1.1.2 Existing technologies for the removal of micropollutants . . . 20

Contents

1 Introduction 19

1.1 Scope of the work . . . 19

1.1.1 Generalities . . . 19

1.1.2 Existing technologies for the removal of micropollutants . . . 20

1.1.3 The remarkable potential of lignolytic enzymes . . . 21

1.2 Objectives and strategy . . . 23

1.2.1 Objectives . . . 23

1.2.2 Strategy . . . 23

2 Products formation from phenolic compounds removal by laccases: a re- view 25 2.1 Abstract . . . 25

2.2 Abbreviations . . . 26

2.3 Keywords . . . 26

2.4 Introduction . . . 26

2.5 Laccases as a green catalyst . . . 27

2.6 Degradation mechanisms and product identification for the main studied mi- cropollutants . . . 29

2.7 Degradation mechanisms and product identification for less studied micropol- lutants . . . 37

2.7.1 Pharmaceuticals and personal care products . . . 37

2.7.2 Polycyclic aromatic hydrocarbons . . . 37

2.8 Degradation mechanisms and produc identification for other contaminants . . 38

2.8.1 Dyes . . . 38

2.8.2 Olive oil mill wastewater . . . 39

2.8.3 Substituted phenols . . . 40

2.9 Conclusion . . . 41

2.10 Acknowledgment . . . 44

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CONTENTS

3 Selection of new supports for laccases immobilization 45

3.1 Introduction . . . 45

3.2 Supports selection . . . 47

3.3 Materials and Methods . . . 47

3.3.1 Support characterization . . . 47

3.3.2 Laccases immobilization . . . 48

3.3.3 Packed-bed characterization . . . 50

3.4 Results and discussion . . . 53

3.4.1 Support characterization . . . 53

3.4.2 Laccases immobilization . . . 58

3.4.3 Packed-bed characterization . . . 63

3.5 Conclusion . . . 70

4 Insoluble reaction products from bisphenol A removal by laccases: kinetics and characterization 73 4.1 Introduction . . . 73

4.2 Kinetic study . . . 74

4.2.1 Material and Methods . . . 74

4.2.2 Results and discussion . . . 79

4.3 Kinetic modeling . . . 85

4.3.1 Model development . . . 85

4.3.2 Validation on experimental data . . . 87

4.4 Characterization . . . 88

4.4.1 Differential scanning calorimetry (DSC) . . . 88

4.4.2 Gel permeation chromatography (GPC) . . . 97

4.4.3 Fourier transform infrared spectroscopy (FTIR) . . . 99

4.5 Conclusion . . . 101

5 Soluble reaction products from bisphenol A removal by laccases: kinetics and characterization 103 5.1 Introduction . . . 103

5.2 Material and Methods . . . 104

5.2.1 Experimental configurations and sample acquisition . . . 104

5.2.2 LC/MS analytical method . . . 105

5.2.3 BPA monitoring . . . 107

5.2.4 BPA soluble reaction products screening and monitoring . . . 107

5.3 Results and discussion . . . 109

5.3.1 BPA removal kinetics . . . 109

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CONTENTS

5.3.2 BPA soluble reaction products isolation and identification . . . 113

5.3.3 BPA soluble reaction products formation kinetics: batch reactor and free enzymes . . . 118

5.3.4 BPA soluble reaction products formation kinetics: batch reactor with Celite R648 B biocatalysts . . . 124

5.3.5 BPA soluble reaction products formation kinetics: packed-bed reactor 126 5.4 Conclusion . . . 127

6 Dynamic measurement of bisphenol A removal based on oxygen consump- tion 129 6.1 Introduction . . . 129

6.2 Materials and Methods . . . 130

6.2.1 A non-invasive oxygen sensor . . . 130

6.2.2 Experimental configuration . . . 131

6.2.3 Evaluation of k

a , OCR and stoichiometric ratio R . . . 131

6.3 Results and discussion . . . 133

6.3.1 Validation on experimental data . . . 133

6.4 Conclusion . . . 136

7 Modeling bisphenol A degradation kinetics 139 7.1 Model development . . . 139

7.1.1 Classic Briggs-Haldane kinetics . . . 139

7.1.2 Pre-steady state hypothesis . . . 141

7.1.3 BPA insoluble reaction products kinetics prediction . . . 144

7.2 Validation on experimental data . . . 145

7.3 Conclusion . . . 147

8 General conclusions and perspectives 149 8.1 Development of new biocatalysts . . . 149

8.2 BPA degradation kinetics and reaction products formation kinetics . . . 150

8.3 Characterization of reaction products from BPA oxidation . . . 151

8.4 Methods of BPA and its reaction products monitoring . . . 152

8.5 Packed-bed reactor optimization and perspectives . . . 153

Bibliography 166

Appendices 167

A Immobilization yields Y and Y

calculation 168

B Residence Time Distribution: outlet conductivity σ

calculation 169

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CONTENTS

C Initial enzymes activities for Pleurotus ostreatus immobilization 170

D Polymerization kinetics: model resolution 171

E Mathematical definitions of M

, M

and M

172

F Influence of samples filtration 173

G Complementary DAD280 and EIC chromatograms from LC/MS analyses175 H Harnessing laccases for the synthesis of bisphenol A biopolymers 178

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