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The enzymatic system thiosulfate: Cytochrome c oxidoreductase from photolithoautotrophically grown Rhodopseudomonas palustris

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Abstract

Rhodopseudomonas palustris cells, characterized by a lamellar type intracytoplasmic chromatophore membrane system after phototrophic growth, yielded a crude supernatant cell-free fraction (S-144) after ultracentrifugation which retained the contents of both the cell compartments. After thiosulfate-dependent growth, a protein system was isolated from S-144 which catalyzed the thiosulfate-linked reduction of an endogenous c-type cytochrome. — The colorless oxidoreductase protein, after purification to homogeneity, revealed a molecular weight of 93,000 and, after SDS treatment, a particle weight of 48,000. It was focused at an average pI of 5.45. Apparent K m values for several substrates were in the μM range. The electron acceptor for thiosulfate oxidation was found to be a cytochrome c from S-144. The homogeneous acceptor protein, at liquid nitrogen temperature, exhibited absorption maxima at 549.0, 518.5 and 418.0 nm, and shoulders at 525.5, 512.0 and 508.0 nm. Its molecular weight was found to be 17,000 (gel filtration) and 16,000 (SDS gel electrophoresis). It was characterized by a pI of 10.0. Its midpoint redox potential of E m,7.0=+228 mV was determined by redox titrations and the value of +205 mV by spectrophotometric calculations.

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Abbreviations

BSA:

bovine serum albumin

HiPIP:

high potential nonheme iron protein

IEF:

isoclectric focusing

SDS:

dodecylsulfate, sodium salt

Temed:

N,N,N′,N′-tetramethylethylenediamine

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Correspondence to Karl Knobloch.

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Schleifer, G., Schmitt, W. & Knobloch, K. The enzymatic system thiosulfate: Cytochrome c oxidoreductase from photolithoautotrophically grown Rhodopseudomonas palustris . Arch. Microbiol. 130, 328–333 (1981). https://doi.org/10.1007/BF00425950

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Key words

  • Photosynthesis
  • Rhodopseudomonas palustris
  • Thiosulfate
  • Cytochrome c
  • Oxidoreductase