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Exploiting the Substrate Promiscuity of Hydroxycinnamoyl-CoA:Shikimate Hydroxycinnamoyl Transferase to Reduce Lignin

Lignin poses a major challenge in the processing of plant biomass for agro-industrial applications. For bioengineering purposes, there is a pressing interest in identifying and characterizing the enzymes responsible for the biosynthesis of lignin. Hydroxycinnamoyl-CoA:shikimate hydroxycinnamoyl tran...

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Autores principales: Eudes, Aymerick, Pereira, Jose H., Yogiswara, Sasha, Wang, George, Teixeira Benites, Veronica, Baidoo, Edward E.K., Lee, Taek Soon, Adams, Paul D., Keasling, Jay D., Loqué, Dominique
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4790474/
https://www.ncbi.nlm.nih.gov/pubmed/26858288
http://dx.doi.org/10.1093/pcp/pcw016
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author Eudes, Aymerick
Pereira, Jose H.
Yogiswara, Sasha
Wang, George
Teixeira Benites, Veronica
Baidoo, Edward E.K.
Lee, Taek Soon
Adams, Paul D.
Keasling, Jay D.
Loqué, Dominique
author_facet Eudes, Aymerick
Pereira, Jose H.
Yogiswara, Sasha
Wang, George
Teixeira Benites, Veronica
Baidoo, Edward E.K.
Lee, Taek Soon
Adams, Paul D.
Keasling, Jay D.
Loqué, Dominique
author_sort Eudes, Aymerick
collection PubMed
description Lignin poses a major challenge in the processing of plant biomass for agro-industrial applications. For bioengineering purposes, there is a pressing interest in identifying and characterizing the enzymes responsible for the biosynthesis of lignin. Hydroxycinnamoyl-CoA:shikimate hydroxycinnamoyl transferase (HCT; EC 2.3.1.133) is a key metabolic entry point for the synthesis of the most important lignin monomers: coniferyl and sinapyl alcohols. In this study, we investigated the substrate promiscuity of HCT from a bryophyte (Physcomitrella) and from five representatives of vascular plants (Arabidopsis, poplar, switchgrass, pine and Selaginella) using a yeast expression system. We demonstrate for these HCTs a conserved capacity to acylate with p-coumaroyl-CoA several phenolic compounds in addition to the canonical acceptor shikimate normally used during lignin biosynthesis. Using either recombinant HCT from switchgrass (PvHCT2a) or an Arabidopsis stem protein extract, we show evidence of the inhibitory effect of these phenolics on the synthesis of p-coumaroyl shikimate in vitro, which presumably occurs via a mechanism of competitive inhibition. A structural study of PvHCT2a confirmed the binding of a non-canonical acceptor in a similar manner to shikimate in the active site of the enzyme. Finally, we exploited in Arabidopsis the substrate flexibility of HCT to reduce lignin content and improve biomass saccharification by engineering transgenic lines that overproduce one of the HCT non-canonical acceptors. Our results demonstrate conservation of HCT substrate promiscuity and provide support for a new strategy for lignin reduction in the effort to improve the quality of plant biomass for forage and cellulosic biofuels.
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spelling pubmed-47904742016-03-16 Exploiting the Substrate Promiscuity of Hydroxycinnamoyl-CoA:Shikimate Hydroxycinnamoyl Transferase to Reduce Lignin Eudes, Aymerick Pereira, Jose H. Yogiswara, Sasha Wang, George Teixeira Benites, Veronica Baidoo, Edward E.K. Lee, Taek Soon Adams, Paul D. Keasling, Jay D. Loqué, Dominique Plant Cell Physiol Regular Papers Lignin poses a major challenge in the processing of plant biomass for agro-industrial applications. For bioengineering purposes, there is a pressing interest in identifying and characterizing the enzymes responsible for the biosynthesis of lignin. Hydroxycinnamoyl-CoA:shikimate hydroxycinnamoyl transferase (HCT; EC 2.3.1.133) is a key metabolic entry point for the synthesis of the most important lignin monomers: coniferyl and sinapyl alcohols. In this study, we investigated the substrate promiscuity of HCT from a bryophyte (Physcomitrella) and from five representatives of vascular plants (Arabidopsis, poplar, switchgrass, pine and Selaginella) using a yeast expression system. We demonstrate for these HCTs a conserved capacity to acylate with p-coumaroyl-CoA several phenolic compounds in addition to the canonical acceptor shikimate normally used during lignin biosynthesis. Using either recombinant HCT from switchgrass (PvHCT2a) or an Arabidopsis stem protein extract, we show evidence of the inhibitory effect of these phenolics on the synthesis of p-coumaroyl shikimate in vitro, which presumably occurs via a mechanism of competitive inhibition. A structural study of PvHCT2a confirmed the binding of a non-canonical acceptor in a similar manner to shikimate in the active site of the enzyme. Finally, we exploited in Arabidopsis the substrate flexibility of HCT to reduce lignin content and improve biomass saccharification by engineering transgenic lines that overproduce one of the HCT non-canonical acceptors. Our results demonstrate conservation of HCT substrate promiscuity and provide support for a new strategy for lignin reduction in the effort to improve the quality of plant biomass for forage and cellulosic biofuels. Oxford University Press 2016-03 2016-02-08 /pmc/articles/PMC4790474/ /pubmed/26858288 http://dx.doi.org/10.1093/pcp/pcw016 Text en © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Regular Papers
Eudes, Aymerick
Pereira, Jose H.
Yogiswara, Sasha
Wang, George
Teixeira Benites, Veronica
Baidoo, Edward E.K.
Lee, Taek Soon
Adams, Paul D.
Keasling, Jay D.
Loqué, Dominique
Exploiting the Substrate Promiscuity of Hydroxycinnamoyl-CoA:Shikimate Hydroxycinnamoyl Transferase to Reduce Lignin
title Exploiting the Substrate Promiscuity of Hydroxycinnamoyl-CoA:Shikimate Hydroxycinnamoyl Transferase to Reduce Lignin
title_full Exploiting the Substrate Promiscuity of Hydroxycinnamoyl-CoA:Shikimate Hydroxycinnamoyl Transferase to Reduce Lignin
title_fullStr Exploiting the Substrate Promiscuity of Hydroxycinnamoyl-CoA:Shikimate Hydroxycinnamoyl Transferase to Reduce Lignin
title_full_unstemmed Exploiting the Substrate Promiscuity of Hydroxycinnamoyl-CoA:Shikimate Hydroxycinnamoyl Transferase to Reduce Lignin
title_short Exploiting the Substrate Promiscuity of Hydroxycinnamoyl-CoA:Shikimate Hydroxycinnamoyl Transferase to Reduce Lignin
title_sort exploiting the substrate promiscuity of hydroxycinnamoyl-coa:shikimate hydroxycinnamoyl transferase to reduce lignin
topic Regular Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4790474/
https://www.ncbi.nlm.nih.gov/pubmed/26858288
http://dx.doi.org/10.1093/pcp/pcw016
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