The obligate respiratory supercomplex from Actinobacteria

Abstract Actinobacteria are closely linked to human life as industrial producers of bioactive molecules and as human pathogens. Respiratory cytochrome bcc complex and cytochrome aa 3 oxidase are key components of their aerobic energy metabolism. They form a supercomplex in the actinobacterial species Corynebacterium glutamicum . With comprehensive bioinformatics and phylogenetic analysis we show that genes for cyt bcc - aa 3 supercomplex are characteristic for Actinobacteria ( Actinobacteria and Acidimicrobiia , except the anaerobic orders Actinomycetales and Bifidobacteriales ). An obligatory supercomplex is likely, due to the lack of genes encoding alternative electron transfer partners such as mono-heme cyt c . Instead, subunit QcrC of bcc complex, here classified as short di-heme cyt c , will provide the exclusive electron transfer link between the complexes as in C. glutamicum . Purified to high homogeneity, the C. glutamicum bcc - aa 3 supercomplex contained all subunits and cofactors as analyzed by SDS-PAGE, BN-PAGE, absorption and EPR spectroscopy. Highly uniform supercomplex particles in electron microscopy analysis support a distinct structural composition. The supercomplex possesses a dimeric stoichiometry with a ratio of a -type, b -type and c -type hemes close to 1:1:1. Redox titrations revealed a low potential bcc complex (E m ISP  = + 160 mV, E m b L  = − 291 mV, E m b H  = − 163 mV, E m cc  = + 100 mV) fined-tuned for oxidation of menaquinol and a mixed potential aa 3 oxidase (E m CuA  = + 150 mV, E m a / a 3  = + 143/+317 mV) mediating between low and high redox potential to accomplish dioxygen reduction. The generated molecular model supports a stable assembled supercomplex with defined architecture which permits energetically efficient coupling of menaquinol oxidation and dioxygen reduction in one supramolecular entity.