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

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

研究成果: Contribution to journalArticle査読

32 被引用数 (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.

本文言語英語
ページ(範囲)275-279
ページ数5
ジャーナルArchives of Microbiology
167
5
DOI
出版ステータス出版済み - 1997
外部発表はい

All Science Journal Classification (ASJC) codes

  • 微生物学
  • 生化学
  • 分子生物学
  • 遺伝学

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