2002
DOI: 10.1128/jb.184.23.6690-6699.2002
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Evolution of the C 30 Carotenoid Synthase CrtM for Function in a C 40 Pathway

Abstract: The C 30 carotene synthase CrtM from Staphylococcus aureus and the C 40 carotene synthase CrtB from Erwinia uredovora were swapped into their respective foreign C 40 and C 30 biosynthetic pathways (heterologously expressed in Escherichia coli) and evaluated for function. Each displayed negligible ability to synthesize the natural carotenoid product of the other. After one round of mutagenesis and screening, we isolated 116 variants of CrtM able to synthesize C 40 carotenoids. In contrast, we failed to find a s… Show more

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Cited by 73 publications
(70 citation statements)
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“…Green residues are involved in the first half-reaction (formation of PSPP), while blue residues form the pocket of the second half-reaction in which PSPP is rearranged. Red residues correspond by sequence alignment to F26 and W38 of CrtM, which were found to control the product size of that enzyme (213,214). Shown in yellow are the residues that align with mutations in CrtM that improve both its C 30 and C 40 carotenoid synthase activities.…”
Section: Directed Evolution Of Key Carotenoid Biosynthetic Enzymesmentioning
confidence: 99%
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“…Green residues are involved in the first half-reaction (formation of PSPP), while blue residues form the pocket of the second half-reaction in which PSPP is rearranged. Red residues correspond by sequence alignment to F26 and W38 of CrtM, which were found to control the product size of that enzyme (213,214). Shown in yellow are the residues that align with mutations in CrtM that improve both its C 30 and C 40 carotenoid synthase activities.…”
Section: Directed Evolution Of Key Carotenoid Biosynthetic Enzymesmentioning
confidence: 99%
“…Carotenoid synthases are quite specific with respect to product size, which reflects a strong preference for prenyl diphosphate substrates of a particular length. The C 40 carotenoid synthase CrtB does not detectably accept C 15 PP as a substrate, while the C 30 synthase CrtM accepts two molecules of C 20 PP to form C 40 carotenoids much less efficiently than it accepts two molecules of C 15 PP to form C 30 carotenoids (163,214). Even though various longer and shorter isoprenyl diphosphates are made by different organisms (146), no known carotenoids are derived from them.…”
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confidence: 99%
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