Natural nitro compounds with diverse structures and biological functions are produced by bacteria, fungi, plants and animals. Little is known about the behavior of such compounds in natural ecosystems. The lack of accumulation in the biosphere implies that they are biodegraded. Microbial strategies for biodegradation of synthetic nitro compounds are well established; however only two pathways are known for degradation of natural nitro compounds. The research described here examines the genes that encode the key enzymes required for biodegradation of 3-nitrotyrosine (3NTyr) and 1-nitro-2-phenylethane (NPE).
3NTyr is a biological marker for disease and inflammation in plants and animals. A 3NTyr degrading microbe, Variovorax sp. JS669 was isolated from soil. We identified the JS669 denA, which encodes an enzyme that catalyzes denitration of 4-hydroxy-3-nitro-phenylacetate, the key step in metabolism of 3NTyr. The isolation of 3NTyr degraders and development of molecular probes specific to denA revealed that 3NTyr degradation is a widespread phenomena in natural habitats and the compound is metabolized by phylogenetically diverse bacteria. Phylogenetic analysis of the 4-hydroxy-3-nitro-phenylacetate denitrase from JS669 revealed it to be the first functionally annotated protein in a clade of unidentified Class A flavin monooxygenases.
NPE has been identified from several plants, yet the biodegradation of the compound remained a mystery. Here we report the degradation of NPE and its analog 2-nitropropylbenzene. Discovery of the metabolic pathway revealed a novel microbial strategy to use a meta-ring fission degradation pathway to cleave an undesirable side chain from an aromatic compound and use the remainder of the compound as a carbon and energy source. Two genes that encode enzymes in the biodegradation pathway were identified and both are deeply branched within their respective phylogenetic trees, indicating that both represent highly specialized microbial enzymes. Furthermore, microbial degradation of NPE resulted in the production of 3-nitropropionic acid, a natural toxin that inhibits succinate dehydrogenase and is responsible for livestock illness and death. This is the first report of bacterial production of 3-nitropropionic acid, and might represent a significant source of 3-nitropropionic acid in natural habitats.
The findings from these studies contribute to the overall understanding of microbial metabolism. Specifically, this research reveals genes that encode novel enzymes and strategies for the biodegradation of two natural nitro compounds. Furthermore, discovery of mechanisms for the biodegradation of such compounds reveals novel microbial metabolic diversity and provides insight into the evolution of degradation pathways for synthetic compounds.
| Identifer | oai:union.ndltd.org:GATECH/oai:smartech.gatech.edu:1853/34002 |
| Date | 06 April 2010 |
| Creators | Parks, Samantha Terris |
| Publisher | Georgia Institute of Technology |
| Source Sets | Georgia Tech Electronic Thesis and Dissertation Archive |
| Detected Language | English |
| Type | Dissertation |
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