Grantee Research Project Results

Final Report: Inhibition, Inactivation and Recovery: A Universal Model for Aerobic Cometabolic Degradation of Aliphatic Compounds

EPA Grant Number: R825689C013
Subproject: this is subproject number 013 , established and managed by the Center Director under grant R825689
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
Center: National Research Program on Design-Based/Model-Assisted Survey Methodology for Aquatic Resources
Center Director: Stevens, Don L.
Title: Inhibition, Inactivation and Recovery: A Universal Model for Aerobic Cometabolic Degradation of Aliphatic Compounds
Investigators: Arp, Daniel J. , Williamson, Kenneth J. , Hyman, Michael R.
Institution: Oregon State University
EPA Project Officer: Hahn, Intaek
Project Period: January 1, 1996 through January 1, 1997
Project Amount: Refer to main center abstract for funding details.
RFA: Hazardous Substance Research Centers - HSRC (1989) RFA Text |  Recipients Lists
Research Category: Hazardous Substance Research Centers , Land and Waste Management

Objective:

The aims of this project are two-fold. First, the further development and corroboration will be done of a mathematical model previously developed which describes the kinetics of aerobic cometabolism of trichloroethylene (TCE). This model was originally developed for the soil nitrifying bacterium, Nitrosomonas europaea and describes TCE cometabolism in terms of an inhibition, an inactivation and a recovery process. The present study aims to extend these earlier studies to other organisms including toluene- and methane-oxidizing species. The second aim is to further investigate the physiological responses of N. europaea to the toxicity associated with the cometabolism of trichloroethylene.

Summary/Accomplishments (Outputs/Outcomes):

In situ bioremediation schemes have a considerable need for mathematical models that can accurately describe the kinetics of pollutant degradation. In particular, accurate models can provide important information for process design and for the interpretation of degradation studies. At a more fundamental level, there is also a considerable need to understand the molecular basis for the toxicity of various pollutants towards degradative organisms. In addition, it is important to understand the responses of the affected organisms to these toxic effects because these responses in turn affect the kinetics and modeling of the degradation process. In combination these large and small scale approaches should provide a comprehensive description of the kinetics and molecular consequences of aerobic cometabolism of chlorinated solvents.

The current approach for extending the modeling efforts is based on the quasi-steady-state reactor system which was previously developed for studies with N. europaea. In summary, a 1.7 gas-tight reactor vessel containing 0.5 L of reaction mixture is rapidly stirred using a magnetic stir bar. Primary substrates (e.g. ammonium) are supplied at a constant rate. Once steady state rates of activity have been established, an alternative pollutant substrate is added to the reactor and the kinetics of both the primary and secondary substrate degradation are monitored over a period of several hours. In addition, assays of residual enzyme activities are determined for small (< 2ml) samples removed from the reactor at regular intervals. Experiments investigating the mechanism and response of N. europaea to the toxicity associated with TCE and chlorinated solvent degradation are conducted on a smaller scale and over shorter times. The effects of these exposures are correlated with the losses of various enzyme activities and the response of the cells is subsequently monitored using radiolabeled 14CO2 to monitor de novo protein synthesis.

In previous studies, a methodology for conducting reactor-based studies with N. europaea was developed, and the mathematical model describing TCE cometabolism by this organism was formulated. Experiments were conducted with Burkholderia cepacia G4 to examine whether the model accurately describes the cometabolism of TCE by other organisms. The model does not accurately describe cometabolism in B. cepacia G4. Factors such as growth and substrate inhibition need to be further examined.

Short term, small scale assays examining inhibition, inactivation, and recovery associated with TCE cometabolism by the toluene oxidizer B. cepacia G4 have been initiated. These results reveal additional complexity in whole cell kinetics of toluene and TCE oxidation by B. cepacia G4 and further support the idea that mathematical models employing competitive inhibition may not adequately describe the kinetics of cometabolism. The effects of alkynes on toluene-2-monooxygenase (TOM) activity are under investigation. Although acetylene is a weak inactivator, longer chain alkynes are potent and specific inactivators of TOM.

