title: Characterization of a dynamic metabolon producing the defense compound dhurrin in sorghum.
abstract: Metabolic highways may be orchestrated by the assembly of sequential enzymes into protein complexes, or metabolons, to facilitate efficient channeling of intermediates and to prevent undesired metabolic cross-talk while maintaining metabolic flexibility. Here we report the isolation of the dynamic metabolon that catalyzes the formation of the cyanogenic glucoside dhurrin, a defense compound produced in sorghum plants. The metabolon was reconstituted in liposomes, which demonstrated the importance of membrane surface charge and the presence of the glucosyltransferase for metabolic channeling. We used in planta fluorescence lifetime imaging microscopy and fluorescence correlation spectroscopy to study functional and structural characteristics of the metabolon. Understanding the regulation of biosynthetic metabolons offers opportunities to optimize synthetic biology approaches for efficient production of high-value products in heterologous hosts.
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Science, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.|bioSYNergy, Center for Synthetic Biology, DK-1871 Frederiksberg C, Denmark.|VILLUM Research Center for Plant Plasticity, DK-1871 Frederiksberg C, Denmark.|Copenhagen Plant Science Center, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.|Feedstocks Division, Joint BioEnergy Institute, Emeryville, CA 94608, USA.
authors.forename: Tomas
authors.initials: T
authors.lastname: Laursen
authors.affiliation: bioSYNergy, Center for Synthetic Biology, DK-1871 Frederiksberg C, Denmark.|VILLUM Center For Bioanalytical Sciences, Department of Biochemistry and Molecular Biology, University of Southern Denmark, DK-5230 Odense M, Denmark.
authors.forename: Jonas
authors.initials: J
authors.lastname: Borch
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Science, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.|bioSYNergy, Center for Synthetic Biology, DK-1871 Frederiksberg C, Denmark.|VILLUM Research Center for Plant Plasticity, DK-1871 Frederiksberg C, Denmark.|Copenhagen Plant Science Center, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.
authors.forename: Camilla
authors.initials: C
authors.lastname: Knudsen
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Science, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.|bioSYNergy, Center for Synthetic Biology, DK-1871 Frederiksberg C, Denmark.|VILLUM Research Center for Plant Plasticity, DK-1871 Frederiksberg C, Denmark.|Copenhagen Plant Science Center, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.
authors.forename: Krutika
authors.initials: K
authors.lastname: Bavishi
authors.affiliation: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597, Singapore.
authors.forename: Federico
authors.initials: F
authors.lastname: Torta
authors.affiliation: Copenhagen Plant Science Center, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.
authors.forename: Helle Juel
authors.initials: HJ
authors.lastname: Martens
authors.affiliation: Copenhagen Plant Science Center, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.
authors.forename: Daniele
authors.initials: D
authors.lastname: Silvestro
authors.affiliation: bioSYNergy, Center for Synthetic Biology, DK-1871 Frederiksberg C, Denmark.|Department of Chemistry, Nano-Science Center, University of Copenhagen, DK-2100 Copenhagen, Denmark.
authors.forename: Nikos S
authors.initials: NS
authors.lastname: Hatzakis
authors.affiliation: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597, Singapore.|Department of Biological Sciences, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597, Singapore.
authors.forename: Markus R
authors.initials: MR
authors.lastname: Wenk
authors.affiliation: School of Biosciences, University of Birmingham, Birmingham B15 2TT, UK.|Department of Business, Energy and Industrial Strategy, Her Majesty's Government, UK.
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Dafforn
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Science, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.|bioSYNergy, Center for Synthetic Biology, DK-1871 Frederiksberg C, Denmark.|VILLUM Research Center for Plant Plasticity, DK-1871 Frederiksberg C, Denmark.
authors.forename: Carl Erik
authors.initials: CE
authors.lastname: Olsen
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Science, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.|bioSYNergy, Center for Synthetic Biology, DK-1871 Frederiksberg C, Denmark.|VILLUM Research Center for Plant Plasticity, DK-1871 Frederiksberg C, Denmark.|Copenhagen Plant Science Center, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.
authors.forename: Mohammed Saddik
authors.initials: MS
authors.lastname: Motawia
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Science, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.|bioSYNergy, Center for Synthetic Biology, DK-1871 Frederiksberg C, Denmark.
authors.forename: Björn
authors.initials: B
authors.lastname: Hamberger
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Science, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.|bioSYNergy, Center for Synthetic Biology, DK-1871 Frederiksberg C, Denmark.|VILLUM Research Center for Plant Plasticity, DK-1871 Frederiksberg C, Denmark.|Copenhagen Plant Science Center, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.|Carlsberg Research Laboratory, DK-1799 Copenhagen V, Denmark.
authors.forename: Birger Lindberg
authors.initials: BL
authors.lastname: Møller
authors.affiliation: Plant Biochemistry Laboratory, Department of Plant and Environmental Science, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.|bioSYNergy, Center for Synthetic Biology, DK-1871 Frederiksberg C, Denmark.|VILLUM Research Center for Plant Plasticity, DK-1871 Frederiksberg C, Denmark.|Copenhagen Plant Science Center, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.
authors.forename: Jean-Etienne
authors.initials: JE
authors.lastname: Bassard
mesh_terms: D055162:Biocatalysis; D053898:Biosynthetic Pathways; D003902:Detergents / Q000737:chemistry; D005964:Glucosyltransferases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D008055:Lipids / Q000737:chemistry / Q000302:isolation & purification; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D008164:Luminescent Proteins / Q000032:analysis / Q000737:chemistry; D009097:Multienzyme Complexes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D009570:Nitriles / Q000378:metabolism*; D061848:Optical Imaging; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D060066:Protein Interaction Maps; D045868:Sorghum / Q000201:enzymology*; D013050:Spectrometry, Fluorescence; D000097573:Red Fluorescent Protein
chemical_list: D003902:Detergents; D005964:Glucosyltransferases; D008055:Lipids; D008081:Liposomes; D008164:Luminescent Proteins; D009097:Multienzyme Complexes; D009570:Nitriles; D010940:Plant Proteins; D000097573:Red Fluorescent Protein; C011220:dhurrin