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Archives of Microbiology

, Volume 163, Issue 1, pp 21–28 | Cite as

Pyruvate: ferredoxin oxidoreductase from the sulfate-reducing Archaeoglobus fulgidus: molecular composition, catalytic properties, and sequence alignments

  • Jasper Kunow
  • Dietmar Linder
  • Rudolf K. Thauer
Original Paper

Abstract

Archaeoglobus fulgidus is a hyperthermophilic sulfate-reducing archaeon. In this communication we describe the purification and properties of pyruvate: ferredoxin oxidoreductase from this organism. The catabolic enzyme was purified 250-fold to apparent homogeneity with a yield of 16%. The native enzyme had an apparent molecular mass of 120 kDa and was composed of four different subunits of apparent molecular masses of 45, 33, 25, and 13 kDa, indicating and α β γ δ structure. Per mol, the enzyme contained 0.8 mol thiamine pyrophosphate, 9 mol non-heme iron, and 8 mol acid-labile sulfur. FAD, FMN, lipoic acid, and copper were not found. The purified enzyme showed an apparent Km for coenzyme A of 0.02 mM, for pyruvate of 0.3 mM, and for clostridial ferredoxin of 0.01 mM, an apparent Vmax of 64 U/mg (at 65°C) with a pH optimum near 7.5 and an Arrhenius activation energy of 75 kJ/mol (between 30 and 70°C). The temperature optimum was above 90°C. At 90°C, the enzyme lost 50% activity within 60 min in the presence of 2 M KCl. The enzyme did not catalyze the oxidation of 2-oxoglutarate, indolepyruvate, phenylpyruvate, glyoxylate, and hydroxypyruvate. The N-terminal amino acid sequences of the four subunits were determined. The sequence of the α-subunit had similarities to the N-terminal amino acid sequence of the α-subunit of the heterotetrameric pyruvate: ferredoxin oxidoreductase from Pyrococcus furiosus and from Thermotoga maritima, and unexpectedly, to the N-terminal amino acid sequence of the homodimeric pyruvate: ferredoxin oxidoreductase from proteobacteria and from cyanobacteria. No sequence similarities were found, however, between the α-subunits of the enzyme from A. fulgidus and the heterodimeric pyruvate: ferredoxin oxidoreductase from Halobacterium halobium.

Key words

Archaea Archaeoglobus Dissimilatory sulfate reduction Hyperthermophiles Pyruvate: ferredoxin (flavodoxin) oxidoreductase 

Abbreviations

CoASH

Coenzyme A

F420

Coenzyme F420

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Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • Jasper Kunow
    • 1
  • Dietmar Linder
    • 2
  • Rudolf K. Thauer
    • 1
  1. 1.Laboratorium für Mikrobiologie des Fachbereichs Biologie der Philipps-Universität and Max-Planck-Institut für terrestrische MikrobiologieMarburgGermany
  2. 2.Biochemisches Institut, Fachbereich HumanmedizinJustus-Liebig-UniversitätGießenGermany

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