title: Additional diterpenes from Physcomitrella patens synthesized by copalyl diphosphate/kaurene synthase (PpCPS/KS).
abstract: The bifunctional diterpene synthase, copalyl diphosphate/kaurene synthase from the moss Physcomitrella patens (PpCPS/KS), catalyses the formation of at least four diterpenes, including ent-beyerene, ent-sandaracopimaradiene, ent-kaur-16-ene, and 16-hydroxy-ent-kaurene. The enzymatic activity has been confirmed through generation of a targeted PpCPS/KS knock-out mutant in P. patens via homologous recombination, through transient expression of PpCPS/KS in Nicotiana benthamiana, and expression of PpCPS/KS in E. coli. GC-MS analysis of the knock-out mutant shows that it lacks the diterpenoids, supporting that all are products of PpCPS/KS as observed in N. benthamiana and E. coli. These results provide additional knowledge of the mechanism of this bifunctional diterpene synthase, and are in line with proposed reaction mechanisms in kaurene biosynthesis.
authors.affiliation: Plant Biochemistry Laboratory, Copenhagen Plant Science Centre, Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, DK-1871 Frederiksberg, Denmark.
authors.forename: Xin
authors.initials: X
authors.lastname: Zhan
authors.affiliation: Plant Biochemistry Laboratory, Copenhagen Plant Science Centre, Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, DK-1871 Frederiksberg, Denmark.
authors.forename: Søren Spanner
authors.initials: SS
authors.lastname: Bach
authors.affiliation: Plant Biochemistry Laboratory, Copenhagen Plant Science Centre, Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, DK-1871 Frederiksberg, Denmark.
authors.forename: Nikolaj Lervad
authors.initials: NL
authors.lastname: Hansen
authors.affiliation: Plant Biochemistry Laboratory, Copenhagen Plant Science Centre, Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, DK-1871 Frederiksberg, Denmark.
authors.forename: Christina
authors.initials: C
authors.lastname: Lunde
authors.affiliation: Plant Biochemistry Laboratory, Copenhagen Plant Science Centre, Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, DK-1871 Frederiksberg, Denmark. Electronic address: hts@plen.ku.dk.
authors.forename: Henrik Toft
authors.initials: HT
authors.lastname: Simonsen
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000378:metabolism*; D019068:Bryopsida / Q000378:metabolism*; D004224:Diterpenes / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D010755:Organophosphates / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism*
chemical_list: D004224:Diterpenes; D010755:Organophosphates; D010940:Plant Proteins; C408747:copalyl diphosphate; D019883:Alkyl and Aryl Transferases; C090963:ent-kaurene synthetase A
keywords: Copalyl diphosphate/kaurene synthase; Diterpene; Ent-beyerene; Ent-sandaracopimaradiene; Physcomitrella patens