Purification and characterization of pyruvate:ferredoxin oxidoreductase from hydrogenobacter thermophilus TK-6

Ki Suk Yoon, Masaharu Ishii, Tohru Kodama, Yasuo Igarashi

研究成果: ジャーナルへの寄稿記事

31 引用 (Scopus)

抄録

Pyruvate:ferredoxin oxidoreductase was purified to electrophoretic homogeneity from an aerobic, thermophilic, obligately chemolithoautotrophic, hydrogenoxidizing bacterium, Hydrogenobacter thermophilus TK-6, by precipitation with ammonium sulfate and fractionation by DEAE-Sepharose CL-GB, polyacrylate-quaternary amine, hydroxyapatite, and Superdex-200 chromatography. The native enzyme had a molecular mass of 135 kDa and was composed of four different subunits with apparent molecular masses of 46, 31.5, 29, and 24.5 kDa, respectively, indicating that the enzyme has an αβγδ-structure. The activity was detected with pyruvate, coenzyme A, and one of the following electron accepters in substrate amounts: ferredoxin isolated from H. thermophilus, FAD, FMN, triphenyltetrazolium chloride, or methyl viologen. NAD, NADP, ana ferredoxins from Chlorella spp. and Clostridium pasteurianum were ineffective as the electron acceptor. The temperature optimum for pyruvate oxidation was approximately 80°C. The pH optimum was 7.6-7.8. The apparent K(m) values for pyruvate and coenzyme A at 70°C were 3.45 mM and 54 μM, respectively. The enzyme was extremely thermostable under anoxic conditions; the time for a 50% loss of activity (t(50%)) at 70°C was approximately 8 h.

元の言語英語
ページ(範囲)275-279
ページ数5
ジャーナルArchives of Microbiology
167
発行部数5
DOI
出版物ステータス出版済み - 4 28 1997
外部発表Yes

Fingerprint

Pyruvate Synthase
Pyruvic Acid
Purification
Ferredoxins
Molecular mass
Coenzyme A
Enzymes
Electrons
Chlorella
Flavin Mononucleotide
Clostridium
Flavin-Adenine Dinucleotide
Paraquat
Ammonium Sulfate
Durapatite
Fractionation
Chromatography
Polyacrylates
NADP
NAD

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Biochemistry
  • Molecular Biology
  • Genetics

これを引用

Purification and characterization of pyruvate:ferredoxin oxidoreductase from hydrogenobacter thermophilus TK-6. / Yoon, Ki Suk; Ishii, Masaharu; Kodama, Tohru; Igarashi, Yasuo.

:: Archives of Microbiology, 巻 167, 番号 5, 28.04.1997, p. 275-279.

研究成果: ジャーナルへの寄稿記事

Yoon, Ki Suk ; Ishii, Masaharu ; Kodama, Tohru ; Igarashi, Yasuo. / Purification and characterization of pyruvate:ferredoxin oxidoreductase from hydrogenobacter thermophilus TK-6. :: Archives of Microbiology. 1997 ; 巻 167, 番号 5. pp. 275-279.
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