Regulation and genetic organization of hydrogenase: Final progress report for the period June 1, 1985--July 31, 1988

Technical Report · Sat Oct 01 00:00:00 EDT 1988

OSTI ID:6703208

Krasna, A I

Hydrogenase is an enzyme which plays an important role in the anaerobic metabolism of many bacteria. The objectives of the research were to elucidate the regulation and genetic organization of hydrogenase in microorganisms. A mutation in the E. coli hydE gene leads to loss of all hydrogenase activities and isoenzymes as well as all formate-related activities. A 0.9 kb DNA fragment has been cloned from an E. coli genomic DNA library which restored all hydrogenase and formate activities to a hydE mutant strain. This gene coded for a polypeptide of subunit mw 36 kDa which is required for hydrogenase synthesis. It is involved in incorporation of nickel into hydrogenase. A mutation in the E coli hupB gene leads to the loss of the uptake of H/sub 2/ by dyes and the ability to grow on fumarate plus H/sub 2/, but expresses normal levels of hydrogenase when assayed by deuterium exchange. This mutation also leads to loss of all formate-related activities. The mutation mapped near minute 17 and a single mutation was responsible for loss of both activities. A 1.4 kb DNA fragment was isolated which restored the hydrogen uptake activities and coded for a polypeptide of subunit mw 30 kDa. Dna fragments have been isolated from Chromatium vinosum and Proteus vulgaris which restored hydrogenase activities to E. coli strains with mutations in the hydA or hydB regulatory genes and which lack all hydrogenase activities. 6 refs., 12 figs.

• Research Organization: Columbia Univ., New York (USA). Coll. of Physicians and Surgeons
• DOE Contract Number: FG02-85ER13374
• OSTI ID: 6703208
• Report Number(s): DOE/ER/13374-5; ON: DE89003031
• Resource Relation: Other Information: Portions of this document are illegible in microfiche products
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
ESCHERICHIA COLI
METABOLISM
HYDROGENASES
EVALUATION
ANAEROBIC CONDITIONS
DNA-CLONING
FORMATES
GENE REGULATION
GENETIC CONTROL
GENETIC MAPPING
MICROORGANISMS
MUTATIONS
PROGRESS REPORT
BACTERIA
CARBOXYLIC ACID SALTS
CLONING
CONTROL
DNA HYBRIDIZATION
DOCUMENT TYPES
ENZYMES
HYBRIDIZATION
MAPPING
OXIDOREDUCTASES
PEST CONTROL
550200* - Biochemistry
550400 - Genetics