Abstract:
:The fermentative hyperthermophile Pyrococcus furiosus contains an NADPH-utilizing, heterotetrameric (alphabetagammadelta), cytoplasmic hydrogenase (hydrogenase I) that catalyzes both H(2) production and the reduction of elemental sulfur to H(2)S. Herein is described the purification of a second enzyme of this type, hydrogenase II, from the same organism. Hydrogenase II has an M(r) of 320,000 +/- 20,000 and contains four different subunits with M(r)s of 52,000 (alpha), 39,000 (beta), 30,000 (gamma), and 24,000 (delta). The heterotetramer contained Ni (0.9 +/- 0.1 atom/mol), Fe (21 +/- 1.6 atoms/mol), and flavin adenine dinucleotide (FAD) (0.83 +/- 0.1 mol/mol). NADPH and NADH were equally efficient as electron donors for H(2) production with K(m) values near 70 microM and k(cat)/K(m) values near 350 min(-1) mM(-1). In contrast to hydrogenase I, hydrogenase II catalyzed the H(2)-dependent reduction of NAD (K(m), 128 microM; k(cat)/K(m), 770 min(-1) mM(-1)). Ferredoxin from P. furiosus was not an efficient electron carrier for either enzyme. Both H(2) and NADPH served as electron donors for the reduction of elemental sulfur (S(0)) and polysulfide by hydrogenase I and hydrogenase II, and both enzymes preferentially reduce polysulfide to sulfide rather than protons to H(2) using NADPH as the electron donor. At least two [4Fe-4S] and one [2Fe-2S] cluster were detected in hydrogenase II by electron paramagnetic resonance spectroscopy, but amino acid sequence analyses indicated a total of five [4Fe-4S] clusters (two in the beta subunit and three in the delta subunit) and one [2Fe-2S] cluster (in the gamma subunit), as well as two putative nucleotide-binding sites in the gamma subunit which are thought to bind FAD and NAD(P)(H). The amino acid sequences of the four subunits of hydrogenase II showed between 55 and 63% similarity to those of hydrogenase I. The two enzymes are present in the cytoplasm at approximately the same concentration. Hydrogenase II may become physiologically relevant at low S(0) concentrations since it has a higher affinity than hydrogenase I for both S(0) and polysulfide.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Ma K,Weiss R,Adams MWdoi
10.1128/jb.182.7.1864-1871.2000keywords:
subject
Has Abstractpub_date
2000-04-01 00:00:00pages
1864-71issue
7eissn
0021-9193issn
1098-5530journal_volume
182pub_type
杂志文章abstract::Auxotrophic mutants of the filamentous cyanobacterium Anabaena variabilis were isolated by a method in which, after mutagenesis and before penicllin enrichment, mutant and wild-type cells were separated by cavitation. Auxotrophs were identified by their inability to grow on minimal medium, and they were partially char...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.129.3.1556-1562.1977
更新日期:1977-03-01 00:00:00
abstract::Random Tn917 mutagenesis of Bacillus subtilis followed by selection of lipoic acid auxotrophs led to the isolation of the cysH gene. The gene was sequenced and found to encode a phosphoadenylylsulfate sulfotransferase with a molecular mass of 27 kDa. Expression of lacZ fused to the cysH promoter was repressed by cyste...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.179.3.976-981.1997
更新日期:1997-02-01 00:00:00
abstract::Transducing frequencies of phage P22 lysates prepared from Salmonella typhimurium exponential cultures in minimal and nutrient broth media were compared. The assumption is that cells grown in a minimal medium will have one replication fork per replication unit, but cells in nutrient broth will have multiple replicatio...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.115.1.168-176.1973
更新日期:1973-07-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.123.3.1055-1067.1975
更新日期:1975-09-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.175.3.902-904.1993
更新日期:1993-02-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.99.3.842-850.1969
更新日期:1969-09-01 00:00:00
abstract::We present the complete genome sequence of Yersinia pestis KIM, the etiologic agent of bubonic and pneumonic plague. The strain KIM, biovar Mediaevalis, is associated with the second pandemic, including the Black Death. The 4.6-Mb genome encodes 4,198 open reading frames (ORFs). The origin, terminus, and most genes en...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.184.16.4601-4611.2002
更新日期:2002-08-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.113.1.252-262.1973
更新日期:1973-01-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.01077-08
更新日期:2009-02-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.01476-12
更新日期:2013-01-01 00:00:00
abstract::Polyribosomes detected in extracts of spores harvested with water also were found in extracts prepared from spores harvested with nonaqueous fluids. ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.122.3.1394-1395.1975
