Functional analysis of γ-carotene ketolase involved in the carotenoid biosynthesis of Deinococcus radiodurans

Deinococcus radiodurans strain R1 synthesizes a unique ketocarotenoid product named deinoxanthin. The detailed steps involved in the biosynthesis of deinoxanthin remain unresolved. A carotene ketolase homologue encoded by dr0093 was inactivated by gene mutation to verify its function in the native host D. radiodurans . Analysis of the carotenoids in the resultant mutant R1ΔcrtO demonstrated that dr0093 encodes γ-carotene ketolase (CrtO) catalysing the introduction of one keto group into the C-4 position of γ-carotene derivatives to form ketolated carotenoids. The mutant R1ΔcrtO became more sensitive to H₂O₂ treatment than the wild-type strain R1, indicating that the C-4 keto group is important for the antioxidant activity of carotenoids in D. radiodurans . Carotenoid extracts from mutant R1ΔcrtO exhibited lower 2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity than those from the wild-type strain R1. The enhanced antioxidant ability of ketocarotenoids in D. radiodurans might be attributed to its extended conjugated double bonds and relative stability by the C-4 keto group substitution.