BOOK
Immobilised Biocatalysts for Bioremediation of Groundwater and Wastewater
Rita Hochstrat | Thomas Wintgens | Philippe Corvini
(2015)
Additional Information
Book Details
Abstract
The European project MINOTAURUS explored innovative bio-processes to eliminate emerging and classic organic pollutants. These bio-processes are all based on the concept of immobilization of biocatalysts (microorganisms and enzymes) and encompass bioaugmentation, enzyme technology, rhizoremediation with halophytes, and a bioelectrochemical remediation process. The immobilization-based technologies are applied as engineered ex situ treatment systems as well as natural systems in situ for the bioremediation of groundwater, wastewater and soil. The selection and application of tailored physico-chemical, molecularbiological and ecotoxicological monitoring tools combined with a rational understanding of engineering, enzymology and microbial physiology is a pertinent approach to open the black-box of the selected technologies. Reliable process monitoring constitutes the basis for developing and refining biodegradation kinetics models, which in turn improve the predictability of performances to be achieved with technologies.Â
Immobilised Biocatalysts for Bioremediation of Groundwater and Wastewater delivers insight into the concepts and performance of a series of remediation approaches. A key strength of this book is to deliver results from lab-scale through to piloting at different European reference sites. It further suggests frameworks for structuring and making evidence-based decisions for the most appropriate bioremediation measures.
Table of Contents
| Section Title | Page | Action | Price |
|---|---|---|---|
| Cover | Cover | ||
| Contents | v | ||
| List of figures | ix | ||
| List of tables | xiii | ||
| List of contributors | xv | ||
| Preface | xvii | ||
| Acknowledgement | xix | ||
| Abbreviations | xxi | ||
| Chapter 1: Introduction | 1 | ||
| 1.1 POLLUTANTS IN THE AQUATIC ENVIRONMENT | 1 | ||
| 1.1.1 Types, occurrence and fate | 1 | ||
| 1.1.2 Regulatory frameworks | 6 | ||
| 1.1.2.1 EU level legislation | 6 | ||
| 1.1.2.2 National legislation | 7 | ||
| 1.2 ENVIRONMENTAL BIOTECHNOLOGY OPTIONS | 7 | ||
| 1.2.1 Challenges for the implementation of bioaugmentation | 7 | ||
| 1.2.2 Challenges for the use of enzymes | 7 | ||
| 1.2.3 Immobilization of whole cells and enzymes as a solution of choice to circumvent bioremediation difficulties | 8 | ||
| 1.3 RECENT RESEARCH – THE MINOTAURUS PROJECT | 8 | ||
| 1.3.1 Scope and ambition | 8 | ||
| 1.3.2 Approach | 9 | ||
| 1.4 ABOUT THIS BOOK | 10 | ||
| 1.5 REFERENCES | 12 | ||
| Chapter 2: Analytical and monitoring methods | 15 | ||
| 2.1 CHEMICAL METHODS FOR THE ANALYSIS OF TARGET POLLUTANTS | 16 | ||
| 2.1.1 Endocrine disrupting compounds and pharmaceuticals | 16 | ||
| 2.1.1.1 Liquid chromatography for the analysis of polar pharmaceuticals | 17 | ||
| SMX CBZ and DF measurement with HPLC-MS | 17 | ||
| 2.1.1.2 Analysis of BPA with GC-MS | 18 | ||
| 2.1.1.3 Simulateneous measurement of BPA, CBZ, DF, EE2, NP, SMX, and TCS | 18 | ||
| UHPLC method | 18 | ||
| 2.1.2 Methyl tert-butyl ether, tert-butyl alcohol and chlorinated aliphatic hydrocarbons | 19 | ||
| 2.2 ISOTOPIC METHODS | 19 | ||
| 2.2.1 14C-Radioanalytics | 19 | ||
| 2.2.1.1 Background and potential | 19 | ||
| 2.2.1.2 Application in the MINOTAURUS project | 20 | ||
| Example of radiomonitoring study | 20 | ||
| Standardized radioactive assay for degradation capacity of microbes | 21 | ||
| 2.2.2 Compound specific stable isotope analysis (CSIA) | 22 | ||
| 2.2.2.1 Exemplified CSIA application in a technical processes | 23 | ||
| 2.3 BIOCATALYST MONITORING | 24 | ||
