Contents & References of Enzymatic production of 1-butyl oleate ester using Rhizopus oryza cell system
List:
The first chapter. 1
1 Introduction. 2
The second chapter. 7
A review of the concepts and studies done. 7
2 Preface. 8
2.1 Enzyme. 8
2.2 Enzyme history. 9
2.3 Lipase enzyme 10
2.4 The difference between lipase enzyme and carboxylesterase 11
2.5 Reasons for increased attention of researchers to lipase enzyme 11
2.6 Lipase enzyme reactions 12
2.7 Characteristics of lipase enzyme 14
2.7.1 Structural properties (presence Cap on the active site) 14
2.7.2 Surface activation. 15
2.7.3 Substrate selectivity 16
2.7.4 Resistance to temperature increase and changes (pH) 19
2.8 Lipase enzyme production 20
2.8.1 Sources of lipase enzyme production 20
2.8.2 Comparison of bacterial and fungal lipases and their applications 22
2.8.3 Lipases of filamentous fungi. 23
2.8.4 Isolation of enzymes 24
2.8.5 Microorganism growth and enzyme induction. 26
2.9 Cell stabilization. 27
2.9.1 Comparing the advantages and disadvantages of enzyme and cell immobilization. 27
2.9.2 Use of enzyme or fixed cell. 29
2.9.3 cell stabilization methods. 30
2.9.4 Selection of retainer and method for cell stabilization. 36
2.9.5 The oozing mechanism and location of lipase in the cell of Rhizopus oryza and the effect of stabilization on its oozing. 42
2.10 Lipase enzyme activity measurement methods 45
2.10.1 Lipase enzyme dehydration activity measurement methods 45
2.10.2 Lipase enzyme synthetic activity measurement methods 46
2.11 Lipase enzyme applications 47
2.12 Ester synthesis reactions. 49
2.12.1 Parameters affecting the progress of ester synthesis reaction. 51
2.12.2 Synthesis of 1-butyl oleate ester. 54
The third chapter. 62
Materials and methods 62
3 Preface. 63
3.1 Chemicals. 63
3.2 Tools and devices used 64
3.3 Microorganism. 65
3.4 Description of experiments 66
3.4.1 Solid culture of microorganism. 66
3.4.2 Production of mushroom spore solution. 66
3.4.3 Microorganism liquid culture. 68
3.5 Preparation of cellular biocatalyst. 68
3.5.1 Loofah sponge as a cell holder. 68
3.5.2 Stabilization of Rhizopus oryza fungus and preparation of biocatalyst. 68
3.5.3 Determining the amount of cellular biocatalyst water. 70
3.6 Drawing the growth curve of the microorganism Rhizopus oryza in free form. 70
3.7 Activation of molecular sieves. 71
3.8 Turbidity measurement method of measuring free fatty acids. 71
3.8.1 Preparation of copper acetate-pyridine reagent solution. 72
3.8.2 Drawing the standard curve of free fatty acid absorption to measure esterification activity. 72
3.9 Enzyme specific activity assay. 73
3.9.1 Synthetic activity of enzyme system (cellular biocatalyst) - ester production. 74
3.9.2 Dehydration activity of enzyme system (cellular biocatalyst) 75
3.10 Synthesis reaction of 1-butyl oleate ester. 75
3.10.1 Analysis to determine the progress of the reaction. 76
3.10.2 Analysis of the manufactured product by gas chromatography-mass spectroscopy method. 76
3.11 Selection of the reaction system for the synthesis of 1-butyl oleate ester in the presence and absence of solvent. 77
3.11.1 Synthesis of 1-butyl oleate ester in the presence of hexane solvent. 77
3.11.2 Synthesis of 1-butyl oleate ester in the absence of solvent. 78
3.11.3 Comparison of the effect of mass transfer limitations inside the loofah piece, on the initial speed of the reaction in the presence of solvent and in the absence of solvent. 78
3.12 Optimizing reaction conditions for 1-butyl oleate ester synthesis in the presence of hexane solvent. 81
3.12.1 Investigating the effect of molar ratio of solvent to substrate on reaction efficiency and speed. 81
3.12.2 Investigating the effect of increasing the concentration of the alcoholic substrate. 81
3.12.3 Acidic substrate. 82
3.12.4 Investigating the effect of catalyst concentration on the efficiency and initial speed of the reaction. 82
3.12.1 Investigating the effect of removing water on reaction efficiency. 83
3.12.2 Investigating changes in efficiency in the successive use of cellular biocatalyst. 83
The fourth chapter. 84
Results and analyzes 84
4 Preface. 85
4.1 Growth curve of Rhizopus oryza 85
4.2 Investigation and comparison of cellular biocatalyst activity in free and stabilized form.86
4.2.1 The hydrolytic activity of the enzyme system (cellular biocatalyst) 86
4.2.2 The synthetic activity of the enzyme system (cellular biocatalyst) - ester production. 88
4.3 Selection of the reaction system for the synthesis of 1-butyl oleate ester in the presence and absence of solvent. 89
4.3.1 Synthesis of 1-butyl oleate ester in the presence of hexane solvent. 89
4.3.2 Synthesis of 1-butyl oleate ester in the absence of solvent. 91
4.3.3 Comparison of the internal mass transfer parameter of the loofah piece in the presence and absence of solvent. 92
4.4 Optimizing reaction conditions for the synthesis of 1-butyl oleate ester in the presence of hexane solvent. 97
4.4.1 Investigating the effect of molar ratio of solvent to substrate on reaction efficiency and speed. 97
4.4.2 Investigating the effect of catalyst concentration on the efficiency and initial speed of the reaction. 99
4.4.3 Investigating the effect of increasing the concentration of the alcoholic substrate. 100
4.4.4 Investigating the effect of increasing the concentration of the acidic substrate. 101
4.4.5 Investigating the effect of removing water on reaction efficiency. 103
4.4.6 Investigating the yield changes in the successive use of cellular biocatalyst. 105
Conclusion and suggestions. 107
5 Preface. 108
5.1 Conclusion. 108
5.2 Suggestions for future studies. 111
5.2.1 Biocatalyst. 111
5.2.2 Reaction substrate. 112
5.2.3 Reaction conditions. 112
5.2.4 Product. 113
Resources and references. 114
Appendix. 114
Source:
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