Affiliation:
1. Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125
Abstract
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.
Publisher
American Society for Microbiology
Subject
Molecular Biology,Microbiology
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