Reconstitution of monoterpene indole alkaloid biosynthesis in genome engineered Nicotiana benthamiana.

Autor: Dudley QM; Engineering Biology, Earlham Institute, Norwich Research Park, Norwich, Norfolk, NR4 7UZ, UK., Jo S; Engineering Biology, Earlham Institute, Norwich Research Park, Norwich, Norfolk, NR4 7UZ, UK., Guerrero DAS; Department of Natural Product Biosynthesis, Max Planck Institute for Chemical Ecology, Jena, 07745, Germany., Chhetry M; John Innes Centre, Norwich Research Park, Norwich, NR4 7TJ, UK., Smedley MA; John Innes Centre, Norwich Research Park, Norwich, NR4 7TJ, UK., Harwood WA; John Innes Centre, Norwich Research Park, Norwich, NR4 7TJ, UK., Sherden NH; John Innes Centre, Norwich Research Park, Norwich, NR4 7TJ, UK.; Octagon Therapeutics Ltd, 700 Main Street, North Cambridge, MA, 02139, USA., O'Connor SE; Department of Natural Product Biosynthesis, Max Planck Institute for Chemical Ecology, Jena, 07745, Germany., Caputi L; Department of Natural Product Biosynthesis, Max Planck Institute for Chemical Ecology, Jena, 07745, Germany. lcaputi@ice.mpg.de., Patron NJ; Engineering Biology, Earlham Institute, Norwich Research Park, Norwich, Norfolk, NR4 7UZ, UK. nicola.patron@earlham.ac.uk.
Jazyk: angličtina
Zdroj: Communications biology [Commun Biol] 2022 Sep 10; Vol. 5 (1), pp. 949. Date of Electronic Publication: 2022 Sep 10.
DOI: 10.1038/s42003-022-03904-w
Abstrakt: Monoterpene indole alkaloids (MIAs) are a diverse class of plant natural products that include a number of medicinally important compounds. We set out to reconstitute the pathway for strictosidine, a key intermediate of all MIAs, from central metabolism in Nicotiana benthamiana. A disadvantage of this host is that its rich background metabolism results in the derivatization of some heterologously produced molecules. Here we use transcriptomic analysis to identify glycosyltransferases that are upregulated in response to biosynthetic intermediates and produce plant lines with targeted mutations in the genes encoding them. Expression of the early MIA pathway in these lines produces a more favorable product profile. Strictosidine biosynthesis was successfully reconstituted, with the best yields obtained by the co-expression of 14 enzymes, of which a major latex protein-like enzyme (MLPL) from Nepeta (catmint) is critical for improving flux through the iridoid pathway. The removal of endogenous glycosyltransferases does not impact the yields of strictosidine, highlighting that the metabolic flux of the pathway enzymes to a stable biosynthetic intermediate minimizes the need to engineer the endogenous metabolism of the host. The production of strictosidine in planta expands the range of MIA products amenable to biological synthesis.
(© 2022. The Author(s).)
Databáze: MEDLINE
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