Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark; Center for Synthetic Biology “bioSYNergy”, Copenhagen, Denmark
Johan Andersen-Ranberg
Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark; Center for Synthetic Biology “bioSYNergy”, Copenhagen, Denmark
Niels Bjerg Jensen
Evolva A/S, Copenhagen, Denmark
Sileshi Gizachew Wubshet
Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark; Center for Synthetic Biology “bioSYNergy”, Copenhagen, Denmark
Victor Forman
Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark
Björn Hallström
Science for Life Laboratory, KTH - Royal Institute of Technology, Stockholm, Sweden
Britta Hamberger
Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark; Center for Synthetic Biology “bioSYNergy”, Copenhagen, Denmark
Mohammed Saddik Motawia
Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark; Center for Synthetic Biology “bioSYNergy”, Copenhagen, Denmark
Carl Erik Olsen
Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark; Center for Synthetic Biology “bioSYNergy”, Copenhagen, Denmark
Dan Staerk
Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
Jørgen Hansen
Evolva A/S, Copenhagen, Denmark
Birger Lindberg Møller
Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark; Center for Synthetic Biology “bioSYNergy”, Copenhagen, Denmark
Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark; Center for Synthetic Biology “bioSYNergy”, Copenhagen, Denmark
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 plant Coleus 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 in Nicotiana benthamiana. The entire pathway for forskolin production from glucose encompassing expression of nine genes was stably integrated into Saccharomyces cerevisiae and afforded forskolin titers of 40 mg/L.
