The oxygen that we breathe is produced by photosystem II of cyanobacteria and plants. The catalytic centre, a Mn4Ca cluster, accumulates four oxidizing equivalents before oxygen is formed, seemingly in a single reaction step1,2,3,4,5,6,7,8 2H2O⇌O2 + 4H+ + 4e-. The energy and cycling of this reaction derives solely from light. No intermediate oxidation product of water has been detected so far. Here, we shifted the equilibrium of the terminal reaction backward by increasing the oxygen pressure and monitoring (by absorption transients in the near-ultraviolet spectrum) the electron transfer from bound water into the catalytic centre. A tenfold increase of ambient oxygen pressure (2.3 bar) half-suppressed the full progression to oxygen. The remaining electron transfer at saturating pressure (30 bar) was compatible with the formation of a stabilized intermediate. The abstraction of four electrons from water was probably split into at least two electron transfers: mildly endergonic from the centre's highest oxidation state to an intermediate, and exergonic from the intermediate to oxygen. There is little leeway for photosynthetic organisms to push the atmospheric oxygen concentration much above the present level.
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We thank H. Kenneweg for technical assistance, H. Heine for advice on the construction of the pressure cell, R. Debus for cooperation on Synechocystis, and R. Ahlbrink and A. Mulkidjanian for discussions and help. This work was financially supported by the Deutsche Forschungsgemeinschaft, the Fonds der Chemischen Industrie and the Land Niedersachsen.
The authors declare that they have no competing financial interests.
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Clausen, J., Junge, W. Detection of an intermediate of photosynthetic water oxidation. Nature 430, 480–483 (2004). https://doi.org/10.1038/nature02676
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