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

• Published in: Science (American Association for the Advancement of Science) 2016-11, Vol.354 (6314), p.890-893
• Main Authors: Laursen, Tomas, Borch, Jonas, Knudsen, Camilla, Bavishi, Krutika, Torta, Federico, Martens, Helle Juel, Silvestro, Daniele, Hatzakis, Nikos S., Wenk, Markus R., Dafforn, Timothy R., Olsen, Carl Erik, Motawia, Mohammed Saddik, Hamberger, Björn, Møller, Birger Lindberg, Bassard, Jean-Etienne
• Format: Article
• Language: English
• Subjects: Assembly; Biocatalysis; Biosynthetic Pathways; Channeling; Detergents - chemistry; Dynamics; Enzymes; Fluorescence; Genotype & phenotype; Glucosyltransferases - chemistry; Glucosyltransferases - isolation & purification; Glucosyltransferases - metabolism; Life Sciences; Lipids - chemistry; Lipids - isolation & purification; Liposomes - chemistry; Liposomes - metabolism; Luminescent Proteins - analysis; Luminescent Proteins - chemistry; Metabolic syndrome; Metabolites; Multienzyme Complexes - chemistry; Multienzyme Complexes - isolation & purification; Multienzyme Complexes - metabolism; Nitriles - metabolism; Optical Imaging; Plant Proteins - chemistry; Plant Proteins - isolation & purification; Plant Proteins - metabolism; Plants (organisms); Protein Interaction Maps; Red Fluorescent Protein; Sorghum; Sorghum - enzymology; Spectrometry, Fluorescence; Vegetal Biology
• ISSN: 0036-8075; 1095-9203; 1095-9203
• DOI: 10.1126/science.aag2347

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.