| 2.3.1 Monitoring tools for microorganisms | 24 | ||
| 2.3.1.1 Fluorescence in situ hybridization (FISH) | 25 | ||
| 2.3.1.2 Quantitative polymerases chain reaction (qPCR) | 26 | ||
| Principle of qPCR assay based on SYBR Green I reaction chemistry | 26 | ||
| Quantification of gene copy numbers | 26 | ||
| Optimization of qPCR assay | 28 | ||
| qPCR assay to quantify hqdB gene of Sphingomonas sp. strain TTNP3 in bioaugmented MBR | 28 | ||
| qPCR assay targeting 16S rRNA gene of Microbacterium sp. BR1 | 30 | ||
| 2.3.1.3 Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) | 30 | ||
| 2.3.1.4 Next generation sequencing | 31 | ||
| 2.3.1.5 Stable isotope probing (SIP) | 32 | ||
| 2.3.1.6 BACTRAPs | 33 | ||
| Concept and working principle | 33 | ||
| Field application in benzene contaminated site | 34 | ||
| Results of the experiments on microbial benzene transformation in constructed wetlands in Leuna | 34 | ||
| Assessing the suitability of the BACTRAP approach for identifying BPA-degrading microbes | 36 | ||
| Mineralization of BPA in a MBR-Reactor and by Sphingomonas sp. strain TTNP3 | 36 | ||
| Conclusions and recommendation for the application of BACTRAPs in remediation processes | 36 | ||
| 2.3.2 Monitoring tools for enzymes | 37 | ||
| 2.3.2.1 Colorimetric assays for measuring laccase activity | 38 | ||
| ABTS oxidation assay for routine laccase activity determination | 38 | ||
| 2,6 Dimethoxyphenol assay (substrate surrogate) | 38 | ||
| 2.3.2.2 Determining oxygen consumption rate (OCR) | 39 | ||
| Microplate-based flux analyzer | 39 | ||
| Clark electrode | 39 | ||
| Oxygen optical sensor | 40 | ||
| 2.3.2.3 Determining enzyme activity via co-factor oxidation in the UV-range | 41 | ||
| Monooxygenase assay | 41 | ||
| Fungal peroxygenase (AaP) assay | 42 | ||
| 2.3.2.4 Assessment of enzyme kinetics with target pollutants using chemical analysis | 42 | ||
| 2.4 ECOTOXICITY MONITORING | 43 | ||
| 2.4.1 Batteries of ecotoxicity tests | 43 | ||
| 2.4.2 Ecotoxicity tests used within the MINOTAURUS project | 44 | ||
| 2.5 REFERENCES | 44 | ||
| Chapter 3: Immobilization techniques for biocatalysts | 49 | ||
| 3.1 INTRODUCTION | 49 | ||
| 3.2 IMMOBILIZATION OF BIOMASS | 50 | ||
| 3.2.1 Bioaugmented membrane bioreactor (MBR) | 50 | ||
| 3.2.1.1 Immobilization on carrier material | 50 | ||
| Sphingomonas sp. strain TTNP3 | 50 | ||
| Phoma sp. strain UHH 5-1-03 | 51 | ||
| 3.2.1.2 Encapsulation in alginate beads | 52 | ||
| Procedure | 52 | ||
| Degradation performance in application relevant conditions | 53 | ||
| 3.2.2 Bioaugmented packed bed reactor (PBR) | 54 | ||
| 3.2.2.1 Biomass immobilization for the development of packed bed reactors treating chlorinated aliphatic hydrocarbons (CAH) | 54 | ||
| 3.2.3 Microorganisms on electrically conductive carriers | 57 | ||
| 3.2.3.1 Tailored electrically-conductive carrier materials | 59 | ||
| 3.3 IMMOBILIZATION OF ENZYMES | 59 | ||
| 3.3.1 Bio-inspired titanification | 59 | ||
| 3.3.1.1 Principle | 59 | ||
| 3.3.1.2 Biocatalyst performance | 61 | ||
| 3.3.2 Enzyme conjugated nanoparticles | 62 | ||
| 3.3.2.1 Principle | 62 | ||
| 3.3.2.2 Biocatalyst performance | 63 | ||
| Immobilization effectiveness of single laccases | 63 | ||
| Stability of single laccase conjugates | 64 | ||
| Degradation of target compounds | 64 | ||
| Immobilization of combinations of laccase on the same carrier | 65 | ||
| Properties of free and immobilized laccases regarding target transformation | 65 | ||
| Oxidation of the target micropollutants | 66 | ||
| Stability | 67 | ||
| 3.4 REFERENCES | 67 | ||
| Chapter 4: Bioaugmented membrane bioreactor technology | 71 | ||
| 4.1 STATE OF THE ART | 71 | ||
