The long-term objective of this project is to acquire new molecular genetic understanding of microbial metabolism of use in collaborative research on the development of improved biotreatment technologies. Targeted compounds are environmentally recalcitrant polyaromatic hydrocarbons and N-heterocyclics (PAH/ NHA), and sulfonated azo dyes. The emphasis is upon aerobic metabolism via cytochrome P450 monooxygenation for the first category, and via oxygen insensitive azo bond cleavage of the azo dyes. Fungal and/or bacterial microorganisms known to carry out these detoxication steps are available, including several that were isolated in other projects of this program. P450 genes will be isolated from microbes processing important monooxygenases and characterized for their enzyme products, by sequence modification and expression in Escherichia coli. Relatively few microbial P450s have been genetically characterized but the available evidence indicates the existence of a large number of highly divergent isomers, present as only a few isomers per microbe. For isolation of P450 genes we propose a possibly general method that targets small segments in the most conserved region of this highly sequence divergent gene superfamily.
Specific Aims 1 and 2: I test two related PCR strategies using the filamentous fungus Cunninghamella elegans and soil mycobacterium strains respectively, for the isolation of P450 genes of PAH/N-heterocyclics metabolism;
Specific Aim 3 : Isolate P450 genes based upon P450 enzymes purified from Cunninghamella sp, as an alternative to Aim 1:
Specific Aim 4 : Isolate and characterize oxygen insensitive azo reductase genes from aerobic Gram negative rod C7. Previously purified enzyme to develop a gene probe;
Specific Aim 5 : Prepare and test pathway using specific gene probes for collaborative studies of microbial communities in biodegradation systems.

Project Start
1999-04-01
Project End
2001-03-31
Budget Start
1998-10-01
Budget End
1999-09-30
Support Year
11
Fiscal Year
2000
Total Cost
$171,067
Indirect Cost
Name
University of Cincinnati
Department
Type
DUNS #
City
Cincinnati
State
OH
Country
United States
Zip Code
45221
McNear Jr, David H; Afton, Scott E; Caruso, Joseph A (2012) Exploring the structural basis for selenium/mercury antagonism in Allium fistulosum. Metallomics 4:267-76
Welsh, Gwendolyn L; Mueller, Kevin E; Soman, Rajiv S et al. (2009) Accessibility of polybrominated diphenyl ether congeners in aging soil. J Environ Monit 11:1658-63
Seo, Youngwoo; Lee, Woo-Hyung; Sorial, George et al. (2009) The application of a mulch biofilm barrier for surfactant enhanced polycyclic aromatic hydrocarbon bioremediation. Environ Pollut 157:95-101
Afton, Scott E; Catron, Brittany; Caruso, Joseph A (2009) Elucidating the selenium and arsenic metabolic pathways following exposure to the non-hyperaccumulating Chlorophytum comosum, spider plant. J Exp Bot 60:1289-97
Chen, Liang; Ovesen, Jerald L; Puga, Alvaro et al. (2009) Distinct contributions of JNK and p38 to chromium cytotoxicity and inhibition of murine embryonic stem cell differentiation. Environ Health Perspect 117:1124-30
Ellis, Jenny; Grimm, Rudolf; Clark, Joseph F et al. (2008) Studying protein phosphorylation in low MW CSF fractions with capLC-ICPMS and nanoLC-CHIP-ITMS for identification of phosphoproteins. J Proteome Res 7:4736-42
Afton, Scott; Kubachka, Kevin; Catron, Brittany et al. (2008) Simultaneous characterization of selenium and arsenic analytes via ion-pairing reversed phase chromatography with inductively coupled plasma and electrospray ionization ion trap mass spectrometry for detection applications to river water, plant extract an J Chromatogr A 1208:156-63
Kubachka, Kevin M; Richardson, Douglas D; Heitkemper, Douglas T et al. (2008) Detection of chemical warfare agent degradation products in foods using liquid chromatography coupled to inductively coupled plasma mass spectrometry and electrospray ionization mass spectrometry. J Chromatogr A 1202:124-31
Seo, Youngwoo; Bishop, Paul L (2008) The monitoring of biofilm formation in a mulch biowall barrier and its effect on performance. Chemosphere 70:480-8
Shah, Monika; Meija, Juris; Caruso, Joseph A (2007) Relative mass defect filtering of high-resolution mass spectra for exploring minor selenium volatiles in selenium-enriched green onions. Anal Chem 79:846-53

Showing the most recent 10 out of 140 publications