Rapid Determination of Microbial Pathways for Pollutant Degradation

EPA Grant Number: R831276C011
Subproject: this is subproject number 011 , established and managed by the Center Director under grant CR831276
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).

Center: Gulf Coast HSRC (Lamar)
Center Director: Ho, Tho C.
Title: Rapid Determination of Microbial Pathways for Pollutant Degradation
Investigators: Kinney, Kerry A. , Whitman, Christian
Current Investigators: Kinney, Kerry A. , Kirisits, Mary Jo , Whitman, Christian
Institution: The University of Texas at Austin
EPA Project Officer: Lasat, Mitch
Project Period: December 1, 2003 through November 30, 2004
Project Amount: Refer to main center abstract for funding details.
RFA: Gulf Coast Hazardous Substance Research Center (Lamar University) (1996) RFA Text |  Recipients Lists
Research Category: Hazardous Waste/Remediation , Targeted Research

Objective:

Significant quantities of organic pollutants are released into the air, soils and waterways of the Gulf Coast Region posing a potential health risk to nearby communities. These pollutants are a drain on our economy as investigation and clean up costs can be prohibitive. Both in situ and ex situ biological treatment processes are an attractive control option for organic pollutants because they are cost effective, consume little energy, and can convert organic pollutants into benign products. Despite the promise of these methods, controlling and optimizing these systems to the field can be difficult due to their inherent complexity. In addition, current microbiological and analytical methods for determining biodegradation pathways are cumbersome and time consuming for the poorly characterized microorganisms usually found in treatment systems. A diagnostic tool that would allow engineers to more rapidly determine how the design and operation of their biotreatment system affects key biodegradation pathways would greatly enhance our ability to quickly optimize pollutant biodegradation rates.

The objective of the proposed research is therefore to develop a molecular tool, complementary DNA (cDNA) subtraction, in a novel way that allows rapid and efficient determination of the metabolic pathways used by microorganisms to degrade organic pollutants. The cDNA subtraction technique is well suited for environmental applications since it can identify specifically those genes that are expressed for pollutant degradation and does not require a priori knowledge of a microorganism's metabolism.

Approach:

Specific tasks of the proposed project include the following:

  • Develop the cDNA subtraction tool for application to environmentally relevant bacteria and verify that it can rapidly identify the genes expressed in the biodegradation of a model organic compound.
  • Develop an internal "standard addition" control method to quantify the efficiency of the cDNA subtraction method so that it can be applied to uncharacterized bacteria.
  • Assess the capability of the cDNA subtraction tool to determine the pollutant degradation pathways in a relatively uncharacterized bacterial species.
  • Use the gene sequence information obtained in the steps above to quantify gene expression patterns in microbial cultures degrading BTEX mixtures. It is our intent to verify that the cDNA subtraction tool and focused gene expression studies can be used to efficiently determine regulation patterns in key biodegradation pathways.
  • Examine the utility of the cDNA subtraction tool and focused gene expression studies to assess the biodegradation potential of the mixed microbial cultures typically found in biological treatment systems.

    Expected Results:

    The proposed project period is three years and the total GCHSRC funding requested is as follows: $49,811 (Year 1), $49,555 (Year 2) and $49,917 (Year 3). The final product of this research effort will be a cDNA subtraction tool which has been developed specifically for delineating biodegradation pathways utilized in environmental treatment applications. Rather than having to sequence the whole genome of a particular microbial species (a multiyear effort), the cDNA subtraction method allows one to delineate those genes expressed only during the biodegradation of a particular pollutant. Since this effort is much more focused, it allows one to identify the environmentally relevant genes in a matter of days or weeks. This information lays the foundation for efficient development of molecular probes to monitor gene expression in ex situ or in situ biological treatment systems. The ultimate cost savings provided by the cDNA subtraction method coupled with focused gene expression will likely be site and treatment system specific. However, if one considers the hundreds of thousands (to multimillions) of dollars that are often spent to investigate and remediate a single hazardous waste site, successful deployment of this molecular tool has the potential to provide significant cost savings.

    Publications and Presentations:

    Publications have been submitted on this subproject: View all 4 publications for this subprojectView all 64 publications for this center

    Supplemental Keywords:

    RFA, Scientific Discipline, INTERNATIONAL COOPERATION, Waste, TREATMENT/CONTROL, Remediation, Environmental Chemistry, Treatment Technologies, Technology, Hazardous Waste, Hazardous, hazardous waste treatment, microbial degradation, BTEX, biotechnology, cDNA subtraction tool, bioremediation

    Progress and Final Reports:

  • Final

  • Main Center Abstract and Reports:

    CR831276    Gulf Coast HSRC (Lamar)

    Subprojects under this Center: (EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
    R831276C001 DNAPL Source Control by Reductive Dechlorination with Fe(II)
    R831276C002 Arsenic Removal and Stabilization with Synthesized Pyrite
    R831276C003 A Large-Scale Experimental Investigation of the Impact of Ethanol on Groundwater Contamination
    R831276C004 Visible-Light-Responsive Titania Modified with Aerogel/Ferroelectric Optical Materials for VOC Oxidation
    R831276C005 Development of a Microwave-Induced On-Site Regeneration Technology for Advancing the Control of Mercury and VOC Emissions Employing Activated Carbon
    R831276C006 Pollution Prevention through Functionality Tracking and Property Integration
    R831276C007 Compact Nephelometer System for On-Line Monitoring of Particulate Matter Emissions
    R831276C008 Effect of Pitting Corrosion Promoters on the Treatment of Waters Contaminated with a Nitroaromatic Compounds Using Integrated Reductive/Oxidative Processes
    R831276C009 Linear Polymer Chain and Bioengineered Chelators for Metals Remediation
    R831276C010 Treatment of Perchlorate Contaminated Water Using a Combined Biotic/Abiotic Process
    R831276C011 Rapid Determination of Microbial Pathways for Pollutant Degradation
    R831276C012 Simulations of the Emission, Transport, Chemistry and Deposition of Atmospheric Mercury in the Upper Gulf Coast Region
    R831276C013 Reduction of Environmental Impact and Improvement of Intrinsic Security in Unsteady-state
    R831276C014 Integrated Chemical Complex and Cogeneration Analysis System: Greenhouse Gas Management and Pollution Prevention Solutions
    R831276C015 Improved Combustion Catalysts for NOx Emission Reduction
    R831276C016 A Large-Scale Experimental Investigation of the Impact of Ethanol on Groundwater Contamination
    R831276C017 Minimization of Hazardous Ion-Exchange Brine Waste by Biological Treatment of Perchlorate and Nitrate to Allow Brine Recycle
    R831276C018 Integrated Chemical Complex and Cogeneration Analysis System: Greenhouse Gas Management and Pollution Prevention Solutions