In vitro characterization of a carotenoid cleavage dioxygenase from Nostoc sp. PCC 7120 reveals a novel cleavage pattern, cytosolic localization and induction by highlight

Carotenoid oxygenases catalyse the cleavage of C-C double bonds forming apocarotenoids, a diverse group of compounds, including retinoids and the precursors of some phytohormones. Some apocarotenoids, like β-ionone (C₁₃), are ecologically important volatiles released by plants and cyanobacteria. In this work, we elucidated the activity of the N ostoc c arotenoid c leavage d ioxygenase (NosCCD, previously named NSC1) using synthetic and cyanobacterial substrates. NosCCD converted bicyclic and monocyclic xanthophylls, including myxoxanthophylls, glycosylated carotenoids that are essential for thylakoid and cell wall structure. The products identified revealed two different cleavage patterns. The first is observed with bicyclic xanthophylls and is identical with that of plant orthologues, while the second is novel and occurs upon cleavage of monocyclic substrates at the C9-C10 and C7'-C8' double bonds. These properties enable the enzyme to produce a plenitude of different C₁₀ and C₁₃ apocarotenoids. Expression analyses indicated a role of NosCCD in response to highlight stress. Western blot analyses of Nostoc cells revealed NosCCD as a soluble enzyme in the cytosol, which also accomodates NosCCD substrates. Incubation of the corresponding fraction with synthetic substrates revealed the activity of the native enzyme and confirmed its induction by highlight.