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</p><div><p><strong>
Chapter ONE
</strong> </p><p>1.0 INTRODUCTION AND LITERATURE REVIEW </p><p>1.1 Occurrence and Distribution</p><p>1.2 Assay Method</p><p>1.3 Methods of Purification</p><p>1.4 Structural properties of rhodanese</p><p>1.4.1 Molecular Weight</p><p>1.4.2 Amino Acid Composition</p><p>1.4.3 Amino Acid Sequence</p><p>1.4.4 Tertiary Structure of Rhodanese</p><p>1.4.5 Active site of Rhodanese</p><p>1.4.6 Mechanism of Action of rhodanese</p><p>1.5 Physical and Catalytic properties of rhodanese</p><p>1.5.1 Isoelectric point</p><p>1.5.2 Effect of pH</p><p>1.5.3 Effect of Temperature</p><p>1.5.4 Substrate specificity</p><p>1.5.5 Turnover number</p><p>1.5.6 Inhibition</p><p>1.5.7 Effect of Sulphyldryl reagent</p><p>1.6 Cloning of rhodanese gene</p><p>1.7 Biological importance of rhodanese</p><p>1.8 Background information</p><p>1.9 Specific Aims of research</p><p>1.10 Justification of research</p><p>
Chapter TWO
</p><p>2.0 MATERIALS AND METHODS </p><p>2.1 Materials</p><p>2.2 Methods</p><p>2.2.1 Preparation of Buffer and Reagents</p><p>2.2.1.1 Preparation of Borate buffer</p><p>2.2.1.2 Preparation of 15% formaldehyde from 40% formaldehyde</p><p>2.2.1.3 Preparation of sorbo reagent</p><p>2.2.1.4 Preparation of Bradford reagent</p><p>2.2.1.5 Preparation of 0.25M KCN</p><p>2.2.2 Enzyme source</p><p>2.2.3 Isolation and Purification of the enzyme</p><p>2.2.3.1 Homogenization</p><p>2.2.3.2 Ammonium sulphate precipitation</p><p>2.2.3.3 Reactive Blue-2-Agarose Affinity chromatography</p><p>2.2.4 Enzyme assay</p><p>2.2.5 Protein determination</p><p>2.2.6 Determination of kinetic parameters</p><p>2.2.7 Effect of pH on enzyme activity</p><p>2.2.8 Effect of Temperature on enzyme activity</p><p>2.2.9 Effect of Cations on enzyme</p><p>
Chapter THREE
</p><p>RESULTS </p><p>3.1 Purification of Rhodanese</p><p>3.2 Kinectic Parameters</p><p>3.3 Substrate Specificity</p><p>3.3.1 Effects of pH and temperature on enzyme activity</p><p>3.3.2 Effects of Cations</p><p><strong>
Chapter FOUR
</strong></p><p>4.0 Discussion and conclusion</p><p>4.1 Discussion</p><p>4.2 Conclusion</p><p>4.3 Recommendation</p><p><strong>REFERENCES.</strong></p><p></p></div><h3></h3><br>
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