The mechanism of TCE toxicity for N. europaea has been further investigated using short-term incubations. TCE cometabolism has significant effects on both AMO and HAO activities. Inhibitor treatments which specifically inactivate either AMO or HAO were used to compare effects seen in TCE cometabolism and several other chlorinated hydrocarbons. The results of TCE and chloroform cometabolism show that HAO is the probable target at low concentrations. In contrast, AMO is inactivated in cis-1,2-dichloroethylene experiments. These results lead us to conclude the site of action is not directly related to these enzymes. One hypothesis is that TCE cometabolism exerts a significant effect on the electron transport in N. europaea cells. Project completed.

Journal Articles on this Report : 1 Displayed | Download in RIS Format

Publications Views

Other subproject views:	All 2 publications	1 publications in selected types	All 1 journal articles
Other center views:	All 385 publications	251 publications in selected types	All 194 journal articles

Publications

Type	Citation	Sub Project	Document Sources
Journal Article	Ely RL, Williamson KJ, Hyman MR, Arp DJ. Cometabolism of chlorinated solvents by nitrifying bacteria: kinetics, substrate interactions, toxicity effects, and bacterial response. Biotechnology and Bioengineering 1997;54(6):520-534.	R825689C013 (Final)	Abstract from PubMed Abstract: Wiley-Abstract Exit

Supplemental Keywords:

Groundwater, chlorinated organics, biodegradation, trichloroethylene, degradation models, aerobic cometabolism., RFA, Scientific Discipline, Toxics, Waste, National Recommended Water Quality, Environmental Chemistry, Remediation, Chemistry, Environmental Microbiology, Hazardous Waste, Bioremediation, Engineering, Environmental Engineering, 33/50, Groundwater remediation, Hazardous, soil nitrifying bacterium, degradation, chlorinated solvent, in situ remediation, degradation models, Tetrachloroethylene, biodegradation, cometabolism, recovery, aerobic cometabolism, kinetic studies, chlorinated organic compounds, in situ bioremediation, in-situ bioremediation, mathematical models, models, chlorinated organics, bacterial degradation, chlorinated solvents, degrade trichloroethylene, groundwater, TCE

Main Center Abstract and Reports:

R825689    National Research Program on Design-Based/Model-Assisted Survey Methodology for Aquatic Resources

