Bacillus schlegelii, a New Species of Thermophilic, Facultatively Chemolithoautotrophic Bacterium Oxidizing Molecular Hydrogen

Schenk, Anita; Aragno, Michel

doi:10.1099/00221287-115-2-333

Cited by 78 publications

(45 citation statements)

References 17 publications

(15 reference statements)

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“…In this work, we report the purification and some characterization of the hydrogenase from B. schlege/ii grown under heterotrophic conditions. The hydrogenase is constitutive in B. schlegelii 10 ) and the purified hydrogenase in this work showed the same properties as that reported in Ref. 11. B. schlege/ii (ATCC 43741) was grown heterotrophically with sodium pyruvate as a carbon source at 70°C as reported previously.l0.12) Cells were harvested by centrifugation and then washed with 50mM Tris-HC} buffer, pH 8.0.…”

supporting

confidence: 77%

Characterization and Thermostability of a Membrane-bound Hydrogenase from a Thermophilic Hydrogen Oxidizing Bacterium,Bacillus schlegelii

Aono

Kamachi

Okura

1993

Bioscience, Biotechnology, and Biochemistry

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Hydrogen oxidizing bacteria are aerobic chemolithotrophic bacteria containing hydrogenase as a key enzyme for hydrogen metabolism. 1) Two types of hydrogenase have been purified from hydrogen oxidizing bacteria, a soluble hydrogenase that reduce NAD with hydrogen 2 . 3 ) and a membrane-bound hydrogenase that does not react with NAD. 4 -8 ) The majority of hydrogen oxidizing bacteria contain merely a single membrane-bound hydrogenase.1.9) ,Bacillus schlegelii is a thermophilic (optimal growth temperature is 70°C) and facultatively chemolithotrophic bacterium and contains membrane-bound hydrogenase. 10. B. schlege/ii (ATCC 43741) was grown heterotrophically with sodium pyruvate as a carbon source at 70°C as reported previously.l0.12) Cells were harvested by centrifugation and then washed with 50mM Tris-HC} buffer, pH 8.0. The cell pellets were frozen in liquid nitrogen and stored at -80°C until use. The purification was done at room temperature under aerobic conditions. The frozen cells were resuspended in 50 mM Tris-HCI buffer, pH 8.0, containing deoxyribonuclease I (lOmg/ml buffer) and 4 mM MgCI 2 . After disruption of cells by sonication (Power; 60 W, treatment time; 1 min per g of wet cells), cell debris and unbroken cells were removed by centrifugation at 12,000 x g for 20 min. The membrane fraction was prepared by centrifugation at 160,000 x g for 60 min. The membrane fraction was resuspended (about 80 mg of the membrane per ml of buffer) and stirred gently at, room temperature for 60min in 50mM Tris-HCI buffer, pH 8.0, containing 0.1 % (wt/vol) Brij 58 to solubilize hydrogenase.When the hydrogenase was solubilized with the combination of 0.5% Triton X-IOO, 0.1 % sodium deoxycholate, and 5mM EDTA, 44.5% of hydrogenase activity was solubilized. 11) In this work, about 30% of hydrogenase activity was solubilized from the membrane as shown in Table. Although the efficiency of the solubilization by Brij 58 was slightly lower, itwas chosen because of the lack of an absorption band at 280 nm. The elution profile was monitored by the absorbance at 280 nm during the following column chromatography. Triton X-100 has so intense an absorption band at 280 nm that it interferes with monitoring the elution profile at 280 nm, but not Brij-58.The suspension was centrifuged at 160,000 x g for 60 min after the solubilization. The ,supernatant obtained was put on a column (2.5 x 30 cm) of DEAE-SepharoseFast Flow previously equilibrated with 50mM Tris-HCI buffer, pH 8.0, containing 0.02% Brij 58. The column was washed with 450 ml of the equilibration * To whom all correspondence should be addressed.buffer and then the adsorbed proteins were eluted with a 600-ml linear gradient from zero to 0.5 M NaCI at .3 mljmin. The hydrogenase started to elute when 0.24 M NaCI was put on the column.The hydrogenase-containing fractions were combined and concentrated to about 4rill by ultrafiltration (PM-30; Amicon Corp.). The sample was put on a column (1.5 x 90cm) ofSephacryl S-200 HR previously equilibrated with 50mM Tris-HCI buffer, pH 8....

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confidence: 77%

Characterization and Thermostability of a Membrane-bound Hydrogenase from a Thermophilic Hydrogen Oxidizing Bacterium,Bacillus schlegelii

Aono

Kamachi

Okura

1993

Bioscience, Biotechnology, and Biochemistry

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“…Cells were Gram-positive, spore-forming, oxidase-positive, long rods. The results on the cell morphology and other details are given in the species description and by Schenk and Aragno (1979).…”

Section: Cip 106933 T )mentioning

confidence: 99%

“…Bacillus schlegelii was proposed by Schenk and Aragno (1979) as a novel species of thermophilic, facultatively anaerobic, chemolithotrophic bacteria that are able to oxidize hydrogen. This study was a consequence of the initial study of Aragno (1978), who isolated several strains of spore-forming thermophilic hydrogen bacteria from the superficial layer of the sediment of a little eutrophic lake.…”

Section: Cip 106933 T )mentioning

confidence: 99%

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Transfer of Bacillus schlegelii to a novel genus and proposal of Hydrogenibacillus schlegelii gen. nov., comb. nov.

Kämpfer

Glaeser

Busse³

2013

International Journal of Systematic and Evolutionary Microbiology

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“…This strain, TK-6, cannot utilize organic compounds as the sole source of carbon and energy for growth; it is an obligately autotrophic hydrogen bacterium. This is unique, since all the aerobic hydrogen-oxidizing bacteria previously described including the moderate thermophiles 2 -4 ) and the extreme thermophile, 5,6) were facultative autotrophs which could grow heterotrophically utilizing organic compounds. 7 ) Obligate autotrophy was known S ) in the thiobacilli, the nitrifying bacteria and the blue-green algae but it was thought that there were no obligately autotrophic hydrogenoxidizing bacteria except Methanobacterium thermoautotrophicum, 9) an anaerobic, extremely thermophilic methanogenic bacterium.…”

mentioning

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The Deficient Carbohydrate Metabolic Pathways and the Incomplete Tricarboxylic Acid Cycle in an Obligately Autotrophic Hydrogen-oxidizing Bacterium

Shiba

Kawasumi

Igarashi

et al. 1982

Agricultural and Biological Chemistry

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Bacillus schlegelii, a New Species of Thermophilic, Facultatively Chemolithoautotrophic Bacterium Oxidizing Molecular Hydrogen

Cited by 78 publications

References 17 publications

Characterization and Thermostability of a Membrane-bound Hydrogenase from a Thermophilic Hydrogen Oxidizing Bacterium,Bacillus schlegelii

Characterization and Thermostability of a Membrane-bound Hydrogenase from a Thermophilic Hydrogen Oxidizing Bacterium,Bacillus schlegelii

Transfer of Bacillus schlegelii to a novel genus and proposal of Hydrogenibacillus schlegelii gen. nov., comb. nov.

The Deficient Carbohydrate Metabolic Pathways and the Incomplete Tricarboxylic Acid Cycle in an Obligately Autotrophic Hydrogen-oxidizing Bacterium

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