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Purification and characterization of a membrane-bound hydrogenase from the hyperthermophilic archaeon Pyrococcus furiosus


, : Purification and characterization of a membrane-bound hydrogenase from the hyperthermophilic archaeon Pyrococcus furiosus. Journal of Bacteriology 182(12): 3423-3428

Highly washed membrane preparations from cells of the hyperthermophilic archaeon Pyrococcus furiosus contain high hydrogenase activity (9.4 micromol of H(2) evolved/mg at 80 degrees C) using reduced methyl viologen as the electron donor. The enzyme was solubilized with n-dodecyl-beta-D-maltoside and purified by multistep chromatography in the presence of Triton X-100. The purified preparation contained two major proteins (alpha and beta) in an approximate 1:1 ratio with a minimum molecular mass near 65 kDa and contained approximately 1 Ni and 4 Fe atoms/mol. The reduced enzyme gave rise to an electron paramagnetic resonance signal typical of the so-called Ni-C center of mesophilic NiFe-hydrogenases. Neither highly washed membranes nor the purified enzyme used NAD(P)(H) or P. furiosus ferredoxin as an electron carrier, nor did either catalyze the reduction of elemental sulfur with H(2) as the electron donor. Using N-terminal amino acid sequence information, the genes proposed to encode the alpha and beta subunits were located in the genome database within a putative 14-gene operon (termed mbh). The deduced sequences of the two subunits (Mbh 11 and 12) were distinctly different from those of the four subunits that comprise each of the two cytoplasmic NiFe-hydrogenases of P. furiosus and show that the alpha subunit contains the NiFe-catalytic site. Six of the open reading frames (ORFs) in the operon, including those encoding the alpha and beta subunits, show high sequence similarity (>30% identity) with proteins associated with the membrane-bound NiFe-hydrogenase complexes from Methanosarcina barkeri, Escherichia coli, and Rhodospirillum rubrum. The remaining eight ORFs encode small (<19-kDa) hypothetical proteins. These data suggest that P. furiosus, which was thought to be solely a fermentative organism, may contain a previously unrecognized respiratory system in which H(2) metabolism is coupled to energy conservation.

US$19.90

PMID: 10852873

DOI: 10.1128/JB.182.12.3423-3428.2000


Other references

Dugin, S.F.; Medvedev, O.S.; Glukhovtsev, E.V.; Khodzhaev, A.I., 1989: Changes in systemic and regional hemodynamics in the acute period of myocardial infarction in conscious rabbits. Systemic and regional hemodynamics (radioactive microspheres of 15 mcm) were studied in 7 conscious immobilized rabbits 30 min, 24 and 72 hours after left coronary artery occlusion (group 1) and 30 min and 24 hours after the occlusion in 5 rabbits...

Tang, P.Hwee.; Ong, C.Li.; Stringer, D.; Tan, J.V.K.; Yeo, G.S.H., 2010: Magnetic resonance imaging of the fetal central nervous system in Singapore. Fetal imaging has improved with the development of faster magnetic resonance imaging (MRI) sequences, obviating the requirement for sedation. It is useful in characterising abnormality of the central nervous system in fetuses with abnormal or equi...

Pinto, M.G. de S., 1996: Single entry volume tables for Betula sp., Castanea crenata, Quercus robur and Quercus rubra. Based on data collected from temporary sampling plots located in the forestry areas of Serra da Cabreira, Senhora da Abadia and Entre Vez e Coura, Portugal, local volume tables (over and under bark) were prepared for these 4 species. Volume estima...

Yoshikura, H., 1973: Ultraviolet inactivation of murine leukemia virus and its assay in permissive and non-permissive cells. International Journal of Cancer 11(3): 739-746

Gambi G., 1983: Conclusions and proposals. In this conclusive note the results of the different studies on black pine stands in the central-northern Apennines [Italy] are summarized. The problem of thinning, both in young stands at initial spacing and in grown-up unthinned ones, is conside...

Silfverskiöld, B.P., 1968: Northern Physicians' Committee on Medical Injuries in Boxing. No prohibition now but new safety injunctions. Lakartidningen 65(6): 583-587

Anonymous, 2004: Even when walking yet does not cause any pain, peripheral arterial occlusive disease costs life years. Mmw Fortschritte der Medizin 145(15): 63-63

Zamponi, G.W., 2009: Launching Channels: a new milestone in the ion channel field. Channels 1(1): 1-1

Lee, E.D.; Lee, S.D.; Kim, W.B.; Yang, J.; Kim, S.H.; Lee, M.G., 1996: Determination of a new carbapenem antibiotic, DA-1131, in rat plasma, urine, and bile by column-switching high-performance liquid chromatography. A column-switching high-performance liquid chromatographic method has been developed for the determination of a new carbapenem antibiotic, DA-1131, in rat plasma, urine, and bile. Each biological sample was diluted with an equal volume of a stabil...

Veen, P.V.n-Der; Voragen, A.; Visser, J., 1990: Production, structure and function of some arabinases produced by Aspergillus niger. Agricultural biotechnology in focus in the Netherlands JJ Dekkers HC van der Plas and DH Vuijk eds: 192