Triplet-driven chemical reactivity of β-carotene and its biological implications

The endoperoxides of β-carotene (βCar-EPOs) are regarded as main products of the chemical deactivation of ¹O₂ by β-carotene, one of the most important antioxidants, following a concerted singlet-singlet reaction. Here we challenge this view by showing that βCar-EPOs are formed in the absence of ¹O₂ in a non-concerted triplet-triplet reaction: ³O₂ + ³β-carotene → βCar-EPOs, in which ³β-carotene manifests a strong biradical character. Thus, the reactivity of β-carotene towards oxygen is governed by its excited triplet state. βCar-EPOs, while being stable in the dark, are photochemically labile, and are a rare example of nonaromatic endoperoxides that release ¹O₂, again not in a concerted reaction. Their light-induced breakdown triggers an avalanche of free radicals, which accounts for the pro-oxidant activity of β-carotene and the puzzling swap from its anti- to pro-oxidant features. Furthermore, we show that βCar-EPOs, and carotenoids in general, weakly sensitize ¹O₂. These findings underlie the key role of the triplet state in determining the chemical and photophysical features of β-carotene. They shake up the prevailing models of carotenoid photophysics, the anti-oxidant functioning of β-carotene, and the role of ¹O₂ in chemical signaling in biological photosynthetic systems. βCar-EPOs and their degradation products are not markers of ¹O₂ and oxidative stress but of the overproduction of extremely hazardous chlorophyll triplets in photosystems. Hence, the chemical signaling of overexcitation of the photosynthetic apparatus is based on a ³chlorophyll-³β-carotene relay, rather than on extremely short-lived ¹O₂.