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A crtA ‐related gene from Flavobacterium P99‐3 encodes a novel carotenoid 2‐hydroxylase involved in myxol biosynthesis
Author(s) -
Rählert Nina,
Fraser Paul D.,
Sandmann Gerhard
Publication year - 2009
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2009.04.025
Subject(s) - biosynthesis , complementation , escherichia coli , gene , biochemistry , flavobacterium , carotenoid , metabolic engineering , biology , structural gene , chemistry , genetics , bacteria , mutant , pseudomonas
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 .