Molecular biology of coal bio-desulfurization

Technical Report · Thu Jul 18 00:00:00 EDT 1991

OSTI ID:5446524

Young, K D; Gallagher, J R

The aim of this project is to use the techniques of molecular genetics to identify, clone, sequence, and enhance the expression of proteins which remove sulfur covalently bound to coal. The cloned dox genes from strain C18 were more fully characterized. Another gene, doxG, is almost identical with the nahC gene, also involved in the naphthalene pathway. These results lead us to believe that dibenzothiophene (DBT) is degraded by the oxidative route along the naphthalene degradative pathway. At least one other gene, doxI, is implicated in the formation of the unidentified fluorescent product from DBT. The Rhodococcus rhodochrous isolate IGTS8 was mutated to produce a variant (strain UV1) that was negative for DBT utilization. This mutant was exposed to further rounds of UV mutagenesis and was used as the host for attempts to introduce various plasmid DNAs. Two plasmids could be introduced into UV1: pRF29 and pLAFR5. A cosmid library of IGTS8 DNA was constructed in pLAFR5 and was electroporated into UV1. Of over 2000 colonies tested three bulk batches, one flask of about 600 clones produced a fluorescent product from DBT. These are being screened to determine if one of these represents a clone that complements the DBT negative phenotype of the UV1 mutant. We obtained chloramphenicol resistant Thiobacillus ferrooxidans after electroporation but these transformants contained no plasmid or transposon sequences, so that insertion of DNA into T. ferrooxidans had not occurred.

• Research Organization: North Dakota Univ., Grand Forks, ND (United States). Dept. of Microbiology and Immunology
• Sponsoring Organization: USDOE; USDOE, Washington, DC (United States)
• DOE Contract Number: AC22-89PC89901
• OSTI ID: 5446524
• Report Number(s): DOE/PC/89901-T5; ON: DE91018159
• Country of Publication: United States
• Language: English

Related Subjects

01 COAL, LIGNITE, AND PEAT
59 BASIC BIOLOGICAL SCIENCES
COAL
DESULFURIZATION
THIONAPHTHENES
BIODEGRADATION
CLONING
GENES
MICROORGANISMS
MUTATIONS
PLASMIDS
PROGRESS REPORT
PROTEINS
THIOBACILLUS FERROXIDANS
BACILLUS
BACTERIA
CARBONACEOUS MATERIALS
CELL CONSTITUENTS
CHEMICAL REACTIONS
DECOMPOSITION
DOCUMENT TYPES
ENERGY SOURCES
FOSSIL FUELS
FUELS
HETEROCYCLIC COMPOUNDS
MATERIALS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
SULFUR-OXIDIZING BACTERIA
010402* - Coal
Lignite
& Peat- Purification & Upgrading
550700 - Microbiology
550400 - Genetics