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

Chapter ONE

INTRODUCTION

•                                                                                                                
• 1.0  INTRODUCTION AND LITERATURE REVIEW                                          
• 1.1        Occurrence and Distribution1.2         Assay Method1.3       Methods of Purification1.4       Structural properties of rhodanese1.
• 4.1       Molecular Weight1.
• 4.2      Amino Acid Composition1.
• 4.3      Amino Acid Sequence1.
• 4.4    Tertiary Structure of Rhodanese1.
• 4.5      Active site of Rhodanese1.
• 4.6    Mechanism of Action of rhodanese1.5       Physical and Catalytic properties of rhodanese1.
• 5.1  Isoelectric point1.
• 5.2    Effect of pH1.
• 5.3    Effect of Temperature1.
• 5.4      Substrate specificity1.
• 5.5    Turnover number1.
• 5.6    Inhibition1.
• 5.7    Effect of Sulphyldryl reagent1.6      Cloning of rhodanese gene1.7     Biological importance of rhodanese1.8     Background information1.9     Specific Aims of research1.10   Justification of researchCHAPTER TWO2.0        MATERIALS AND METHODS                                                          
• 2.1        Materials2.2          Methods2.
• 2.1      Preparation of Buffer and Reagents2.2.
• 1.1  Preparation of Borate buffer2.2.
• 1.2  Preparation of 15% formaldehyde from 40% formaldehyde2.2.
• 1.3  Preparation of sorbo reagent2.2.
• 1.4  Preparation of Bradford reagent2.2.
• 1.5  Preparation of 0.25M KCN2.
• 2.2      Enzyme source2.
• 2.3   Isolation and Purification of the enzyme2.2.
• 3.1  Homogenization2.2.
• 3.2  Ammonium sulphate precipitation2.2.
• 3.3  Reactive Blue-2-Agarose Affinity chromatography2.
• 2.4    Enzyme assay2.
• 2.5     Protein determination2.
• 2.6      Determination of kinetic parameters2.
• 2.7    Effect of pH on enzyme activity2.
• 2.8    Effect of Temperature on enzyme activity2.
• 2.9    Effect of Cations on enzymeCHAPTER THREERESULTS                                                                                                                
• 3.1      Purification of Rhodanese3.2       Kinectic Parameters3.3       Substrate Specificity3.
• 3.1  Effects of pH and temperature on enzyme activity3.
• 3.2  Effects of CationsCHAPTER FOUR4.0         Discussion and conclusion4.1         Discussion4.2         Conclusion4.3          RecommendationREFERENCES.