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Untersuchungen zum Cytochrom-Oxydase-System aus anaerob im Licht und aerob im Dunkeln gewachsenen Zellen von Rhodopseudomonas capsulata

Studies on the cytochrome oxidase system of ligh-anaerobically and dark-aerobically grown cells of Rhodopseudomonas capsulata

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Summary

The aerobic oxidase-system from dark-aerobically and light-anaerobically grown Rps. capsulata was investigated. The particulate fraction sedimented from ultrasonic extracts by 140,000 g-centrifugation, catalyzed the oxidation of NADH, succinate and reduced cytochrome c (horse heart). The oxidase activities of the particles from dark-aerobically grown cells were in the range of 0.10–0.38 μmoles O2/min x mg protein and were usually ten times as high as the oxidase activities from light-grown cells. The particles contain cytochromes of b-type and c-type. Cytochrome of a-type could be detected neither in the particles from light-grown nor in the particles from dark-grown cells. The highest values of the relation between cytochrome b and cytochrome c were found in the particles from darkaerobically grown cells. The cytochrome oxidase reacts with cytochrome c (horse heart), DCPIP and TMPD. The k m -values are 5×10-5 m, 1.5×10-4 m, or 2×10-4 m, respectively. The cytochrome oxidase exhibits a broad pH-optimum in the range of pH 8.5–9.5. Saturation of the oxidase with O2 is observed at a partial pressure of 15–20% O2. The oxidase is inhibited by KCN and NaN3 (half inhibition at 10-5 m), but not by CO. The particles from dark-aerobically and light-anaerobically grown cells catalyze phosphorylation of ADP in the dark coupled to the oxidation of succinate with maximum P/O-values of 0.3.

Zusammenfassung

Das aerobe Oxydase-System aus aerob im Dunkeln und anaerob im Licht gewachsenen Zellen von Rps. capsulata wurde untersucht. Die aus Ultraschallextrakten durch 140 000 g-Zentrifugation gewonnen Partikelfraktion katalysiert die Oxydation von NADH, Succinat und reduziertem Cytochrom c (aus Pferdeherz). Die Oxydase-Aktivitäten der Partikel aus aerob im Dunkeln gewachsenen Zellen variieren von 0,10–0,38 μmole O2/min · mg Protein und sind im Durchschnitt 10mal höher als die Oxydase-Aktivitäten der Partikel aus anaerob im Licht gewachsenen Zellen. Die Partikel enthalten Cytochrom vom b-Typ und c-Typ. Cytochrom a konnte weder in den Partikeln aus anaerob im Licht noch in den Partikeln aus aerob im Dunkeln gewachsenen Zellen nachgewiesen werden. In den Partikeln aus aerob gewachsenen Zellen ist das Verhältnis Cytochrom b:Cytochrom c größer als in den Partikeln aus anaerob im Licht gewachsenen Zellen. Die Cytochromoxydase reagiert mit Cytochrom c (aus Pferdeherz), DCPIP und TMPD. Die entsprechenden k m -Werte betragen 5 · 10-5 m, 1,5 · 10-4 m und 2 · 10-4 m. Die Cytochromoxydase hat ein breites pH-Optimum zwischen pH 8,5 und 9,5. Sättigung der Oxydase mit O2 tritt erst bei einem Partialdruck von 15 bis 20% O2 ein. Die Oxydase wird durch KCN und NaN3 (50% Hemmung bei 10-5 m), nicht aber durch CO gehemmt. Die Partikel aus aerob im Dunkeln und anaerob im Licht gewachsenen Zellen katalysieren eine mit der Succinat-Oxydation einhergehende Phosphorylierung von ADP mit P/O-Werten von maximal 0,3.

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Abbreviations

ADP:

Adenosindiphosphat

ATP:

Adenosintriphosphat

BChl:

Bacteriochlorophyll

CCCP:

Carbonylcyanid-m-chlorphenylhydrazon

DCPIP:

2,6-Dichlorphenolindophenol

DNP:

2,4-Dinitrophenol

NAD(P):

Nicotinamid-Adenin-Dinucleotid(phosphat)

NAD(P)H:

reduziertes NAD(P)

R. :

Rhodospirillum

Rps. :

Rhodopseudomonas

TMPD:

N-Tetramethyl-p-phenylendiamin

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Authors and Affiliations

  1. Institut für Mikrobiologie der Universität Göttingen, Göttingen, Deutschland

    J. -H. Klemme & H. G. Schlegel

  2. Department of Microbiology, Indiana University, Jordan Hall 438, 47401, Bloomington, Indiana, USA

    J. -H. Klemme

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  1. J. -H. Klemme
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  2. H. G. Schlegel
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Klemme, J.H., Schlegel, H.G. Untersuchungen zum Cytochrom-Oxydase-System aus anaerob im Licht und aerob im Dunkeln gewachsenen Zellen von Rhodopseudomonas capsulata . Arch. Mikrobiol. 68, 326–354 (1969). https://doi.org/10.1007/BF00408858

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