更新日期:1975-06-01 00:00:00
abstract::The activity of the ATP:GTP 3'-pyrophosphotransferase (guanosine pentaphosphate synthetase I [GPSI]) from Streptomyces antibioticus is stimulated maximally by methanol at 20% (vol/vol) in assay mixtures. Although the enzyme is not activated by ribosomes, its activity is stimulated by tRNA (uncharged or charged) and by...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.176.5.1482-1487.1994
更新日期:1994-03-01 00:00:00
abstract::Abram, Dinah (Purdue University, Lafayette, Ind.), A. E. Vatter, and Henry Koffler. Attachment and structural features of flagella of certain bacilli. J. Bacteriol. 91:2045-2068. 1966.-The attachment of flagella to cells of various mesophilic and thermophilic strains of Bacillus was studied electron microscopically. S...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.91.5.2045-2068.1966
更新日期:1966-05-01 00:00:00
abstract::The upper operon of the TOL plasmid pWW0 of Pseudomonas putida encodes a set of enzymes which transform toluene and xylenes to benzoate and toluates. The genetic organization of the operon was characterized by cloning of the upper operon genes into an expression vector and identification of their products in Escherich...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.171.9.5048-5055.1989
更新日期:1989-09-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:2008-12-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.01215-12
更新日期:2012-11-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.137.1.397-408.1979
更新日期:1979-01-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.145.1.145-155.1981
更新日期:1981-01-01 00:00:00
abstract::As part of our effort to uncover the molecular basis for the phenotypic variation among clinical Mycobacterium tuberculosis isolates, we have previously reported that isolates belonging to the W/Beijing lineage constitutively overexpress the DosR-regulated transcriptional program. While generating dosR knockouts in tw...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00536-10
更新日期:2010-09-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.110.1.41-46.1972
更新日期:1972-04-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:2011-10-01 00:00:00
abstract::Cystic fibrosis (CF) patients are highly susceptible to chronic pulmonary disease caused by mucoid Pseudomonas aeruginosa strains that overproduce the exopolysaccharide alginate. We showed here that a mutation in zwf, encoding glucose-6-phosphate dehydrogenase (G6PDH), leads to a approximately 90% reduction in alginat...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.187.22.7561-7568.2005
更新日期:2005-11-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.171.2.1219-1222.1989
更新日期:1989-02-01 00:00:00
abstract::Small subunit rDNA sequences were determined for 20 species of the genera Acetogenium, Clostridium, Thermoanaerobacter, Thermoanaerobacterium, Thermoanaerobium, and Thermobacteroides, 3 non-validly described species, and 5 isolates of anaerobic thermophilic bacteria, providing a basis for a phylogenetic analysis of th...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.175.15.4772-4779.1993
更新日期:1993-08-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.181.9.2872-2877.1999
更新日期:1999-05-01 00:00:00
abstract::Graber, C. D. (Baylor University College of Medicine, Houston, Tex.), R. M. O'Neal, and E. R. Rabin. Effect of high fat diets on intestinal microflora and serum cholesterol in rats. J. Bacteriol. 89:47-51. 1965.-Differential bacterial counts of feces and total plasma cholesterol determinations were performed on 60 Wis...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.89.1.47-51.1965
更新日期:1965-01-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.169.12.5546-5555.1987
更新日期:1987-12-01 00:00:00
abstract::To circumvent the permeability barrier of its outer membrane, Pseudomonas aeruginosa has evolved a series of specific porins. These channels have binding sites for related classes of molecules that facilitate uptake under nutrient-limited conditions. Here, we report on the identification of a 19-member family of porin...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.188.1.45-54.2006
更新日期:2006-01-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.95.1.231-238.1968
更新日期:1968-01-01 00:00:00
abstract::Chloromuconate cycloisomerases of bacteria utilizing chloroaromatic compounds are known to convert 3-chloro-cis,cis-muconate to cis-dienelactone (cis-4-carboxymethylenebut-2-en-4-olide), while usual muconate cycloisomerases transform the same substrate to the bacteriotoxic protoanemonin. Formation of protoanemonin req...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.183.15.4551-4561.2001
更新日期:2001-08-01 00:00:00