• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. May 1995; 177(10): 2628–2636.
PMCID: PMC176931

Characterization of an operon encoding an NADP-reducing hydrogenase in Desulfovibrio fructosovorans.

Abstract

A genomic DNA fragment from Desulfovibrio fructosovorans, which strongly hybridized with the hydAB genes from Desulfovibrio vulgaris Hildenborough, was cloned and sequenced. This fragment was found to contain four genes, named hndA, hndB, hndC, and hndD. Analysis of the sequence homologies indicated that HndA shows 29, 21, and 26% identity with the 24-kDa subunit from Bos taurus complex I, the 25-kDa subunit from Paracoccus denitrificans NADH dehydrogenase type I, and the N-terminal domain of HoxF subunit of the NAD-reducing hydrogenase from Alcaligenes eutrophus, respectively. HndB does not show any significant homology with any known protein. HndC shows 37 and 33% identity with the C-terminal domain of HoxF and the 51-kDa subunit from B. taurus complex I, respectively, and has the requisite structural features to be able to bind one flavin mononucleotide, one NAD, and three [4Fe-4S] clusters. HndD has 40, 42, and 48% identity with hydrogenase I from Clostridium pasteurianum and HydC and HydA from D. vulgaris Hildenborough, respectively. The 4.5-kb length of the transcripts expressed in D. fructosovorans and in Escherichia coli (pSS13) indicated that all four genes were present on the same transcription unit. The sizes of the four polypeptides were measured by performing heterologous expression of hndABCD in E. coli, using the T7 promoter/polymerase system. The products of hndA, hndB, hndC, and hndD were 18.8, 13.8, 52, and 63.4 kDa, respectively. One hndC deletion mutant, called SM3, was constructed by performing marker exchange mutagenesis. Immunoblotting studies carried out on cell extracts from D. fructosovorans wild-type and SM3 strains, using antibodies directed against HndC, indicated that the 52-kDa protein was recognized in extracts from the wild-type strain only. In soluble extracts from D. fructosovorans wild type, a 10-fold induction of NADP reduction was observed when H(2) was present, but no H(2)-dependent NAD reduction ever occurred. This H(2)-dependent NADP reductase activity disappeared completely in extracts from SM3. These results indicate that the hnd operon actually encodes an NAdP-reducing hydrogenase in D. fructosovorans.

