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Electron-transport chain and coupled oxidative phosphorylation in methanol-grown Paracoccus denitrificans

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Abstract

Methanol dehydrogenase of Paracoccus denitrificans was shown to be very similar to the enzyme of Pseudomonas sp, M. 27. The K m value for methanol with excess activator (ammonium ions) is 35 μM. The pH optimum for enzyme activity with 2,6-dichlorophe-nolindophenol as electronacceptor was at 9.0 A CO-binding type of cytochrome c was present only in cells grown with methanol as carbon and energy source.

It has been shown that methanol-oxidation involves electron-transport via cytochrome c and an a-type cytochrome to oxygen. Antimycin A did not inhibit this electron transport and 90% inhibition was obtained by 375 μM potassium cyanide. Electron transport from endogenous substrates is possible via cytochrome b and possibly cytochrome o to oxygen. Potassium cyanide inhibited 90% of the electron transport via this pathway at a concentration of 1.42 mM. Measurement of respiration-driven proton translocation proved that during oxidation of methanol to formaldehyde by oxygen one mole of adenosine triphosphate is synthesized in the site 3 region of the electron transport chain. The → H+/O value found confirmed the → H+/site ratio of 3–4 found in heterotrophic grown cells. During electron transport from endogenous substrates to oxygen there is a possible synthesis of 3 moles of adenosine triphosphate.

In heterotrophically grown cells electron transfer to oxygen follows almost only the branch of the respiratory chain containing cytochrome o. In methanol-grown cells the pathway via the a-type cytochrome seems more important.

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Abbreviations

DCPIP:

2,6-dichlorophenolindophenol

PMS:

phenazine methosulphate

EPR:

electron paramagnetic resonance

S.D.:

standard deviation

ATP:

adenosine triphosphate

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Authors and Affiliations

  1. Department of Microbiology, Biological Laboratory, Free University, De Boelelaan 1087, Amsterdam, The Netherlands

    H. W. Van Verseveld & A. H. Stouthamer

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  1. H. W. Van Verseveld
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  2. A. H. Stouthamer
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Van Verseveld, H.W., Stouthamer, A.H. Electron-transport chain and coupled oxidative phosphorylation in methanol-grown Paracoccus denitrificans . Arch. Microbiol. 118, 13–20 (1978). https://doi.org/10.1007/BF00406068

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