Engineering Nicotiana benthamiana for chrysoeriol production using synthetic biology approaches

Flavonoids are prevalent plant secondary metabolites with a broad range of biological activities. Their antioxidant, anti-inflammatory, and anti-cancer activities make flavonoids widely useful in a variety of industries, including the pharmaceutical and health food industries. However, many flavonoi...

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Main Authors: Saet Buyl Lee, Sung-eun Lee, Hyo Lee, Ji-Su Kim, Hyoseon Choi, Sichul Lee, Beom-Gi Kim
Format: Article
Language:English
Published: Frontiers Media S.A. 2024-12-01
Series:Frontiers in Plant Science
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Online Access:https://www.frontiersin.org/articles/10.3389/fpls.2024.1458916/full
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author Saet Buyl Lee
Sung-eun Lee
Hyo Lee
Ji-Su Kim
Hyoseon Choi
Sichul Lee
Beom-Gi Kim
author_facet Saet Buyl Lee
Sung-eun Lee
Hyo Lee
Ji-Su Kim
Hyoseon Choi
Sichul Lee
Beom-Gi Kim
author_sort Saet Buyl Lee
collection DOAJ
description Flavonoids are prevalent plant secondary metabolites with a broad range of biological activities. Their antioxidant, anti-inflammatory, and anti-cancer activities make flavonoids widely useful in a variety of industries, including the pharmaceutical and health food industries. However, many flavonoids occur at only low concentrations in plants, and they are difficult to synthesize chemically due to their structural complexity. To address these difficulties, new technologies have been employed to enhance the production of flavonoids in vivo. In this study, we used synthetic biology techniques to produce the methylated flavone chrysoeriol in Nicotiana benthamiana leaves. The chrysoeriol biosynthetic pathway consists of eight catalytic steps. However, using an Agrobacterium-mediated transient expression assay to examine the in planta activities of genes of interest, we shortened this pathway to four steps catalyzed by five enzymes. Co-expression of these five enzymes in N. benthamiana leaves resulted in de novo chrysoeriol production. Chrysoeriol production was unaffected by the Agrobacterium cell density used for agroinfiltration and increased over time, peaking at 10 days after infiltration. Chrysoeriol accumulation in agroinfiltrated N. benthamiana leaves was associated with increased antioxidant activity, a typical property of flavones. Taken together, our results demonstrate that synthetic biology represents a practical method for engineering plants to produce substantial amounts of flavonoids and flavonoid derivatives without the need for exogenous substrates.
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spelling doaj-art-e860a8f5c3f04d109db6a6b8e03ab2072025-08-20T02:36:54ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2024-12-011510.3389/fpls.2024.14589161458916Engineering Nicotiana benthamiana for chrysoeriol production using synthetic biology approachesSaet Buyl LeeSung-eun LeeHyo LeeJi-Su KimHyoseon ChoiSichul LeeBeom-Gi KimFlavonoids are prevalent plant secondary metabolites with a broad range of biological activities. Their antioxidant, anti-inflammatory, and anti-cancer activities make flavonoids widely useful in a variety of industries, including the pharmaceutical and health food industries. However, many flavonoids occur at only low concentrations in plants, and they are difficult to synthesize chemically due to their structural complexity. To address these difficulties, new technologies have been employed to enhance the production of flavonoids in vivo. In this study, we used synthetic biology techniques to produce the methylated flavone chrysoeriol in Nicotiana benthamiana leaves. The chrysoeriol biosynthetic pathway consists of eight catalytic steps. However, using an Agrobacterium-mediated transient expression assay to examine the in planta activities of genes of interest, we shortened this pathway to four steps catalyzed by five enzymes. Co-expression of these five enzymes in N. benthamiana leaves resulted in de novo chrysoeriol production. Chrysoeriol production was unaffected by the Agrobacterium cell density used for agroinfiltration and increased over time, peaking at 10 days after infiltration. Chrysoeriol accumulation in agroinfiltrated N. benthamiana leaves was associated with increased antioxidant activity, a typical property of flavones. Taken together, our results demonstrate that synthetic biology represents a practical method for engineering plants to produce substantial amounts of flavonoids and flavonoid derivatives without the need for exogenous substrates.https://www.frontiersin.org/articles/10.3389/fpls.2024.1458916/fullchrysoeriolco-expressionflavonoidgene combinationNicotiana benthamianareconstruction
spellingShingle Saet Buyl Lee
Sung-eun Lee
Hyo Lee
Ji-Su Kim
Hyoseon Choi
Sichul Lee
Beom-Gi Kim
Engineering Nicotiana benthamiana for chrysoeriol production using synthetic biology approaches
Frontiers in Plant Science
chrysoeriol
co-expression
flavonoid
gene combination
Nicotiana benthamiana
reconstruction
title Engineering Nicotiana benthamiana for chrysoeriol production using synthetic biology approaches
title_full Engineering Nicotiana benthamiana for chrysoeriol production using synthetic biology approaches
title_fullStr Engineering Nicotiana benthamiana for chrysoeriol production using synthetic biology approaches
title_full_unstemmed Engineering Nicotiana benthamiana for chrysoeriol production using synthetic biology approaches
title_short Engineering Nicotiana benthamiana for chrysoeriol production using synthetic biology approaches
title_sort engineering nicotiana benthamiana for chrysoeriol production using synthetic biology approaches
topic chrysoeriol
co-expression
flavonoid
gene combination
Nicotiana benthamiana
reconstruction
url https://www.frontiersin.org/articles/10.3389/fpls.2024.1458916/full
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AT jisukim engineeringnicotianabenthamianaforchrysoeriolproductionusingsyntheticbiologyapproaches
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