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dc.contributor.authorDavis, Katharine
dc.contributor.authorGkotsi, Danai S.
dc.contributor.authorSmith, Duncan R. M.
dc.contributor.authorGoss, Rebecca J. M.
dc.contributor.authorCaputi, Lorenzo
dc.contributor.authorO’Connor, Sarah E.
dc.date.accessioned2020-12-04T05:06:02Z
dc.date.available2020-12-04T05:06:02Z
dc.date.issued2020-11-20
dc.date.submitted2020-07-09
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/314700
dc.description.abstractPlant secondary metabolites have applications for the food, biofuel, and pharmaceutical industries. Recent advances in pathway elucidation and host expression systems now allow metabolic engineering of plant metabolic pathways to produce “new-to-nature” derivatives with novel biological activities, thereby amplifying the range of industrial uses for plant metabolites. Here we use a transient expression system in the model plant Nicotiana benthamiana to reconstitute the two-step plant-derived biosynthetic pathway for auxin (indole acetic acid) to achieve accumulation up to 500 ng/g fresh mass (FM). By expressing these plant-derived enzymes in combination with either bacterial halogenases and alternative substrates, we can produce both natural and new-to-nature halogenated auxin derivatives up to 990 ng/g FM. Proteins from the auxin synthesis pathway, tryptophan aminotransferases (TARs) and flavin-dependent monooxygenases (YUCs), could be transiently expressed in combination with four separate bacterial halogenases to generate halogenated auxin derivatives. Brominated auxin derivatives could also be observed after infiltration of the transfected N. benthamiana with potassium bromide and the halogenases. Finally, the production of additional auxin derivatives could also be achieved by co-infiltration of TAR and YUC genes with various tryptophan analogs. Given the emerging importance of transient expression in N. benthamiana for industrial scale protein and product expression, this work provides insight into the capacity of N. benthamiana to interface bacterial genes and synthetic substrates to produce novel halogenated metabolites.
dc.languageen
dc.publisherFrontiers Media S.A.
dc.rightsAttribution 4.0 International (CC BY 4.0)en
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/en
dc.subjectPlant Science
dc.subjectindole-acetic acid
dc.subjecthalogenase
dc.subjectcombinatorial biosynthesis
dc.subjectauxin
dc.subjectunnatural natural product
dc.subjectnew to nature products
dc.titleNicotiana benthamiana as a Transient Expression Host to Produce Auxin Analogs
dc.typeArticle
dc.date.updated2020-12-04T05:06:02Z
prism.publicationNameFrontiers in Plant Science
prism.volume11
dc.identifier.doi10.17863/CAM.61804
dcterms.dateAccepted2020-10-26
rioxxterms.versionofrecord10.3389/fpls.2020.581675
rioxxterms.versionVoR
rioxxterms.licenseref.urihttp://creativecommons.org/licenses/by/4.0/
dc.identifier.eissn1664-462X


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Attribution 4.0 International (CC BY 4.0)
Except where otherwise noted, this item's licence is described as Attribution 4.0 International (CC BY 4.0)