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Genetic Engineering of a Radiation-Resistant Bacterium for Biodegradation of Mixed Waste

EPA Grant Number: U915322
Title: Genetic Engineering of a Radiation-Resistant Bacterium for Biodegradation of Mixed Waste
Investigators: Rothfuss, Heather M.
Institution: University of Washington
EPA Project Officer: Thompson, Delores
Project Period: December 1, 1998 through December 1, 2001
Project Amount: $85,112
RFA: STAR Graduate Fellowships (1998)
Research Category: Academic Fellowships , Fellowship - Chemical Engineering , Engineering and Environmental Chemistry

Description:

Objective:

The objectives of this research project are to: (1) clone biodegradative pathways into the highly radiation-resistant bacterium Deinococcus radiodurans that will allow for degradation of organopollutants in the presence of radionuclides; and (2) explore the degradative capability of recombinant strains in mixed wastes.

Approach:

Organisms that have the ability to degrade a broad spectrum of organics or convert them to nontoxic intermediates are ubiquitous in nature. Several organisms that can remediate heavy metals or protect themselves from being harmed by heavy metals also have been isolated. Many of the enzymes implicated in these abilities have been isolated and genetically characterized. However, in the presence of moderate- to high-level ionizing radiation, most organisms die or mutate because of the inability to quickly and correctly repair strand breaks caused by the radiation. The nonpathogenic bacterium D. radiodurans is an exception. The Deinococci have a very quick and accurate DNA repair system, can withstand ionizing radiation doses in the 5 Mrad range, and show no loss of viability at 500 krad. However, Deinococci do not appear to have a natural ability to degrade toxic organics. In this research project, our approach is to select enzymes that have the potential to allow D. radiodurans to attack organics of concern, to clone the genes encoding these enzymes into the chromosome of the bacterium, and to evaluate the level of expression, the stability, and the effect of the genes on D. radiodurans. This information will be used to formulate possible treatment strategies using the recombinant bacterium.

Supplemental Keywords:

fellowship, Deinococcus radiodurans R1, bioremediation, mixed waste, oxygenase, toxic organic, genetic engineering, metabolism, enzymes, carcinogen, mutagen, solvents, dense nonaqueous phase liquid, DNAPL, nonaqueous phase liquid, NAPL, bacteria, oxidation, waste reduction, recombinant bacterium, DNA, Deinococci, organopollutants, radionuclides. , POLLUTANTS/TOXICS, TREATMENT/CONTROL, Scientific Discipline, Waste, Chemical Engineering, Chemicals, Biochemistry, Environmental Engineering, Environmental Chemistry, Treatment Technologies, Bioremediation, Genetics, heavy metals, biodegradation, radiation resistant bacterium, genetic engineering, cloning, genetically engineered microorganisms

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The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.

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