title: Engineering biosynthesis of the anticancer alkaloid noscapine in yeast.
abstract: Noscapine is a potential anticancer drug isolated from the opium poppy Papaver somniferum, and genes encoding enzymes responsible for the synthesis of noscapine have been recently discovered to be clustered on the genome of P. somniferum. Here, we reconstitute the noscapine gene cluster in Saccharomyces cerevisiae to achieve the microbial production of noscapine and related pathway intermediates, complementing and extending previous in planta and in vitro investigations. Our work provides structural validation of the secoberberine intermediates and the description of the narcotoline-4'-O-methyltransferase, suggesting this activity is catalysed by a unique heterodimer. We also reconstitute a 14-step biosynthetic pathway of noscapine from the simple alkaloid norlaudanosoline by engineering a yeast strain expressing 16 heterologous plant enzymes, achieving reconstitution of a complex plant pathway in a microbial host. Other engineered yeasts produce previously inaccessible pathway intermediates and a novel derivative, thereby advancing protoberberine and noscapine related drug discovery.
authors.affiliation: Shriram Center, Department of Bioengineering, Stanford University, 443 Via Ortega, MC 4245, Stanford, California 94305, USA.
authors.forename: Yanran
authors.initials: Y
authors.lastname: Li
authors.affiliation: Shriram Center, Department of Bioengineering, Stanford University, 443 Via Ortega, MC 4245, Stanford, California 94305, USA.
authors.forename: Christina D
authors.initials: CD
authors.lastname: Smolke
mesh_terms: D000970:Antineoplastic Agents / Q000378:metabolism*; D001600:Berberine Alkaloids; D057005:Bioengineering / Q000379:methods*; D053898:Biosynthetic Pathways; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D055808:Drug Discovery / Q000379:methods; D008780:Methyltransferases / Q000378:metabolism; D005810:Multigene Family; D009665:Noscapine / Q000378:metabolism*; D010207:Papaver / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae; D013765:Tetrahydropapaveroline / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D001600:Berberine Alkaloids; D010940:Plant Proteins; C009090:protoberberine; D013765:Tetrahydropapaveroline; D009665:Noscapine; D003577:Cytochrome P-450 Enzyme System; D008780:Methyltransferases