PMID:14973014
| Citation |
Umeno, D and Arnold, FH (2004) Evolution of a pathway to novel long-chain carotenoids. J. Bacteriol. 186:1531-6 |
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| Abstract |
Using methods of laboratory evolution to force the C(30) carotenoid synthase CrtM to function as a C(40) synthase, followed by further mutagenesis at functionally important amino acid residues, we have discovered that synthase specificity is controlled at the second (rearrangement) step of the two-step reaction. We used this information to engineer CrtM variants that can synthesize previously unknown C(45) and C(50) carotenoid backbones (mono- and diisopentenylphytoenes) from the appropriate isoprenyldiphosphate precursors. With this ability to produce new backbones in Escherichia coli comes the potential to generate whole series of novel carotenoids by using carotenoid-modifying enzymes, including desaturases, cyclases, hydroxylases, and dioxygenases, from naturally occurring pathways. |
| Links | |
| Keywords |
Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Carotenoids/metabolism; Directed Molecular Evolution; Erwinia/enzymology; Erwinia/genetics; Escherichia coli/enzymology; Escherichia coli/genetics; Farnesyl-Diphosphate Farnesyltransferase/genetics; Farnesyl-Diphosphate Farnesyltransferase/metabolism; Genetic Engineering/methods; Models, Molecular; Mutagenesis |
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