| 4.2 PROCESS DESCRIPTION | 71 | ||
| 4.2.1 Pilot-scale set-up | 72 | ||
| 4.2.2 Bioaugmentation | 73 | ||
| 4.2.3 Sampling and analytical methods | 73 | ||
| 4.3 BASIC DESIGN PRINCIPLE | 73 | ||
| 4.4 OPERATIONAL MODES, EXPERIENCES, RESULTS | 74 | ||
| 4.4.1 General operational experience, removal of macropollutants | 74 | ||
| 4.4.2 Removal of BPA | 75 | ||
| 4.4.3 Biomass characterization by real time PCR | 76 | ||
| 4.5 UNRESOLVED ISSUES | 76 | ||
| 4.6 POTENTIAL APPLICATION SCENARIOS | 77 | ||
| 4.7 REFERENCES | 77 | ||
| Chapter 5: Enzyme reactors | 79 | ||
| 5.1 STATE OF THE ART | 79 | ||
| 5.2 ENZYMATIC MEMBRANE REACTOR | 81 | ||
| 5.2.1 Laboratory application of laccase-conjugated nanoparticles for micropollutants removal | 81 | ||
| 5.2.2 Application of GTL-fsNPs in advanced wastewater treatment | 82 | ||
| 5.2.2.1 Reactor concept | 82 | ||
| 5.2.2.2 Laboratory bench-scale reactor | 82 | ||
| 5.2.2.3 Field pilot-scale plant | 83 | ||
| 5.2.3 Operational modes, results, experience | 84 | ||
| 5.2.3.1 Laboratory bench-scale experiments | 84 | ||
| 5.2.3.2 Results of pilot-scale experiments | 85 | ||
| Pilot trial 1 | 85 | ||
| Pilot trial 2 | 85 | ||
| Pilot trial 3 | 86 | ||
| Pilot trial 1 | 86 | ||
| Removal of BPA during pilot trial 1 | 86 | ||
| Enzyme activity and stability of GTL-fsNPs | 88 | ||
| Pilot trials 2 and 3 | 88 | ||
| Removal of target micropollutants during pilot trials 2 and 3 | 88 | ||
| Enzyme activity and stability in pilot trial 2 and 3 | 89 | ||
| Influence of suspended solids | 89 | ||
| Influence of GTL-fsNPs on the membrane filtration | 90 | ||
| 5.2.4 Unresolved issues | 90 | ||
| 5.2.5 Potential application scenarios | 91 | ||
| 5.3 MAGNETIC RETENTION REACTOR | 91 | ||
| 5.3.1 Reactor design and process description | 91 | ||
| 5.3.2 Preliminary micropollutant removal tests in batch | 92 | ||
| 5.3.3 Continuous operation of the reactor with magnetic retention | 93 | ||
| 5.4 REFERENCES | 95 | ||
| Chapter 6: Rhizodegradation in constructed wetlands | 97 | ||
| 6.1 STATE OF THE ART AND PROCESS DESCRIPTION | 97 | ||
| 6.2 BASIC DESIGN PRINCIPLES | 99 | ||
| 6.2.1 Vegetation | 99 | ||
| 6.2.2 Design considerations | 99 | ||
| 6.2.2.1 BOD removal | 99 | ||
| 6.2.2.2 Total suspended solids (TSS) removal | 99 | ||
| 6.2.2.3 Nitrogen removal | 99 | ||
| 6.2.2.4 Phosphorus removal | 100 | ||
| 6.2.2.5 Pathogens removal | 100 | ||
| 6.2.3 Hydrological balance of CWs | 100 | ||
| 6.2.3.1 Evapotranspiration | 100 | ||
| 6.2.4 General design parameters | 100 | ||
| 6.2.5 Construction details | 101 | ||
| 6.2.6 General design for contaminants removal | 101 | ||
| 6.2.7 Hydraulic design of horizontal subsurface flow systems | 102 | ||
| 6.3 EXPECTED PERFORMANCE AND FIELD EXPERIENCE | 102 | ||
| 6.3.1 The MINOTAURUS project experience | 102 | ||
| 6.3.2 Contribution of endophytic bacteria in rhizodegradation | 103 | ||
| 6.4 UNRESOLVED ISSUES | 103 | ||
| 6.5 POTENTIAL APPLICATION SCENARIOS | 104 | ||
| 6.6 REFERENCES | 104 | ||
| Chapter 7: Packed bed reactors (PBR) to treat chlorinated aliphatic hydrocarbons via aerobic cometabolism as pump & treat technology | 107 | ||
| 7.1 STATE OF THE ART | 107 | ||
| 7.2 PROCESS DESCRIPITION | 108 | ||
| 7.2.1 Aerobic cometabolic biodegradation of chlorinated aliphatic hydrocarbons | 108 | ||
| 7.2.2 Pulsed feed of oxygen and growth substrate | 109 | ||
| 7.3 BASIC DESIGN PRINCIPLE | 110 | ||
| 7.3.1 Selection of the growth substrate and development of a suitable suspended-cell microbial consortium | 110 | ||
| 7.3.2 Selection of the biofilm carrier | 110 | ||
| 7.3.3 Kinetic study to characterize the target CAHs biodegradation by the attached-cell consortium | 110 | ||
