title: Metabolic Engineering of Synechocystis sp. PCC 6803 for Production of the Plant Diterpenoid Manoyl Oxide.
abstract: Forskolin is a high value diterpenoid with a broad range of pharmaceutical applications, naturally found in root bark of the plant Coleus forskohlii. Because of its complex molecular structure, chemical synthesis of forskolin is not commercially attractive. Hence, the labor and resource intensive extraction and purification from C. forskohlii plants remains the current source of the compound. We have engineered the unicellular cyanobacterium Synechocystis sp. PCC 6803 to produce the forskolin precursor 13R-manoyl oxide (13R-MO), paving the way for light driven biotechnological production of this high value compound. In the course of this work, a new series of integrative vectors for use in Synechocystis was developed and used to create stable lines expressing chromosomally integrated CfTPS2 and CfTPS3, the enzymes responsible for the formation of 13R-MO in C. forskohlii. The engineered strains yielded production titers of up to 0.24 mg g(-1) DCW 13R-MO. To increase the yield, 13R-MO producing strains were further engineered by introduction of selected enzymes from C. forskohlii, improving the titer to 0.45 mg g(-1) DCW. This work forms a basis for further development of production of complex plant diterpenoids in cyanobacteria.
authors.affiliation: Department of Chemistry-Ångström, Uppsala University , Box 523, SE-751 20 Uppsala, Sweden.
authors.forename: Elias
authors.initials: E
authors.lastname: Englund
authors.affiliation: Department of Plant and Environmental Sciences, Center for Synthetic Biology bioSYNergy, Faculty of Science, University of Copenhagen , Thorvaldsensvej 40, 1871 Frederiksberg C, Copenhagen, Denmark.
authors.forename: Johan
authors.initials: J
authors.lastname: Andersen-Ranberg
authors.affiliation: Department of Chemistry-Ångström, Uppsala University , Box 523, SE-751 20 Uppsala, Sweden.
authors.forename: Rui
authors.initials: R
authors.lastname: Miao
authors.affiliation: Department of Plant and Environmental Sciences, Center for Synthetic Biology bioSYNergy, Faculty of Science, University of Copenhagen , Thorvaldsensvej 40, 1871 Frederiksberg C, Copenhagen, Denmark.
authors.forename: Björn
authors.initials: B
authors.lastname: Hamberger
authors.affiliation: Department of Chemistry-Ångström, Uppsala University , Box 523, SE-751 20 Uppsala, Sweden.
authors.forename: Pia
authors.initials: P
authors.lastname: Lindberg
mesh_terms: D004224:Diterpenes / Q000378:metabolism*; D005964:Glucosyltransferases / Q000096:biosynthesis / Q000235:genetics; D060847:Metabolic Engineering; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics; D031346:Plectranthus / Q000201:enzymology / Q000235:genetics*; D046939:Synechocystis / Q000235:genetics / Q000378:metabolism
chemical_list: D004224:Diterpenes; D010940:Plant Proteins; C475535:manoyl oxide; D005964:Glucosyltransferases
keywords: MEP-pathway; Synechocystis; diterpenoid; forskolin; genetic tools; manoyl oxide