Cloning and sequencing of a [NiFe] hydrogenase operon from Desulfovibrio vulgaris Miyazaki F
- 1 October 1990
- journal article
- research article
- Published by Microbiology Society in Journal of General Microbiology
- Vol. 136 (10) , 2021-2028
- https://doi.org/10.1099/00221287-136-10-2021
Abstract
A hydrogenase operon was cloned from chromosomal DNA isolated from Desulfovibrio vulgaris Miyazaki F with the use of probes derived from the genes encoding [NiFe] hydrogenase from Desulfovibrio vulgaris Hildenborough. The nucleic acid sequence of the cloned DNA indicates this hydrogenase to be a two-subunit enzyme: the gene for the small subunit (267 residues; molecular mass = 28763 Da) precedes that for the large subunit (566 residues; molecular mass = 62495 Da), as in other [NiFe] and [NiFeSe] hydrogenase operons. The amino acid sequences of the small and large subunits of the Miyazaki hydrogenase share 80% homology with those of the [NiFe] hydrogenase from Desulfovibrio gigas. Fourteen cysteine residues, ten in the small and four in the large subunit, which are thought to co-ordinate the iron-sulphur clusters and the active-site nickel in [NifFe] hydrogenases, are found to be conserved in the Miyazaki hydrogenase. The subunit molecular masses and amino acid composition derived from the gene sequence are very similar to the data reported for the periplasmic, membrane-bound hydrogenase isolated by Yagi and coworkers, suggesting that this hydrogenase belongs to the general class of [NiFe] hydrogenases, despite its low nickel content and apparently anomalous spectral properties.This publication has 26 references indexed in Scilit:
- Cloning of the gene encoding the hydrogenase from Desulfovibrio vulgaris (Hildenborough) and determination of the NH2-terminal sequenceEuropean Journal of Biochemistry, 1985
- Nucleotide sequence of the gene encoding the hydrogenase from Desulfovibrio vulgaris (Hildenborough)European Journal of Biochemistry, 1985
- Analysis of the Active Center of Hydrogenase from Desulfovibrio vulgaris Miyazaki by Magnetic Measurements1The Journal of Biochemistry, 1985
- A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragmentsGene, 1982
- The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primersGene, 1982
- Unambiguous identification of the nickel EPR signal in 61Ni-enriched Desulfovibrio gigas hydrogenaseBiochemical and Biophysical Research Communications, 1982
- Characterization of the periplasmic hydrogenase from Desulfovibrio gigasBiochemical and Biophysical Research Communications, 1978
- DNA sequencing with chain-terminating inhibitorsProceedings of the National Academy of Sciences, 1977
- Properties of Purified Hydrogenase from the Particulate Fraction of Desulfovibrio vulgaris, Miyazaki1The Journal of Biochemistry, 1976
- A procedure for the isolation of deoxyribonucleic acid from micro-organismsJournal of Molecular Biology, 1961