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Science. 2016 Nov 18;354(6314):890-893.

Characterization of a dynamic metabolon producing the defense compound dhurrin in sorghum.

Laursen T1,2,3,4,5, Borch J2,6, Knudsen C1,2,3,4, Bavishi K1,2,3,4, Torta F7, Martens HJ4, Silvestro D4, Hatzakis NS2,8, Wenk MR7,9, Dafforn TR10,11, Olsen CE1,2,3, Motawia MS1,2,3,4, Hamberger B1,2, Møller BL1,2,3,4,12, Bassard JE1,2,3,4.

Author information

1
Plant Biochemistry Laboratory, Department of Plant and Environmental Science, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.
2
bioSYNergy, Center for Synthetic Biology, DK-1871 Frederiksberg C, Denmark.
3
VILLUM Research Center for Plant Plasticity, DK-1871 Frederiksberg C, Denmark.
4
Copenhagen Plant Science Center, University of Copenhagen, DK-1871 Frederiksberg C, Denmark.
5
Feedstocks Division, Joint BioEnergy Institute, Emeryville, CA 94608, USA.
6
VILLUM Center For Bioanalytical Sciences, Department of Biochemistry and Molecular Biology, University of Southern Denmark, DK-5230 Odense M, Denmark.
7
Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597, Singapore.
8
Department of Chemistry, Nano-Science Center, University of Copenhagen, DK-2100 Copenhagen, Denmark.
9
Department of Biological Sciences, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597, Singapore.
10
School of Biosciences, University of Birmingham, Birmingham B15 2TT, UK.
11
Department of Business, Energy and Industrial Strategy, Her Majesty's Government, UK.
12
Carlsberg Research Laboratory, DK-1799 Copenhagen V, Denmark.

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.

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PMID:
27856908
DOI:
10.1126/science.aag2347
[Indexed for MEDLINE]

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