Kimitoshi Sakamoto

Ubiquinone (UQ) is a lipophilic electron carrier that functions in the respiratory and photosynthetic electron transfer chains of proteobacteria and eukaryotes. Bacterial UQ biosynthesis is well studied in the gammaproteobacterium Escherichia coli , in which most bacterial UQ‐biosynthetic enzymes have been identified. However, these enzymes are not always conserved among UQ‐containing bacteria. In particular, the alphaproteobacterial UQ biosynthesis pathways contain many uncharacterized steps with unknown features. In this work, we identified in the alphaproteobacterium Rhodobacter capsulatus a new decarboxylative hydroxylase and named it UbiN. Remarkably, the UbiN sequence is more similar to a salicylate hydroxylase than the conventional flavin‐containing UQ‐biosynthetic monooxygenases. Under aerobic conditions, R. capsulatus Δ ubiN mutant cells accumulate 3‐decaprenylphenol, which is a UQ‐biosynthetic intermediate. In addition, 3‐decaprenyl‐4‐hydroxybenzoic acid, which is the substrate of UQ‐biosynthetic decarboxylase UbiD, also accumulates in Δ ubiN cells under aerobic conditions. Considering that the R. capsulatus Δ ubiD‐X double mutant strain (UbiX produces a prenylated FMN required for UbiD) grows as a wild‐type strain under aerobic conditions, these results indicate that UbiN catalyzes the aerobic decarboxylative hydroxylation of 3‐decaprenyl‐4‐hydroxybenzoic acid. This is the first example of the involvement of decarboxylative hydroxylation in ubiquinone biosynthesis. This finding suggests that the C1 hydroxylation reaction is, at least in R. capsulatus , the first step among the three hydroxylation steps involved in UQ biosynthesis. Although the C5 hydroxylation reaction is often considered to be the first hydroxylation step in bacterial UQ biosynthesis, it appears that the R. capsulatus pathway is more similar to that found in mammalians.