Genetic engineering of sulfur-degrading Sulfolobus

Technical Report · Mon Dec 31 23:00:00 EST 1990

OSTI ID:5568694

Ho, N W.Y. ^[1]

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1. Purdue Univ., Lafayette, IN (USA). Lab. of Renewable Resources Engineering

Recent studies have shown that some microorganisms can play a significant role in removing the sulfur compound from coal. Sulfolobus acidocaldarius and related species are such microorganisms. The objective of this project is to develop a genetic transformation system for Sulfolobus species so that they could become the ideal host to overproduce homologous and heterologous enzymes that are most effective for the removal of sulfur from coal, particularly organic sulfur. Last quarter, we have identified three chemicals that can inhibit the growth of S. Acidocaldarius. These chemicals can be part of the selection system for the development of a transformation system for S. acidocaldarius. Due to the fact that Sulfolobus shibatae B12 becomes increasingly more attractive as a host for housing genes encoding desulfurization enzymes, in this period we also studied the affect of these three chemicals to growth of S. shibatae B12. We found that S. shibatae B12 is also sensitive to these chemicals. This quarter we succeeded in the isolation and purification of the double-stranded DNA virus from S. shibatae B12. Furthermore, the individual EcoRI and BamH1 fragments of the virus have also been cloned into pUC19 plasmid. These plasmids will be used for the construction of the final E. coli-Sulfolobus shuttle vector. 5 Flurouracil (5FU) is one of the chemicals that inhibit growth of Sulfolobus. Resistance strain of S. acidocaldarius to 5FU has also been isolated. DNA from the 5FU resistance strain has also been isolated. 2 figs.

• Research Organization: Illinois Dept. of Energy and Natural Resources, Springfield, IL (USA)
• Sponsoring Organization: DOE; ILDENR; USDOE, Washington, DC (USA); Illinois Dept. of Energy and Natural Resources, Springfield, IL (USA)
• DOE Contract Number: FG22-90PC90176
• OSTI ID: 5568694
• Report Number(s): DOE/PC/90176-T24; ON: DE91014050
• Country of Publication: United States
• Language: English

Related Subjects

01 COAL, LIGNITE, AND PEAT
010402* -- Coal
Lignite
& Peat-- Purification & Upgrading
550400 -- Genetics
550700 -- Microbiology
59 BASIC BIOLOGICAL SCIENCES
BIOCONVERSION
CARBONACEOUS MATERIALS
CELL CONSTITUENTS
CHEMICAL REACTIONS
COAL
DESULFURIZATION
DNA
DOCUMENT TYPES
ENERGY SOURCES
ENZYMES
FOSSIL FUELS
FUELS
GENETIC ENGINEERING
MATERIALS
MICROORGANISMS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PLASMIDS
PROGRESS REPORT
RECOMBINANT DNA