Subprojects under this Center: (EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
R825689C001 Interactions between Electron Acceptors in the Treatment of Wastewaters Containing Sulfate, Chlorophenols and Acetate
R825689C002 Enhancing Biodegradation with Sorption and Alternating Aerobic/Anaerobic Environments
R825689C003 Development and Verification of a Numerical Model to Predict the Fate and Transport of Chlorinated Phenols in Groundwater
R825689C004 Redox Transformations of Inorganic Pollutants: Coupling to the Biogeochemical Matrix
R825689C005 Hexavalent Chromium Sorption and Desorption in Natural Soils and Subsoils
R825689C006 Biotransformation of Ordnance Wastes Using Unique Consortia of Anaerobic Bacteria
R825689C007 The Effect of Environmental Conditions on Reductive Dechlorination Rates
R825689C008 Lead Sorption, Transport, and Remediation in Natural Soils and Subsoils
R825689C009 Degradation of Chlorinated Aliphatic Compounds by Nitrifying Bacteria
R825689C010 Remediation of Contaminated Soil from the Baldock Station Maintenance Facility
R825689C011 The Effect of Apparent EH, Compound Structure, and Electron Donor on Anaerobic Biotransformation of Trinitrotoluene and its Metabolites
R825689C012 The "Bubble Wall": A Passive In Situ System for Treatment and/or Containment of Contaminated Groundwater
R825689C013 Inhibition, Inactivation and Recovery: A Universal Model for Aerobic Cometabolic Degradation of Aliphatic Compounds
R825689C014 Development, Characterization, and Performance Evaluation of Ferrous-Ferric Oxide Adsorbents for Metal Removal from Contaminated Groundwater
R825689C015 Redox Transformations of Organic and Inorganic Contaminants in the Subsurface Environment
R825689C016 Demonstration of a Permeable Barrier Technology for the Bioremediation of Ground Water Contaminated with Waste Mixtures
R825689C017 Development of a Vitamin B12-Amended Bioremediation Process for the Reductive Dechlorination of Chlorobiphenyls at all Chlorine Positions
R825689C018 An Investigation of the Chlorinated Hydrocarbon Substrate Range of the Filamentous Fungus, Graphium sp.
R825689C019 Aerobic Cometabolism of Chloroform, 1,1,1-trichloroethane, 1,1-dichloroethylene, and Other Chlorinated Aliphatic Hydrocarbons by Microbes Grown on Butane and Propane
R825689C020 Cytochrome P-450: An Emerging Catalyst for the Cometabolism of Chlorinated Aliphatic Hydrocarbons and Methyl tert-butyl Ether?
R825689C021 In-Situ Measurement of TCE Degradation Using a Single-Well "Push-Pull" Test
R825689C022 Development and Characterization of Redox Sensors for Environmental Monitoring
R825689C023 Assessing Metal Speciation in the Subsurface Environment
R825689C024 Simultaneous Removal of the Adsorbable and Electroactive Metals from Contaminated Soils and Groundwater
R825689C025 Multisolute Sorption and Transport Model for Copper, Chromium, and Arsenic Sorption on an Iron-Coated Sand, Synthetic Groundwater System
R825689C026 Development of Alkoxysilanes as Slow Release Substrates for the Anaerobic/Aerobic Transformation of Chlorinated Solvents
R825689C027 Aerobic Cometabolism of Chlorinated Aliphatic Hydrocarbons by Toluene-Oxidizing Bacteria
R825689C028 Development and Characterization of Sensors and Field Instrumentation for Monitoring of Environmental Redox Conditions
R825689C029 Aerobic Cometabolism of Methyl tert-butyl Ether by Microorganisms Grown on Aliphatic Hydrocarbons
R825689C030 Biotransformation of Lead and Chromate by Bacteria
R825689C031 Magnetic Resonance Studies of Heavy Metals in Clays, Zeolites and Ceramics
R825689C032 Probing the Redox Properties of Environmental Systems: Natural Phenolic Materials
R825689C033 Reductive Dehalogenation at Carbon and Derivatized Carbon Electrodes
R825689C034 Detection of Microorganisms Capable of Anaerobic Degradation of Hazardous Substances in Natural Environments
R825689C035 Treatment of Complex Mixtures
R825689C036 Oxidation of Chlorinated Solvents by Methanotrophs
R825689C037 Detection and Assessment of Subsurface Contamination
R825689C038 Design of Reliable and Cost-Effective Mitigation Schemes
R825689C039 Gaseous Stripping of Nonaqueous Phase Liquids from the Vadose Zone
R825689C040 Anaerobic Microbial Transformation of Homocyclic and Heterocyclic Polynuclear Aromatic Hydrocarbons
R825689C041 Effects of Sorption on Biodegradation of Halogenated Organics
R825689C042 Trace Metal Removal Processes
R825689C043 FASTCHEM Applications and Sensitivity Analysis
R825689C044 Long-term Chemical Transformation of 1,1,1-Trichloroethane (TCA) and Freon 113 under Aquifer Conditions
R825689C045 In-Situ Anaerobic Biological Treatment of Aromatics in Groundwater
R825689C046 Use of Starvation and Stress Promoters for Biodegradation of Hazardous Wastes
R825689C047 Determining and Modeling Diffusion-Limited Sorption and Desorption Rates of Organic Contaminants in Heterogeneous Soils
