Program Element 1: Biotransformation and Biodegradation

Fundamental research in microbiology to elucidate the mechanisms of biotransformation and biodegradation of complex contaminant mixtures.

PROGRAM OBJECTIVE


To understand the mechanisms and pathways for biotransformation and biodegradation of contaminant mixtures in order to improve the performance of bioremediation in the field.

BACKGROUND



The current understanding of biotransformation and biodegradation pathways and mechanisms in the field is incomplete.


The goal of all bioremediation efforts is to reduce the potential toxicity of chemical contaminants in the field by using microorganisms, plants, and animals to transform, degrade, or immobilize toxicants. There is already a significant base of knowledge about many pathways for organic degradation, and several important contaminant degradation mechanisms are presently under detailed investigation. Fundamental knowledge of biodegradation and biotransformation mechanisms has already contributed significantly to improving the performance of bioremediation in the field. Bioremediation strategies employing co-metabolism, anaerobic biotransformations of highly chlorinated solvents, and alternate electron acceptors which are in use today resulted in large part from fundamental investigations of the mechanisms used by microorganisms to transform contaminants.

Despite the successful contributions of existing knowledge about biodegradation and biotransformation mechanisms, there is still much that research can contribute. At present, the understanding of biotransformation and biodegradation pathways and mechanisms in the field is incomplete. Although the degradation of many organics and the biotransformation of some inorganic compounds in laboratory cultures have been well described, it is unclear how this information relates to bioremediation processes under field conditions. In addition, research is needed to understand recently discovered biotransformation processes such as metal biotransformations and biosequestration, coupled aerobic and anaerobic processes, co-metabolism, biotransformations in the presence of alternative electron donors/acceptors, and biotransformations catalyzed by consortia (Ehrlich, 1990; Francis, 1990; Lovley, 1993; Macaskie, 1991).

In both the short and long term, a sound understanding of the fundamental mechanisms of biotransformation and detoxification will lead to improved prediction, control, and assessment of bioremediation performance, facilitate the selection and prioritization of contaminated sites for bioremediation, and result in the transfer of improved bioremediation processes for cleanup of contaminated sites.

APPROACH



An important challenge will be to find new ways to investigate molecular and cellular processes under field conditions.


Identifying and understanding the biotransformation and biodegradation of organic and inorganic contaminants will require close integration of laboratory and field experiments. Research areas include aspects of microbial physiology, enzymology, biochemistry, chemistry, microbial ecology, and plant-microorganism interactions. This program element will emphasize metal and radionuclide biotransformations and contaminant biotransformations under field conditions. The role of consortia in contaminant biotransformations will be an explicit component of both laboratory and field investigations. Research carried out under this program element will include all members of soil and water communities, including plants, rhizosphere members, eukaryotes, and prokaryotes. Although the goal of this element is the understanding of biodegradation and biotransformation under field conditions, attaining this understanding will require experiments with pure contaminants and pure cultures under defined conditions. Achieving the goals of this program element will require development of improved experimental laboratory microcosms that qualitatively and quantitatively reflect field mechanisms and processes. The scientific focus of this element is the field and not the laboratory microcosm, and an important challenge will be to find new ways to investigate molecular and cellular processes under field conditions.

Investigations and the expected results will integrate closely with those of the other program elements. Examples of important linkages with the other elements include: identifying rate-limiting enzymatic steps and enzymes as potential targets for the Biomolecular Science and Engineering element; developing the understanding of contaminant transformations required to better assess bioremediation endpoints and performance in the Assessment element; and providing fundamental kinetic data required for the System Integration, Prediction and Optimization element and the Acceleration element.

The Biotransformation and Biodegradation element will focus on four subelements:

1. Biotransformations of metals and radionuclides.

2. Pathways, processes, and molecules for degradation of mixtures of organic contaminants.

3. Interactions between metals and radionuclides, and organics.

4. Kinetics of biotransformation and biodegradation processes under field conditions.

Subelement 1.1: Mechanisms for Biotransformation of Metals and Radionuclides

Fundamental research on those mechanisms that affect the speciation, bioavailability, mobilization, immobilization, and transport of metals and radionuclides in the environment.

Objective


To understand biotransformation of metals and radionuclides in sufficient detail to improve the bioremediation of toxic metals and radionuclides in the environment.

Goals


Three-Year

Identify the dominant redox processes catalyzed by microorganisms and other biota that affect metal speciation, toxicity, and mobility in soils, sediments, and waters. Achieving this goal will involve initial characterization of mechanisms for microbially mediated metal redox reactions and estimation of the importance and potential applicability of these redox reactions in the field. Results obtained will help to identify systems for more detailed study in subsequent years.

Identify and initially characterize biomineralogical processes which affect the mobility and toxicity of toxic metals and radionuclides in soils, sediments, and waters. As with the first goal in this subelement, these results will be used to identify important systems and scientific questions for future study.

Identify those biological processes involving complexation and bioaccumulation of toxic metals and radionuclides that either immobilize or mobilize metals and radionuclides in the environment. This area of research encompasses extracellular metal complexation, complexation to cell surfaces, and intracellular processes for bioaccumulation.

