133363347.

doc

BIO 220 7. Bioremediation Bioremediation- (Biodegradation Technologies) Bioremediation is the process by which bacteria and some Eukaryotic microorganisms are used to clean up polluted sites or improve current industrial processes. Bioremediation is the utilization of microbes naturally occurring catabolic pathways. Originally bioremediation involved non-modified organisms, but limited studies have now occurred involving genetically engineered microorganisms. Advantages of Bioremediation Moderate capital investment Nontoxic Biodegradable Relatively self-sustaining Variety of approaches available In Situ vs. Ex Situ Bioremediative process can occur on-site (in situ) or at off-site locations (ex situ). In situ approaches are preferred due to lower costs involved. However in-situ bioremediation is not always possible.

Degradation of toxic compoundsHydrocarbons Chlorinated compounds Heavy Metals Improvement of Industrial Processes Removal of aphid secretions on cotton plants Removal of dyes from effluent wastewater Hydrocarbon Transformation Pseudomonas sp. Aliphatic and Aromatic compounds Oxygen added to compounds Monooxygenase Dioxygenase Series of oxidation/reduction reactions coverts HC into acetylCoA
1

133363347.doc

Acetyl-CoA feeds directly into Krebs Cycle! Must occur in aerobic environment O2 or H2O2 directly pumped into anoxic sediments-Bioventing Addition of PO4/NO3 to stimulate growth Exxon Valdez oil spill

Chlorinated Compounds Chlorinated compounds are common industrial solvents, degreasers and dry cleaning agents which often cause cancer. Chlorinated compounds are a significant cause of groundwater contamination in D.O.D. clean-up sites. Reductive Dechlorination is the process by which chlorine groups are removed and replaced with hydrogen. Ralstonia eutropha is a well studied organism capable of Reductive Dechlorination. Organism was first isolated from contaminated sediments in Hudson River. Variety of chlorinated compoundsPCBs (polychlorinated biphenyls) TCA (trichloroethane) TCE (trichloroethylene) PCBs Multiple chlorination sites Ortho and Meta confirmations more difficult to degrade Multiple chlorinations best degraded anaerobically Limited chlorination sites best degraded aerobically Deholabacter restrictus Acetate as carbon source H2 as electron donor TCA as electron acceptor Mercury Cleanup Ionic Mercury (Hg++) was a common ingredient in thermometers, tooth filings and frequently used in industrial processes. Mercury poisoning usually concentrates in organisms higher on the food chain (Biomagnification). Mercury spewed from factories in the NE USA has precipitated in rain at levels higher than regarded safe by the EPA! Microbes resistant to mercury have been characterized. Genes in Mer operon allow the organism to reduce mercury to the less toxic form of
2

133363347.doc

Hg. Organisms able to reduce organic mercury (such as methyl mercury) are said to have Broad Spectrum Resistance to mercury. Organisms that can only reduce inorganic mercury (Hg++) are said to possess Narrow Spectrum Resistance. The Mer Operon can be found on plasmids, transposons and the bacterial chromosome. The operon ahs now been engineered into E. coli, a bacteria not normally resistant to mercury. Mechanism for mercury detoxification Periplasmic binding protein Mer P binds Hg++ on cysteine residues Transfers to cysteine residues on cytoplasmic protein Mer T Hg++ is transferred to NADPH dependent Mercuric Reductase Hg++ is reduced to nontoxic Hg0 Regulation of expression of genes in Mer Operon is controlled by a regulatory protein Mer R. In the presence of Hg++ Mer R allows induction of transcription whereas in the absence of Hg++ the protein acts as a repressor.

Selenium Cleanup Selenium (Se) is a heavy metal which has concentrated to toxic levels in the southern portion of Californias Central Valley. Enterobacter cloacae can reduce toxic selenium trioxide (SeO42-) to metallic Selenium (Se0). Genetically Engineered Bioremediative Agents As organisms possessing bioremediative capabilities have been characterized, scientists have taken steps to transfer responsible genes into other organisms. As naturally occurring organisms may have habitat limitations and slower growth rates, there is considerable interest in these molecular approaches. Organisms can be modified in several ways: Substrate specificity Pathway modification Toxicity assessment (bioluminescence) Gene Transfer occurs using: Conjugative plasmids Plasmid + electroporation Transposon derived gene cassettes
3

133363347.doc

Although genetically modified organisms have shown success in bioremediative trials in confined environments, the EPA has yet to allow deliberate release of genetically modified organisms for the purpose of bioremediation. Biopolymers To reduce the amount of synthetic plastics in landfills, a movement is underway to identify biodegradable alternatives- biopolymers. Biopolymers include starch based plastics and copolymers such as PHV/PHB containing equal amounts of -hydroxyvalerate and hydroxybutyrate. (See Figure 19.49) Ultraviolet light from the sun breaks down the polymers into forms easily susceptible to microbial attack.