R825689C048 Dispersion Modeling of Volatile Organic Emissions from Ground-Level Treatment Systems
R825689C049 Subsurface Mixing of Nutrients and Groundwater for in-Situ Bioremediation
R825689C050 Test-Bed Evaluation of In-Situ Bioremediation of Chlorinated Aliphatic Compounds by Toluene Oxygenase Microorganisms
R825689C051 Demonstration of in-Situ Bioremediation of Chlorinated Aliphatics by Methanotrophs at St. Joseph
R825689C052 Aquifer Remediation Design in the Presence of Kinetic Limitations
R825689C053 Determination of Macroscopic Transport Parameters for Biologically Reacting Solutes in Aquifers
R825689C054 Transformation of Chlorinated Hydrocarbons by Reduced Metallocoenzymes--Kinetic Model Development and Applications to Environmental Systems
R825689C055 Microbial Degradation of Toluene Under Sulfate-Reducing Conditions--The Role of Iron
R825689C056 Transformation of TCE by Methanotrophic Biofilms
R825689C057 Heavy Metals in Ceramic Matrix: Heavy Metals/Clay Interactions in Ceramic Processing
R825689C058 Radon-222 Method for Locating and Quantifying Contamination by Residual Non-Aqueous Phase Liquids in the Subsurface
R825689C059 Process Submodel Formulation and Parameter Estimation for Simulation of Bioremediation
R825689C060 Enhancement of Biodegradation through the Use of Substituted Porphyrins to Treat Groundwater Contaminated with Halogenated Aliphatics
R825689C061 Field Test of In-Situ Vapor Stripping for Removal of VOCS from Groundwater
R825689C062 System Design for Enhanced In-Situ Biotransformation of Carbon Tetrachloride: Application to DOE's Arid Site Integrated Demonstration
R825689C063 Modeling Strategies for Optimizing In-Situ Bioremediation
R825689C064 Anaerobic Treatment of Chlorinated Solvent Contaminated Groundwater
R825689C065 In Situ Treatment of Chlorinated Solvents
R825689C066 Moffett Field In-Situ Bioremediation Study in Support of Full Scale Application
R825689C067 Full-Scale Evaluation of In Situ Bioremediation of Chlorinated Solvent Groundwater Contamination
R825689C068 Upscaling Pore-Scale Hydrodynamics and the Transport of Reactive Solutes
R825689C069 Pathways of Anaerobic Toluene Metabolism by a Sulfate-Reducing Bacterium, Strain PRTOL1
R825689C070 Anaerobic Ethylbenzene Oxidation in Denitrifying Strain EB1
R825689C071 Molecular Approaches to Optimize Starvation Promoter Dricen TCE Bioremediation in Pseudomonas
R825689C072 Modeling VOC Emissions from Hazardous Waste Sites
R825689C073 Reductive Transformation of Chlorinated Hydrocarbons by Reduced Ethenes Catalyzed by Vitamin B12 - Mechanistic and Kinetic Studies
R825689C074 Evaluation of Strategies for Full Scale Bioremediation of the Seal Beach Site Using Anaerobic Microbial Processes
R825689C075 Trace Element Adsorption in Porous Particle Packed Beds
R825689C076 Hydrologic and Biological Factors Affecting Aquifer Clogging During In-Situ Bioremediation
R825689C077 Full-Scale Evaluation of an Apparatus for Down-well Oxygen Transfer to Implement In situ Bioremediation at Edwards AFB
R825689C078 Field Testing of Palladium-Catalyzed Hydrodehalogenation for Chlorinated Hydrocarbon Removal from Groundwater
R825689C079 Physics of Dissolution of Nonaqueous Phase Liquids: Pore Networks and Field Simulations
R825689C080 Three-Phase Flow in Fractured Media
R825689C081 Effects of Redox Zones on the Fate and Transport of Contaminants in the Saturated Subsurface; Characterization and Simulation
R825689C082 Biochemical Mechanisms of PCE Dehalogenation by Strain MS-1, and its Potential for In-situ Bioaugmentation
R825689C083 A Large Scale Model for Anaerobic Bioremediation at the Seal Beach Site
R825689C084 Mechanisms, Chemistry, and Kinetics of Anaerobic Degradation of cDCE and Vinyl Chloride
R825689C085 Bioenhanced In-Well Vapor Stripping to Treat Trichloroethylene (TCE)
R825689C086 Effect of Chemical Structure on the Biodegradability of Halogenated Hydro-carbons
R825689C087 Trace Element Adsorption in Porous Particle Packed Beds
R825689C088 Arsenic Removal in High Capacity Porous Alumina Packed-Bed Reactors
R825689C089 Measurement of Interfacial Areas and Mass Transfer Coefficients Between Residual PCE and Water During Surfactant Enhanced Aquifer Remediation
R825689C090 Proof of Gene Expression During Bioaugmentation
R825689C091 Experimental and Mathematical Study of Biomass Growth in Pore Networks and its Consequences in Bioremediation
R825689C092 Gene probes for detecting anaerobic alkylbenzene-degrading bacteria
R825689C093 Investigation of Palladium Catalyzed Hydrodehalogenation for the Removal of Chlorinated Groundwater Contaminants: Surface Chemistry of Catalyst Deactivation and Regeneration
R825689C094 Aerobic Methanotrophic Transformation of Biphenyl, Monochlorobiphenyls, and Dichlorobiphenyls