Five-Year

Isolate and characterize enzymes or other molecules responsible for oxidation or reduction of metals and radionuclides and their mobilization or immobilization.

Characterize the role of microbial action on the formation and transport of colloids containing toxic metals and radionuclides in the environment. Because of the multiple mechanisms potentially involved in colloid formation and transport, achieving this goal will require collaboration with other subelements within the Biotransformation and Biodegradation element (complexation with extracellular products and with cell surfaces) and with the Biogeochemical Dynamics element.

Ten-year

Be able to apply the knowledge of biologically mediated redox reactions to improve the management of toxic metal and radionuclide contamination in the environment.

Subelement 1.2: Identification of Pathways, Processes, and Molecules for Degradation of Mixtures of Organic Contaminants

Fundamental research in identifying and characterizing the pathways, processes, and molecules responsible for biotransformations of contaminants that are relevant to field conditions.

Objective


To elucidate the novel and diverse biological mechanisms applicable to degradation of mixtures of organic contaminants, and to identify important biodegradation processes and molecules for other program elements.

Goals


Three-Year

Conceive and initiate investigations of combinations of biological, chemical, and physical processes to degrade mixtures of recalcitrant contaminants in situ or ex situ in reactors. Identify the scientific challenges to application of these methods in the field.

Identify promising targets for biomolecular engineering. Achieving this goal will involve identifying rate-limiting steps in the biodegradation of contaminants and developing methods for identifying and purifying the enzymes or other molecules catalyzing those steps. This goal will include characterization of the catalytic mechanism involved in order to aid in the rational modification of molecules for enhanced activity.

Identify pathway end products and intermediates produced during biodegradation that can be used to measure or assess activity. The purpose is to discover and develop strategies for better measurement of bioremediation activities in the field.

Five-Year

Develop and test one or more strategies, based on fundamental understanding of biodegradation mechanisms, for enhanced biodegradation of organic contaminants in field experimental centers.

Ten-Year

Utilize the understanding of biodegradation mechanisms to improve bioremediation practices and performance in the field.

Subelement 1.3: Interactions between Toxic Metals, Radionuclides, and Organic Contaminants

Fundamental research on the biologically mediated interactions between toxic metals, radionuclides, and organics.

Objective


To understand how and why the presence of toxic metals or radionuclides at a site affects the biodegradation of organics and how biodegradation of contaminants and other organics affects the speciation and mobility of metals and radionuclides.

Goals


Three-Year

Identify and characterize mechanisms whereby biota modify the speciation, mobility, and toxicity of toxic metals and radionuclides during biodegradation of organic contaminants or organic matter. These mechanisms include, for example, the biodegradation of organic complexing agents and the modification of soil organic matter.

Identify and characterize radiological and chemical toxicity and mechanisms for mediation of this toxicity to microorganisms catalyzing beneficial biotransformations of contaminants.

Five-Year

Develop sufficient understanding of the interactions between organic biodegradation, metal and radionuclide toxicity, speciation and mobility, and soil and water biota to test new concepts for control of metal toxicity and mobility in field experimental centers.

Ten-Year

Understand the ways that microorganisms, toxic metals, and radionuclides interact with organic contaminants and organic soil matter to guide bioremediation practices at sites where there are mixtures of metals and organics.

Subelement 1.4: Kinetics of Biotransformation and Biodegradation Processes under Field Conditions

Fundamental research in the kinetics of multi-step, multi-component biodegradation and biotransformation pathways.

Objective


To obtain data describing the rates and extents of contaminant biotransformations relevant to field conditions. These data will be used to develop improved numerical models to elucidate the complex interactions important during bioremediation in the field.

Goals


Three-Year

Initiate studies on the kinetics of coupled biological processes (e.g., cell transport, enzyme transformations of contaminants, cell growth, competition and predation, and feedback inhibition) for the biotransformation of mixtures of contaminants. Although the kinetics of biological processes will be emphasized in this subelement, thermodynamics and reversibility of relevant biotransformations will also be considered. The results of these studies will be used in collaboration with the System Integration element to develop mechanistic models for biodegradation and biotransformation processes at field sites. These studies are directed at identifying (in concert with the Systems Integration, Biogeochemical Dynamics, and Acceleration elements) those processes that have the most value for predicting and improving the performance of bioremediation under field conditions. Therefore, studies of nutrient and substrate limiting conditions, field temperatures, consortia, and other conditions particularly relevant to the field will be emphasized.

Five-Year

Extend investigations of the kinetics of coupled biological processes to field experimental centers and to a more comprehensive collection of contaminants and environmental conditions.

Test the validity, applicability, and ability of the models to predict bioremediation based on the interrelationships of biological, chemical, and physical processes at field experimental centers. Uncertainties of model predictions and sensitivity to accuracies of measured parameters will also be addressed.

Ten-Year

Attain a quantitative and mechanistic understanding of the kinetics and thermodynamics of biodegradation and biotransformation processes to improve bioremediation practices and performance in the field.

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