotase Enzymes with Substrates Containing Hydrocarbon Chains General ConclusionsVII. Enzyme Mechanisms Mechanism of Hydrogen-Transfer Reactions Alcohol Dehydrogenase Glyceraldehydephosphate Dehydrogenase Lipoamide Dehydrogenase Mechanism of Transferring (or Hydrolysing) Enzymes (a) Identification of the Bond Affected (b) Isotope Exchange Methods (c) The Optical Inversion Method (d) Inferences from Competition and Other Effects (e) Identification of Intermediate Complexes Mechanism of Lyases Fumarate Hydratase Aspartate Ammonia-Lyase Mechanism of Isomerases Epimerases Methylmalonyl-CoA Racemase Cis-trans Isomerases Ketol-Isomerases ¿-Isomerases Methylmalonyl-CoA Mutase Mechanism of Ligases Triple-Transfer Mechanism Ternary Complex Mechanisms Mechanism of Metalloenzymes Metal-Activated Peptidases Zinc-Containing Dehydrogenases Metalloflavoproteins Haemoprotein EnzymesVIII.