Purification and characterization of rhodanese from the liver of mango tilapia (sarotherodon galilaeus) found in erinle reservoir, south west, nigeria

• <p> </p><div><p><strong>

Chapter ONE

INTRODUCTION

• </strong>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; </p><p>
• 1.0&nbsp; INTRODUCTION AND LITERATURE REVIEW &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; </p><p>
• 1.1&nbsp; &nbsp; &nbsp; &nbsp; Occurrence and Distribution</p><p>
• 1.2&nbsp; &nbsp; &nbsp; &nbsp; Assay Method</p><p>
• 1.3&nbsp; &nbsp; &nbsp; Methods of Purification</p><p>
• 1.4&nbsp; &nbsp; &nbsp; Structural properties of rhodanese</p><p>1.
• 4.1&nbsp; &nbsp; &nbsp; &nbsp;Molecular Weight</p><p>1.
• 4.2&nbsp; &nbsp; &nbsp; Amino Acid Composition</p><p>1.
• 4.3&nbsp; &nbsp; &nbsp; Amino Acid Sequence</p><p>1.
• 4.4&nbsp; &nbsp; Tertiary Structure of Rhodanese</p><p>1.
• 4.5&nbsp; &nbsp; &nbsp; Active site of Rhodanese</p><p>1.
• 4.6&nbsp; &nbsp; Mechanism of Action of rhodanese</p><p>
• 1.5&nbsp; &nbsp; &nbsp; Physical and Catalytic properties of rhodanese</p><p>1.
• 5.1&nbsp; Isoelectric point</p><p>1.
• 5.2&nbsp; &nbsp; Effect of pH</p><p>1.
• 5.3&nbsp; &nbsp; Effect of Temperature</p><p>1.
• 5.4&nbsp; &nbsp; &nbsp; Substrate specificity</p><p>1.
• 5.5&nbsp; &nbsp; Turnover number</p><p>1.
• 5.6&nbsp; &nbsp; Inhibition</p><p>1.
• 5.7&nbsp; &nbsp; Effect of Sulphyldryl reagent</p><p>
• 1.6&nbsp; &nbsp; &nbsp;Cloning of rhodanese gene</p><p>
• 1.7&nbsp; &nbsp; Biological importance of rhodanese</p><p>
• 1.8&nbsp; &nbsp; Background information</p><p>
• 1.9&nbsp; &nbsp; Specific Aims of research</p><p>
• 1.10&nbsp; Justification of research</p><p>

Chapter TWO

LITERATURE REVIEW

• </p><p>
• 2.0&nbsp; &nbsp; &nbsp; &nbsp;MATERIALS AND METHODS &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; </p><p>
• 2.1&nbsp; &nbsp; &nbsp; &nbsp; Materials</p><p>
• 2.2&nbsp; &nbsp; &nbsp; &nbsp; &nbsp;Methods</p><p>2.
• 2.1&nbsp; &nbsp; &nbsp; Preparation of Buffer and Reagents</p><p>2.2.
• 1.1&nbsp; Preparation of Borate buffer</p><p>2.2.
• 1.2&nbsp; Preparation of 15% formaldehyde from 40% formaldehyde</p><p>2.2.
• 1.3&nbsp; Preparation of sorbo reagent</p><p>2.2.
• 1.4&nbsp; Preparation of Bradford reagent</p><p>2.2.
• 1.5&nbsp; Preparation of 0.25M KCN</p><p>2.
• 2.2&nbsp; &nbsp; &nbsp; Enzyme source</p><p>2.
• 2.3&nbsp; &nbsp;Isolation and Purification of the enzyme</p><p>2.2.
• 3.1&nbsp; Homogenization</p><p>2.2.
• 3.2&nbsp; Ammonium sulphate precipitation</p><p>2.2.
• 3.3&nbsp; Reactive Blue-2-Agarose Affinity chromatography</p><p>2.
• 2.4&nbsp; &nbsp; Enzyme assay</p><p>2.
• 2.5&nbsp; &nbsp; &nbsp;Protein determination</p><p>2.
• 2.6&nbsp; &nbsp; &nbsp; Determination of kinetic parameters</p><p>2.
• 2.7&nbsp; &nbsp; Effect of pH on enzyme activity</p><p>2.
• 2.8&nbsp; &nbsp; Effect of Temperature on enzyme activity</p><p>2.
• 2.9&nbsp; &nbsp; Effect of Cations on enzyme</p><p>

Chapter THREE

RESEARCH METHODOLOGY

• </p><p>RESULTS &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;</p><p>
• 3.1&nbsp; &nbsp; &nbsp; Purification of Rhodanese</p><p>
• 3.2&nbsp; &nbsp; &nbsp; Kinectic Parameters</p><p>
• 3.3&nbsp; &nbsp; &nbsp; Substrate Specificity</p><p>3.
• 3.1&nbsp; Effects of pH and temperature on enzyme activity</p><p>3.
• 3.2&nbsp; Effects of Cations</p><p><strong>

Chapter FOUR

DATA PRESENTATION AND ANALYSIS

• </strong></p><p>
• 4.0&nbsp; &nbsp; &nbsp; &nbsp; Discussion and conclusion</p><p>
• 4.1&nbsp; &nbsp; &nbsp; &nbsp; Discussion</p><p>
• 4.2&nbsp; &nbsp; &nbsp; &nbsp; Conclusion</p><p>
• 4.3&nbsp; &nbsp; &nbsp; &nbsp; &nbsp;Recommendation</p><p><strong>REFERENCES.</strong></p><p></p></div><h3></h3><br> <br><p></p>