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Title: Thiophene metabolism by E. coli

Abstract

The objective of this project is to investigate the mechanism of degradation of sulfur containing heterocyclic molecules by mutants of Escherichia coli K-12. We previously isolated multiple mutants of E. coli which were selected for improved oxidation of furan and thiophene derivatives. We have focused on the thdA mutation in our subsequent research as it appears to be of central importance in thiophene oxidation. We hope that analysis of the thd gene of E. coli will lead to improvement of our thiophene metabolizing bacterial strains.

Authors:
Publication Date:
Research Org.:
Southern Illinois Univ., Carbondale, IL (USA). Dept. of Microbiology
Sponsoring Org.:
DOE/FE
OSTI Identifier:
6296900
Report Number(s):
DOE/PC/79912-13
ON: DE91005924
DOE Contract Number:
FG22-87PC79912
Resource Type:
Technical Report
Country of Publication:
United States
Language:
English
Subject:
01 COAL, LIGNITE, AND PEAT; 59 BASIC BIOLOGICAL SCIENCES; COAL; DESULFURIZATION; THIOPHENE; BIODEGRADATION; CULTURE MEDIA; ESCHERICHIA COLI; GENES; MUTANTS; PROGRESS REPORT; SULFONES; BACTERIA; CARBONACEOUS MATERIALS; CHEMICAL REACTIONS; DECOMPOSITION; DOCUMENT TYPES; ENERGY SOURCES; FOSSIL FUELS; FUELS; HETEROCYCLIC COMPOUNDS; MATERIALS; MICROORGANISMS; ORGANIC COMPOUNDS; ORGANIC SULFUR COMPOUNDS; 010402* - Coal, Lignite, & Peat- Purification & Upgrading; 550700 - Microbiology

Citation Formats

Clark, D.P.. Thiophene metabolism by E. coli. United States: N. p., 1990. Web. doi:10.2172/6296900.
Clark, D.P.. Thiophene metabolism by E. coli. United States. doi:10.2172/6296900.
Clark, D.P.. 1990. "Thiophene metabolism by E. coli". United States. doi:10.2172/6296900. https://www.osti.gov/servlets/purl/6296900.
@article{osti_6296900,
title = {Thiophene metabolism by E. coli},
author = {Clark, D.P.},
abstractNote = {The objective of this project is to investigate the mechanism of degradation of sulfur containing heterocyclic molecules by mutants of Escherichia coli K-12. We previously isolated multiple mutants of E. coli which were selected for improved oxidation of furan and thiophene derivatives. We have focused on the thdA mutation in our subsequent research as it appears to be of central importance in thiophene oxidation. We hope that analysis of the thd gene of E. coli will lead to improvement of our thiophene metabolizing bacterial strains.},
doi = {10.2172/6296900},
journal = {},
number = ,
volume = ,
place = {United States},
year = 1990,
month = 1
}

Technical Report:

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  • The objective of this project is to investigate the mechanism of thiophene and furan degradation by E. coli and to characterize the genes involved and the enzyme activities of their products. 4 tabs.
  • The objective of this project is to investigate the mechanism of degradation of sulfur containing heterocyclic molecules such as those found in coal, by mutants of Escherichia coli K-12. We previously isolated multiple mutants of E. coli which were selected for improved oxidation of furan and thiophene derivatives. We have focused on the thdA mutation in our subsequent research as it appears to be of central importance in thiophene oxidation. We hope that analysis of the thd genes of E. coli will lead to improvement of our thiophene metabolizing bacterial strains. 1 tab.
  • The objective for this project was to continue the genetic analysis of the thiophene oxidation system, in particular: 1. characterization of the biochemical pathway for thiophene oxidation, 2. identification and mapping of any further genes involved in thiophene degradation, 3. analysis of how the thd genes are regulated, and 4. cloning and sequencing of at least some of the thd genes.
  • The objective of this project is to investigate the mechanism of degradation of sulfur-containing heterocyclic molecules by mutant strains of Escherichia coli K-12. We have previously isolated multiple mutants of E. coli which had gained the capacity to oxidize thiophene compounds and their furan analogs. We have focused on the thdA mutation in our subsequent research, as this appears to be in a regulatory gene central to the thiophene/furan oxidation system. The thdF gene appears to be more directly involved in the oxidation reactions, whereas thdC and thdD are apparently required for increased protection against the toxic effects of thiophenemore » and furan compounds. 4 tabs.« less
  • The objective of this project is to investigate the mechanism of thiophene and furan degradation by mutants of Escherichia coli K-12 which have gained the ability to use such heterocyclic compounds. We hope to characterize the genes involved in these novel breakdown pathways and, if possible to assay the activities of the corresponding activities. Previous work resulted in the isolation of a series of mutants of E. coli with increasing ability to oxidize furan and thiophene derivatives. Mutations in three novel genes were involved, thdA, thdC and thdD together with mutations in fadR (''thdB'') and atoC. Of these thdA gavemore » the biggest observable effect and we have therefore focused on this gene in our more recent work. Although the nature of the thdA system is still unknown, it is clearly rather non-specific (as is P450) and can be mutated to various mutant forms with different specificities. Lastly, we isolated novel furan degrading mutant starting from a different genetic background. These mutants contrast with those made from DC625 in that they could use furans and benzoic acid as sole carbon and energy source implying complete degradation. The new mutants showed moderate activity against some thiophene derivatives but not others. These mutants are very interesting and will be further analysed both genetically and for their biochemical properties.« less