TAILIEUCHUNG - Báo cáo khoa học: The first cytochrome P450 in ferns Evidence for its involvement in phytoecdysteroid biosynthesis in Polypodium vulgare

The fernPolypodium vulgareis a phytoecdysteroid (PE)-producing plant. Cultures ofP. vulgareprothalus produce PE, whereas prothalus-derived callus cultures do not. However, this callus line can transform topically applied ecdysone (E) to 20-hydroxyecdysone (20E), which is the last step in the biosynthetic pathway of the main plant PE. This hydroxylation is cata-lysed by a cytochrome P450 enzyme. | ềFEBS Journal The first cytochrome P450 in ferns Evidence for its involvement in phytoecdysteroid biosynthesis in Polypodium vulgare Daniel Canals Josep Irurre-Santilari and Josefina Casas Department of BiologicalOrganic Chemistry IIQAB-CSIC Barcelona Spain Keywords cytochrome P450 ferns phytoecdysteroids sterolpathways Correspondence J. Casas Department of BiologicalOrganic Chemistry IIQAB-CSIC Jordi Girona 18-26 08034 Barcelona Spain Fax 34 93 204 5904 Tel 34 93 400 6100 E-mail jcbqob@ Received 14 March 2005 revised 19 July 2005 accepted 3 August 2005 doi The fern Polypodium vulgare is a phytoecdysteroid PE -producing plant. Cultures of P. vulgare prothalus produce PE whereas prothalus-derived callus cultures do not. However this callus line can transform topically applied ecdysone E to 20-hydroxyecdysone 20E which is the last step in the biosynthetic pathway of the main plant PE. This hydroxylation is catalysed by a cytochrome P450 enzyme. E treatment of the callus line results in an increased amount of P450 showing a linear correspondence between the amount of P450 and in vivo E 20-hydroxylation activity estimated by measuring the bioconversion of E to 20E. This activity can be inhibited by molecules that bind to the P450-heme group. E shows a P450-substrate-binding spectrum with microsomes that overexpress the P450 protein. Finally a P450 protein was purified from E-treated calli this being the first P450 to be described in the pterydophyte group. Ecdysteroids are involved at every stage in the lifecycle of arthropods regulating molting metamorphosis development reproduction and many of the physiological biochemical processes associated with those events. In insects the prothoracic glands secrete ecdysone E which is subsequently transformed into the physiologically more active ecdysteroid 20-hydroxy-ecdysone 20E 1 . Moreover ecdysteroids are also present in plants phytoecdysteroids PE but currently no phylogenetic .

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