| 7.3.4 Fluid-dynamic characterization of a packing of the selected biofilm carrier | 111 | ||
| 7.3.5 Analysis of the process robustness and reliability | 111 | ||
| 7.3.6 Sizing of the PBR and preliminary design of the schedule of pulsed oxygen/substrate supply | 112 | ||
| 7.4 OPERATIONAL MODES, EXPERIENCES, RESULTS | 113 | ||
| 7.4.1 Growth substrate selection and consortium characterization | 113 | ||
| 7.4.2 Biofilm carrier selection | 114 | ||
| 7.4.3 Kinetic study of TCE biodegradation by the selected attached-cell consortium | 114 | ||
| 7.4.4 Fluid-dynamic characterization of the selected biofilm carrier | 116 | ||
| 7.4.5 Analysis of the process robustness and reliability | 116 | ||
| 7.4.6 Application of the PBR sizing procedure and operation of a 31-L pilot-scale PBR | 118 | ||
| 7.5 UNRESOLVED ISSUES (FROM PILOT TO FIELD) | 121 | ||
| Inclusion in the kinetic model of the terms accounting for the production and consumption of reducing power (NADH) | 121 | ||
| Attainment of high growth substrate concentrations at the PBR inlet | 121 | ||
| Possible stripping of volatile CAHs, as a result of the solubilization of gaseous substrate at the PBR inlet | 121 | ||
| Attaining of very high CAH degradation yields, so as to comply with the limits for the remediation of CAH-contaminated sites | 121 | ||
| 7.6 POTENTIAL APPLICATION SCENARIOS | 122 | ||
| 7.7 REFERENCES | 122 | ||
| Chapter 8: Packed bed reactors (pump & treat technologies) to treat MTBE/TBA contaminated groundwater | 125 | ||
| 8.1 STATE OF THE ART | 125 | ||
| 8.2 PROCESS DESCRIPITION | 125 | ||
| 8.3 BASIC DESIGN PRINCIPLE | 126 | ||
| 8.3.1 Working area of the M-consortium | 126 | ||
| 8.3.2 Retention of biomass in the bioreactor | 126 | ||
| 8.3.3 Reliability of the inoculated bioreactor | 127 | ||
| 8.3.4 Robustness of the inoculated bioreactor | 127 | ||
| 8.4 OPERATIONAL MODES, EXPERIENCES, RESULTS | 128 | ||
| 8.4.1 Evaluation of carrier materials | 128 | ||
| 8.4.1.1 Compatibility test | 128 | ||
| 8.4.1.2 Retention of biomass | 129 | ||
| 8.4.1.3 Fluidisation | 130 | ||
| 8.4.1.4 Conclusions | 131 | ||
| 8.4.2 Degradation performance in bench-scale 7 L bioreactor systems | 131 | ||
| 8.4.2.1 Recirculation mode with spiked influent | 131 | ||
| 8.4.2.2 Continuous mode with real groundwater | 131 | ||
| 8.4.2.3 Quantification and characterisation of biomass | 132 | ||
| 8.4.3 Pilot-scale bioreactor tests | 133 | ||
| 8.4.3.1 Test approach | 133 | ||
| 8.4.3.2 Test results | 134 | ||
| 8.4.3.3 Conclusions | 135 | ||
| 8.5 UNRESOLVED ISSUES (FROM PILOT TO FIELD) | 135 | ||
| 8.6 POTENTIAL APPLICATION SCENARIOS | 135 | ||
| 8.7 REFERENCES | 136 | ||
| Chapter 9: Bioelectrochemical reactor (in situ remediation) | 137 | ||
| 9.1 STATE OF THE ART | 137 | ||
| 9.2 PROCESS DESCRIPTION | 137 | ||
| 9.3 BASIC DESIGN PRINCIPLE | 139 | ||
| 9.4 OPERATIONAL MODES, EXPERIENCES, RESULTS | 139 | ||
| 9.4.1 Cathodic reductive dechlorination of TCE | 139 | ||
| 9.4.1.1 Influence of the cathode potential on the rate and yield of the reductive dechlorination process | 139 | ||
| 9.4.1.2 Microbiological characterization of the biocathode | 142 | ||
| 9.4.2 Anodic removal of chlorinated intermediates of cathodic dechlorination | 144 | ||
| 9.4.2.1 Assessment of the potential for anaerobic electrochemical oxidation of cis-DCE and ethene | 144 | ||
| 9.4.2.2 Microbial oxidation of cis-DCE and ethene sustained by electrolytic oxygen generation | 145 | ||
| 9.4.3 Overall performance of the sequential cathodic/anodic bioelectrochemical remediation process | 148 | ||
| 9.5 UNRESOLVED ISSUES | 149 | ||
| 9.6 POTENTIAL APPLICATION SCENARIOS | 149 | ||
| 9.7 REFERENCES | 149 |