title: Total biosynthesis of the cyclic AMP booster forskolin fromColeus forskohlii.
abstract: Forskolin is a unique structurally complex labdane-type diterpenoid used in the treatment of glaucoma and heart failure based on its activity as a cyclic AMP booster. Commercial production of forskolin relies exclusively on extraction from its only known natural source, the plantColeus forskohlii, in which forskolin accumulates in the root cork. Here, we report the discovery of five cytochrome P450s and two acetyltransferases which catalyze a cascade of reactions converting the forskolin precursor 13R-manoyl oxide into forskolin and a diverse array of additional labdane-type diterpenoids. A minimal set of three P450s in combination with a single acetyl transferase was identified that catalyzes the conversion of 13R-manoyl oxide into forskolin as demonstrated by transient expression inNicotiana benthamiana. The entire pathway for forskolin production from glucose encompassing expression of nine genes was stably integrated intoSaccharomyces cerevisiaeand afforded forskolin titers of 40 mg/L.
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark.|Center for Synthetic Biology "bioSYNergy", Copenhagen, Denmark.
authors.forename: Irini
authors.identifier: 0000-0002-7526-2334
authors.initials: I
authors.lastname: Pateraki
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark.|Center for Synthetic Biology "bioSYNergy", Copenhagen, Denmark.
authors.forename: Johan
authors.initials: J
authors.lastname: Andersen-Ranberg
authors.affiliation: Evolva A/S, Copenhagen, Denmark.
authors.forename: Niels Bjerg
authors.initials: NB
authors.lastname: Jensen
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Sileshi Gizachew
authors.initials: SG
authors.lastname: Wubshet
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark.|Center for Synthetic Biology "bioSYNergy", Copenhagen, Denmark.
authors.forename: Allison Maree
authors.identifier: 0000-0002-2732-5185
authors.initials: AM
authors.lastname: Heskes
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Victor
authors.initials: V
authors.lastname: Forman
authors.affiliation: Science for Life Laboratory, KTH - Royal Institute of Technology, Stockholm, Sweden.
authors.forename: Björn
authors.initials: B
authors.lastname: Hallström
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark.|Center for Synthetic Biology "bioSYNergy", Copenhagen, Denmark.
authors.forename: Britta
authors.initials: B
authors.lastname: Hamberger
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark.|Center for Synthetic Biology "bioSYNergy", Copenhagen, Denmark.
authors.forename: Mohammed Saddik
authors.initials: MS
authors.lastname: Motawia
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark.|Center for Synthetic Biology "bioSYNergy", Copenhagen, Denmark.
authors.forename: Carl Erik
authors.initials: CE
authors.lastname: Olsen
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Dan
authors.initials: D
authors.lastname: Staerk
authors.affiliation: Evolva A/S, Copenhagen, Denmark.
authors.forename: Jørgen
authors.initials: J
authors.lastname: Hansen
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark.|Center for Synthetic Biology "bioSYNergy", Copenhagen, Denmark.
authors.forename: Birger Lindberg
authors.initials: BL
authors.lastname: Møller
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark.|Center for Synthetic Biology "bioSYNergy", Copenhagen, Denmark.
authors.forename: Björn
authors.identifier: 0000-0003-1249-1807
authors.initials: B
authors.lastname: Hamberger
mesh_terms: D053898:Biosynthetic Pathways / Q000235:genetics*; D001711:Biotransformation; D005576:Colforsin / Q000378:metabolism*; D004224:Diterpenes / Q000378:metabolism; D060847:Metabolic Engineering; D031346:Plectranthus / Q000235:genetics* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism
chemical_list: D004224:Diterpenes; C475535:manoyl oxide; D005576:Colforsin
keywords: Coleus forskohlii; biochemistry; diterpenoids biosynthesis; forskolin; pathway elucidation