title: Rapid identification of enzyme variants for reengineered alkaloid biosynthesis in periwinkle.
abstract: Monoterpene indole alkaloids from Catharanthus roseus (Madagascar periwinkle), such as the anticancer agents vinblastine and vincristine, have important pharmacological activities. Metabolic engineering of alkaloid biosynthesis can provide an efficient and environmentally friendly route to analogs of these synthetically challenging and pharmaceutically valuable natural products. However, the narrow substrate scope of strictosidine synthase, the enzyme at the entry point of the pathway, limits a pathway engineering approach. We demonstrate that with a different expression system and screening method it is possible to rapidly identify strictosidine synthase variants that accept tryptamine analogs not turned over by the wild-type enzyme. The variants are used in stereoselective synthesis of beta-carboline analogs and are assessed for biosynthetic competence within the terpene indole alkaloid pathway. These results present an opportunity to explore metabolic engineering of "unnatural" product production in the plant periwinkle.
authors: Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors: Peter
authors: P
authors: Bernhardt
authors: Elizabeth
authors: E
authors: McCoy
authors: Sarah E
authors: SE
authors: O'Connor
mesh_terms: D001483:Base Sequence; D030008:Catharanthus / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003124:Colorimetry; D017931:DNA Primers; D026121:Indole Alkaloids / Q000378:metabolism*; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D016296:Mutagenesis
chemical_list: D017931:DNA Primers; D026121:Indole Alkaloids
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