A crtA-related gene from Flavobacterium P99-3 encodes a novel carotenoid 2-hydroxylase involved in myxol biosynthesis |
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Authors: | Nina Rählert Gerhard Sandmann |
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Institution: | a Biosynthesis Group, Molecular Biosciences 213, Department of Biological Sciences, J.W. Goethe Universität, P.O. Box 111932, D-60054 Frankfurt, Germany b School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey TW20 OEX, UK |
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Abstract: | A genomic DNA fragment with carotenogenic genes involved in myxol biosynthesis (3′,4′-didehydro-1′,2′-dihydro-β,ψ-carotene-3,1′,2′-triol) was cloned from Flavobacterium P99-3. It contains a gene highly homologous to crtA from purple bacteria encoding there an acyclic carotenoid 2-ketolase. Since no ketolation step is involved in myxol biosynthesis, the function of crtA-OH from Flavobacterium was assigned by complementation in Escherichia coli engineered to synthesize demethylspheroidene and 1′-hydroxy-demethylspheroidene. Upon co-expression of crtA-OH, the formation of 2-hydroxy derivatives of both carotenoids assigns CrtA-OH as a novel carotenoid hydroxylase. The gene was used to re-construct myxol biosynthesis in E. coli successfully. Additionally, 1′,2′-dihydroxytorulene and 1,2,1′-trihydroxy-3,4,3′,4′-tetradehydrolycopene were obtained. Their generation demonstrates that a new class of 2-hydroxy carotenoids can now be pursued by genetic engineering in E. coli. |
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Keywords: | Carotenogenic gene cluster Carotenoid 2-hydroxylase 2-Hydroxy-demethylspheroidene 2 1&prime -Dihydroxy-demethylspheroidene Myxol crtA gene Flavobacterium P99-3 |
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