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| Status |
Public on Apr 30, 2013 |
| Title |
Anaerobic oxidation of benzene by Geobacter metallireducens |
| Organism |
Geobacter metallireducens |
| Experiment type |
Expression profiling by array
|
| Summary |
Anaerobic activation of benzene is expected to represent a novel biochemistry of environmental significance but research into the mechanisms has been stymied by a lack of a genetically tractable pure culture which unequivocally does not use molecular oxygen to activate benzene. Geobacter metallireducens grew in a medium in which benzene was the sole electron donor and Fe(III) was the sole electron acceptor with a stoichiometry of benzene loss and Fe(III) reduction consistent with benzene oxidation to carbon dioxide coupled with Fe(III) reduction. Phenol labeled with 18O was produced when the medium was labeled with H218O, as expected for a true anaerobic conversion of benzene to phenol. Gene expression patterns indicated that benzene was metabolized through a phenol intermediate rather than benzoate or toluene. Deletion of ppcB, which encodes a subunit of the phenylphosphate carboxylase, an enzyme required for phenol metabolism, inhibited metabolism of benzene. Deleting genes specific for benzoate or toluene metabolism did not. Comparison of gene expression patterns in cells grown on benzene versus cells grown on phenol revealed genes specifically expressed in benzene-grown cells. Deletion of one of these, Gmet_3376, inhibited anaerobic benzene oxidation, but not the metabolism of phenol, benzoate, or toluene. The availability of a genetically tractable pure culture that can anaerobically convert benzene to phenol with oxygen derived from water should significantly accelerate elucidation of the mechanisms by which benzene can be activated in the absence of molecular oxygen.
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| Overall design |
Total RNA from
three separate cultures of G. metallireducens grown with 250 µM benzene
three separate cultures of G. metallireducens grown with 500 µM phenol
three separate cultures of G. metallireducens grown with 1 mM benzoate
three separate cultures of G. metallireducens grown with 500 µM toluene
three separate cultures of G. metallireducens grown with 10 mM acetate
were used to study
[1] Anaerobic oxidation of benzene by G. metallireducens (Benzene vs. acetate, Benzene vs. benzoate, Benzene vs. phenol, Benzene vs. toluene)
[2] Anaerobic oxidation of benzoate by G. metallireducens (Benzoate vs. acetate)
[3] Anaerobic oxidation of phenol by G. metallireducens (Phenol vs. acetate)
[4] Anaerobic oxidation of toluene by G. metallireducens (Toluene vs. acetate)
Each chip measures the expression level of 3,627 genes from G. metallireducens DSM 7210 with nine 45-60-mer probe pairs (PM/MM) per gene, with three-fold technical redundancy.
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| Contributor(s) |
Zhang T, Tremblay P, Smith J, Lovley D |
| Citation(s) |
24096430 |
| |
| Submission date |
Nov 17, 2011 |
| Last update date |
Jan 09, 2014 |
| Contact name |
Tian Zhang |
| E-mail(s) |
tzhang@microbio.umass.edu
|
| Phone |
413-577-1745
|
| Organization name |
University of Massachusetts
|
| Department |
Microbiology
|
| Lab |
Derek Lovley's Lab
|
| Street address |
639 North Pleasant St.
|
| City |
Amherst |
| State/province |
MA |
| ZIP/Postal code |
01003 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL14898 |
NimbleGen_Geobacter metallireducens DSM 7210-v1.0 [Design 533273] |
|
| Samples (15)
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| Relations |
| BioProject |
PRJNA148109 |
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