In planta complementation of the betalain biosynthetic pathway with a bacterial dioxygenase.

Betalains are pigments naturally produced by plants belonging to the order Caryophyllales. However, 4,5-L-DOPA-extradiol-dioxygenase (DODA), the main enzyme in their biosynthetic pathway, has also been characterized in other kingdoms including fungi or bacteria. These enzymes show few similarities w...

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Main Authors: María Alejandra Guerrero-Rubio, Hester Sheehan, Samuel F Brockington
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2025-01-01
Series:PLoS ONE
Online Access:https://doi.org/10.1371/journal.pone.0325603
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author María Alejandra Guerrero-Rubio
Hester Sheehan
Samuel F Brockington
author_facet María Alejandra Guerrero-Rubio
Hester Sheehan
Samuel F Brockington
author_sort María Alejandra Guerrero-Rubio
collection DOAJ
description Betalains are pigments naturally produced by plants belonging to the order Caryophyllales. However, 4,5-L-DOPA-extradiol-dioxygenase (DODA), the main enzyme in their biosynthetic pathway, has also been characterized in other kingdoms including fungi or bacteria. These enzymes show few similarities with the plant enzymes at structural or kinetic levels but achieve the same activity, raising questions on their possible exchange with plant DODA enzymes and subsequent complementation in the biosynthetic pathway of betalains. In this work, we show that the DODA enzyme from the bacterium Gluconacetobacter diazotrophicus is able to provide the activity of L-DOPA dioxygenase and produce betalamic acid and betalains, in planta. Betalains were detected after transient expression of the genes required for their synthesis in the plant model Nicotiana benthamiana. HPLC-ESI-MS analysis showed that the expression of the bacterial enzyme gave rise to multiple betalains in a similar manner to plant DODAs, with vulgaxanthin I being the main pigment obtained in infiltrated leaves. Our results show that fully functional betalamic-forming enzymes from outside of the plant kingdom are able to complement the plant-based betalain biosynthetic pathway. This supports the proposition of convergent evolution of enzymes with betalain-forming activity, not only within Caryophyllales, but also in other kingdoms.
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spelling doaj-art-853e02867d004fec96308539e5d92e6a2025-08-20T03:24:44ZengPublic Library of Science (PLoS)PLoS ONE1932-62032025-01-01206e032560310.1371/journal.pone.0325603In planta complementation of the betalain biosynthetic pathway with a bacterial dioxygenase.María Alejandra Guerrero-RubioHester SheehanSamuel F BrockingtonBetalains are pigments naturally produced by plants belonging to the order Caryophyllales. However, 4,5-L-DOPA-extradiol-dioxygenase (DODA), the main enzyme in their biosynthetic pathway, has also been characterized in other kingdoms including fungi or bacteria. These enzymes show few similarities with the plant enzymes at structural or kinetic levels but achieve the same activity, raising questions on their possible exchange with plant DODA enzymes and subsequent complementation in the biosynthetic pathway of betalains. In this work, we show that the DODA enzyme from the bacterium Gluconacetobacter diazotrophicus is able to provide the activity of L-DOPA dioxygenase and produce betalamic acid and betalains, in planta. Betalains were detected after transient expression of the genes required for their synthesis in the plant model Nicotiana benthamiana. HPLC-ESI-MS analysis showed that the expression of the bacterial enzyme gave rise to multiple betalains in a similar manner to plant DODAs, with vulgaxanthin I being the main pigment obtained in infiltrated leaves. Our results show that fully functional betalamic-forming enzymes from outside of the plant kingdom are able to complement the plant-based betalain biosynthetic pathway. This supports the proposition of convergent evolution of enzymes with betalain-forming activity, not only within Caryophyllales, but also in other kingdoms.https://doi.org/10.1371/journal.pone.0325603
spellingShingle María Alejandra Guerrero-Rubio
Hester Sheehan
Samuel F Brockington
In planta complementation of the betalain biosynthetic pathway with a bacterial dioxygenase.
PLoS ONE
title In planta complementation of the betalain biosynthetic pathway with a bacterial dioxygenase.
title_full In planta complementation of the betalain biosynthetic pathway with a bacterial dioxygenase.
title_fullStr In planta complementation of the betalain biosynthetic pathway with a bacterial dioxygenase.
title_full_unstemmed In planta complementation of the betalain biosynthetic pathway with a bacterial dioxygenase.
title_short In planta complementation of the betalain biosynthetic pathway with a bacterial dioxygenase.
title_sort in planta complementation of the betalain biosynthetic pathway with a bacterial dioxygenase
url https://doi.org/10.1371/journal.pone.0325603
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AT samuelfbrockington inplantacomplementationofthebetalainbiosyntheticpathwaywithabacterialdioxygenase