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RAPID DETERMINATION OF MICROBIAL PATHWAYS FOR POLLUTANT DEGRADATION
Impact/Purpose:
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.
Description:
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.