It is demonstrated that cyanobacteria (both azotrophic and non-azotrophic) contain hemeboxidoreductases that can convert chlorite to chloride and molecular oxygen (incorrectly denominated chlorite ‘dismutase’, Cld). Beside the water-splitting manganese complex of photosystem II, this metalloenzyme is the second known enzyme that catalyses the formation of a covalent oxygen–oxygen bond. All cyanobacterial Clds have a truncated N-terminus and are dimeric (i.e. clade 2) proteins. As model protein, Cld fromCyanothecesp. PCC7425 (CCld) was recombinantly produced inEscherichia coliand shown to efficiently degrade chlorite with an activity optimum at pH 5.0 [kcat1144 ± 23.8 s−1,KM162 ± 10.0 μM, catalytic efficiency (7.1 ± 0.6) × 106 M−1 s−1]. The resting ferric high-spin axially symmetric heme enzyme has a standard reduction potential of the Fe(III)/Fe(II) couple of −126 ± 1.9 mV at pH 7.0. Cyanide mediates the formation of a low-spin complex withkon= (1.6 ± 0.1) × 105 M−1 s−1 andkoff= 1.4 ± 2.9 s−1 (KD˜ 8.6 μM). Both, thermal and chemical unfolding follows a non-two-state unfolding pathway with the first transition being related to the release of the prosthetic group. The obtained data are discussed with respect to known structure–function relationships of Clds. We ask for the physiological substrate and putative function of these O2-producing proteins in (nitrogen-fixing) cyanobacteria.
First comprehensive biochemical and biophysical characterization of a cyanobacterial chlorite dismutase (Cld). The homodimeric (clade 2) heme enzyme efficiently degrades chlorite to chloride and dioxygen. Alternative substrates are tested and obtained data are discussed with respect to the physiological function of Cld in cyanobacteria.