Full Text

The Full Text of this article is available as a PDF (693K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Bruschi M, Guerlesquin F. Structure, function and evolution of bacterial ferredoxins. FEMS Microbiol Rev. 1988 Apr-Jun;4(2):155–175. [PubMed]
  • Fearnley IM, Walker JE. Conservation of sequences of subunits of mitochondrial complex I and their relationships with other proteins. Biochim Biophys Acta. 1992 Dec 7;1140(2):105–134. [PubMed]
  • Fu W, Drozdzewski PM, Morgan TV, Mortenson LE, Juszczak A, Adams MW, He SH, Peck HD, Jr, DerVartanian DV, LeGall J, et al. Resonance Raman studies of iron-only hydrogenases. Biochemistry. 1993 May 11;32(18):4813–4819. [PubMed]
  • Lupton FS, Conrad R, Zeikus JG. Physiological function of hydrogen metabolism during growth of sulfidogenic bacteria on organic substrates. J Bacteriol. 1984 Sep;159(3):843–849. [PMC free article] [PubMed]
  • Nivière V, Wong SL, Voordouw G. Site-directed mutagenesis of the hydrogenase signal peptide consensus box prevents export of a beta-lactamase fusion protein. J Gen Microbiol. 1992 Oct;138(10):2173–2183. [PubMed]
  • Preis D, van der Pas JC, Nehls U, Röhlen DA, Sackmann U, Jahnke U, Weiss H. The 49 K subunit of NADH: ubiquinone reductase (complex I) from Neurospora crassa mitochondria: primary structure of the gene and the protein. Curr Genet. 1990 Jul;18(1):59–64. [PubMed]
  • Rousset M, Dermoun Z, Chippaux M, Bélaich JP. Marker exchange mutagenesis of the hydN genes in Desulfovibrio fructosovorans. Mol Microbiol. 1991 Jul;5(7):1735–1740. [PubMed]
  • Rousset M, Dermoun Z, Hatchikian CE, Bélaich JP. Cloning and sequencing of the locus encoding the large and small subunit genes of the periplasmic [NiFe]hydrogenase from Desulfovibrio fructosovorans. Gene. 1990 Sep 28;94(1):95–101. [PubMed]
  • Rousset M, Dermoun Z, Wall JD, Belaich JP. Analysis of the periplasmic [NiFe] hydrogenase transcription unit from Desulfovibrio fructosovorans. J Bacteriol. 1993 Jun;175(11):3388–3393. [PMC free article] [PubMed]
  • Schneider K, Schlegel HG. Purification and properties of soluble hydrogenase from Alcaligenes eutrophus H 16. Biochim Biophys Acta. 1976 Nov 8;452(1):66–80. [PubMed]
  • Sorokin YI. Role of carbon dioxide and acetate in biosynthesis by sulphate-reducing bacteria. Nature. 1966 Apr 30;210(5035):551–552. [PubMed]
  • Stokkermans J, van Dongen W, Kaan A, van den Berg W, Veeger C. hyd gamma, a gene from Desulfovibrio vulgaris (Hildenborough) encodes a polypeptide homologous to the periplasmic hydrogenase. FEMS Microbiol Lett. 1989 Apr;49(2-3):217–222. [PubMed]
  • Tran-Betcke A, Warnecke U, Böcker C, Zaborosch C, Friedrich B. Cloning and nucleotide sequences of the genes for the subunits of NAD-reducing hydrogenase of Alcaligenes eutrophus H16. J Bacteriol. 1990 Jun;172(6):2920–2929. [PMC free article] [PubMed]
  • Traore AS, Hatchikian CE, Belaich JP, Le Gall J. Microcalorimetric studies of the growth of sulfate-reducing bacteria: energetics of Desulfovibrio vulgaris growth. J Bacteriol. 1981 Jan;145(1):191–199. [PMC free article] [PubMed]
  • Voordouw G, Brenner S. Nucleotide sequence of the gene encoding the hydrogenase from Desulfovibrio vulgaris (Hildenborough). Eur J Biochem. 1985 May 2;148(3):515–520. [PubMed]
  • Voordouw G, Niviere V, Ferris FG, Fedorak PM, Westlake DW. Distribution of Hydrogenase Genes in Desulfovibrio spp. and Their Use in Identification of Species from the Oil Field Environment. Appl Environ Microbiol. 1990 Dec;56(12):3748–3754. [PMC free article] [PubMed]
  • Voordouw G, Strang JD, Wilson FR. Organization of the genes encoding [Fe] hydrogenase in Desulfovibrio vulgaris subsp. oxamicus Monticello. J Bacteriol. 1989 Jul;171(7):3881–3889. [PMC free article] [PubMed]
  • Walker JE. The NADH:ubiquinone oxidoreductase (complex I) of respiratory chains. Q Rev Biophys. 1992 Aug;25(3):253–324. [PubMed]
  • Wierenga RK, Terpstra P, Hol WG. Prediction of the occurrence of the ADP-binding beta alpha beta-fold in proteins, using an amino acid sequence fingerprint. J Mol Biol. 1986 Jan 5;187(1):101–107. [PubMed]
  • Xu XM, Matsuno-Yagi A, Yagi T. Characterization of the 25-kilodalton subunit of the energy-transducing NADH-ubiquinone oxidoreductase of Paracoccus denitrificans: sequence similarity to the 24-kilodalton subunit of the flavoprotein fraction of mammalian complex I. Biochemistry. 1991 Sep 3;30(35):8678–8684. [PubMed]
  • Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. [PubMed]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Conserved Domains
    Conserved Domains
    Link to related CDD entry
  • MedGen
    MedGen
    Related information in MedGen
  • Nucleotide
    Nucleotide
    Published Nucleotide sequences
  • Pathways + GO
    Pathways + GO
    Pathways, annotations and biological systems (BioSystems) that cite the current article.
  • Protein
    Protein
    Published protein sequences
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links
  • Taxonomy
    Taxonomy
    Related taxonomy entry
  • Taxonomy Tree
    Taxonomy Tree

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...