School of Engineering Department of Chemical Engineering CHARACTERIZATION OF S-OXIDIZING BIOMASS THROUGH RESPIROMETRIC TECHNIQUES UNDER ANOXIC AND AEROBIC CONDITIONS PhD Thesis Supervised by: Dr. David Gabriel Buguña & Dr. Xavier Gamisans Noguera MABEL MORA GARRIDO Bellaterra, October 2014 DAVID GABRIEL BUGUÑA, professor agregat del Departament d’Enginyeria Química de la Universitat Autònoma de Barcelona, i XAVIER GAMISANS NOGUERA, Catedràtic d’Escola Universitària, CERTIFIQUEM: Que l’enginyera química MARIA ISABEL MORA GARRIDO ha realitzat sota la nostra direcció el treball titulat: “CHARACTERIZATION OF S-OXIDIZING BIOMASS THROUGH RESPIROMETRIC TECHNIQUES UNDER ANOXIC AND AEROBIC CONDITIONS”, el qual es presenta en aquesta memòria i que constitueix la seva Tesi per a optar al Grau de Doctor per la Universitat Autònoma de Barcelona. I perquè en prengueu coneixement i consti als efectes oportuns, presentem a l’Escola d’Enginyeria de la Universitat Autònoma de Barcelona l’esmentada Tesi, signant el present certificat a Bellaterra, 30 d’octubre de 2014 David Gabriel Buguña Xavier Gamisans Noguera AAAA mmmmiiiissss áááánnnnggggeeeelllleeeessss ddddeeee llllaaaa gggguuuuaaaarrrrddddaaaa CONTENTS Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v Resum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vi Resumen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii List of Abbreviations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix Chapter 1 MOTIVATIONS AND THESIS OVERVIEW . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 Chapter 2 INTRODUCTION Desulfurization of biogas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 Chemistry of hydrogen sulfide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 Biological elemental sulfur . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 Biological sulfur metabolism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 Respirometric and titrimetric techniques to characterize SOB . . . . . . . . 22 Chapter 3 OBJECTIVES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31 Chapter 4 GENERAL MATERIALS AND METHODS Description of equipments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35 Analytical techniques . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44 Chapter 5 DEVELOPMENT OF AN ANOXIC RESPIROMETRIC TECHNIQUE TO CHARACTERIZE SO-NR MIXED CULTURES OBTAINED FROM DESULFURIZING BIOTRICKLING FILTERS Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53 Materials and methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54 SO-NR biomass cultivation Respirometric methodology development Experimental tests Results and discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59 Biomass growth yield calculation Thiobacillus denitrificans pure culture SO-NR mixed culture Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65 Chapter 6 COUPLING RESPIROMETRY AND TITRIMETRY TO INVESTIGATE THE KINETICS OF THIOSULFATE-DRIVEN AUTOTROPHIC DENITRIFICATION AND THE EFFECT OF NITRITE ACCLIMATION Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71 Materials and methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72 SO-NR biomass Respirometric and titrimetric tests Proton production rate Model development . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75 Stoichiometry of thiosulfate-driven autotrophic denitrification Modeling nitrite reduction Modeling two-step denitrification Parameters estimation Fisher information matrix Results and discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80 Stoichiometric coefficients Denitritation kinetics Calibration of two-step denitrification model Kinetic assessment of biomass nitrite acclimation Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92 Chapter 7 KINETIC AND STOICHIOMETRIC CHARACTERIZATION OF ANOXIC SULFIDE OXIDATION BY SO-NR MIXED CULTURES OBTAINED FROM ANOXIC BIOTRICKLING FILTERS Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 97 Materials and methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 97 S-oxidizing biomass Respirometric tests Carbon source and substrate stripping Kinetic model Results and discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102 Assessment of the SO-NR cultivation in a CSTR Characterization of sulfide and CO stripping 2 Kinetic analysis of sulfide and nitrite inhibitions Sulfide oxidation using nitrate as the electron acceptor Sulfide oxidation using nitrite as the electron acceptor Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 112 Chapter 8 PRELIMINARY STUDY ON LFS RESPIROMETRY APPLICATION TO CHARACTERIZE SULFIDE, ELEMENTAL SULFUR AND THIOSULFATE OXIDATION KINETICS AND STOICHIOMETRY UNDER AEROBIC CONDITIONS BY SOB Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 115 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117 Materials and methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119 SOB cultivation and biomass growth yield calculation Characterization of H S stripping and chemical oxidation 2 LFS respirometric tests Model development . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121 Sulfide biological oxidation Thiosulfate production and biodegradation Elemental sulfur biological oxidation Oxygen uptake rate Parameters estimation Results and discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125 Assessment of SOB cultivation in a CSTR H S stripping characterization 2 Chemical oxidation of sulfide Biological sulfide oxidation mechanisms Stoichiometry of the process Kinetic model calibration Validation of the kinetic model Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137 Chapter 9 STUDY ON THE APPLICATION OF HETEROGENEOUS RESPIROMETRY TO CHARACTERIZE H S OXIDIZING BIOFILM FROM DESULFURIZING 2 BIOTRICKLING FILTERS Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 143 Materials and methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 144 Immobilized S-oxidizing biomass cultivation Experimental approach of the heterogeneous respirometry Experimental determinations in the packed bed Mathematical model development . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 146 Mass balance for the gas phase Mass balance for the liquid phase Mass balance for the biofilm Microbial kinetic model Stoichiometry of H S oxidation 2 Model parameters estimation Results and discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 152 Biomass cultivation Abiotic tests Biotic tests: estimation of kinetic parameters Assessment of the rate controlling step Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161 Chapter 10 GENERAL CONCLUSIONS AND FUTURE WORK . . . . . . . . . . . . . . . . . . . . . . 165 Chapter 11 REFERENCES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171
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