skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

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

Abstract

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 biomassmore » 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.« less

Authors:
 [1];  [2];  [3];  [1];  [4];  [1];  [1];  [2];  [3];  [1]
  1. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering Division
  2. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Molecular Biophysics and Integrated Bioimaging Division
  3. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering Division; Univ. of California, Berkeley, CA (United States). Dept. of Bioengineering & Dept. of Chemical & Biomolecular Engineering
  4. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering Division; San Francisco State Univ., CA (United States)
Publication Date:
Research Org.:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER); National Institutes of Health (NIH); USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1379120
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Plant and Cell Physiology
Additional Journal Information:
Journal Volume: 57; Journal Issue: 3; Journal ID: ISSN 0032-0781
Publisher:
Japanese Society of Plant Physiologists
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 09 BIOMASS FUELS; Arabidopsis; Bioenergy; Cell wall; HCT; Lignin; Saccharification

Citation Formats

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., and Loqué, Dominique. Exploiting the Substrate Promiscuity of Hydroxycinnamoyl-CoA:Shikimate Hydroxycinnamoyl Transferase to Reduce Lignin. United States: N. p., 2016. Web. doi:10.1093/pcp/pcw016.
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. United States. https://doi.org/10.1093/pcp/pcw016
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., and Loqué, Dominique. 2016. "Exploiting the Substrate Promiscuity of Hydroxycinnamoyl-CoA:Shikimate Hydroxycinnamoyl Transferase to Reduce Lignin". United States. https://doi.org/10.1093/pcp/pcw016. https://www.osti.gov/servlets/purl/1379120.
@article{osti_1379120,
title = {Exploiting the Substrate Promiscuity of Hydroxycinnamoyl-CoA:Shikimate Hydroxycinnamoyl Transferase to Reduce Lignin},
author = {Eudes, Aymerick and Pereira, Jose H. and Yogiswara, Sasha and Wang, George and Teixeira Benites, Veronica and Baidoo, Edward E. K. and Lee, Taek Soon and Adams, Paul D. and Keasling, Jay D. and Loqué, Dominique},
abstractNote = {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.},
doi = {10.1093/pcp/pcw016},
url = {https://www.osti.gov/biblio/1379120}, journal = {Plant and Cell Physiology},
issn = {0032-0781},
number = 3,
volume = 57,
place = {United States},
year = {Mon Feb 08 00:00:00 EST 2016},
month = {Mon Feb 08 00:00:00 EST 2016}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Citation Metrics:
Cited by: 56 works
Citation information provided by
Web of Science

Save / Share:

Works referenced in this record:

Biosynthesis of the immunosuppressants FK506, FK520, and rapamycin involves a previously undescribed family of enzymes acting on chorismate
journal, March 2011


PHENIX: a comprehensive Python-based system for macromolecular structure solution
journal, January 2010


Towards automated crystallographic structure refinement with phenix.refine
journal, March 2012


Lignin Biosynthesis
journal, June 2003


Lignin modification improves fermentable sugar yields for biofuel production
journal, June 2007


Oat phenolics: avenanthramides, novel substituted N-cinnamoylanthranilate alkaloids from oat groats and hulls
journal, January 1989


Acyltransferases in plants: a good time to be BAHD
journal, June 2006


MolProbity: all-atom contacts and structure validation for proteins and nucleic acids
journal, May 2007


Structural Characterization of Wheat Straw Lignin as Revealed by Analytical Pyrolysis, 2D-NMR, and Reductive Cleavage Methods
journal, December 2011


Small Glycosylated Lignin Oligomers Are Stored in Arabidopsis Leaf Vacuoles
journal, February 2015


Early lignin pathway enzymes and routes to chlorogenic acid in switchgrass (Panicum virgatum L.)
journal, November 2013


Production of tranilast [N-(3′,4′-dimethoxycinnamoyl)-anthranilic acid] and its analogs in yeast Saccharomyces cerevisiae
journal, October 2010


Production of hydroxycinnamoyl anthranilates from glucose in Escherichia coli
journal, January 2013


Expression of a bacterial 3-dehydroshikimate dehydratase reduces lignin content and improves biomass saccharification efficiency
journal, January 2015


Metabolite profiling of the moss Physcomitrella patens reveals evolutionary conservation of osmoprotective substances
journal, October 2011


Silencing of Hydroxycinnamoyl-Coenzyme A Shikimate/Quinate Hydroxycinnamoyltransferase Affects Phenylpropanoid Biosynthesis
journal, May 2004


Purification, Cloning, and Properties of an Acyltransferase Controlling Shikimate and Quinate Ester Intermediates in Phenylpropanoid Metabolism
journal, October 2002


Sparse matrix sampling: a screening method for crystallization of proteins
journal, August 1991


Characterization of hydroxycinnamoyl-coenzyme a Shikimate hydroxycinnamoyltransferase from Populus euramericana
journal, October 2011


A Structural Basis for the Biosynthesis of the Major Chlorogenic Acids Found in Coffee
journal, July 2012


Substrate promiscuity of a rosmarinic acid synthase from lavender (Lavandula angustifolia L.)
journal, March 2011


Lignin valorization through integrated biological funneling and chemical catalysis
journal, August 2014


Phaser crystallographic software
journal, July 2007


Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugar
journal, March 1959


Role of HXXXD-motif/BAHD acyltransferases in the biosynthesis of extracellular lipids
journal, December 2014


NMRPipe: A multidimensional spectral processing system based on UNIX pipes
journal, November 1995


Salicylic acid beyond defence: its role in plant growth and development
journal, February 2011


Quantitative Determination of Arbutin and Hydroquinone in Different Plant Materials by HPLC
journal, November 2012


A Genomics Approach to Deciphering Lignin Biosynthesis in Switchgrass
journal, November 2013


MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0
journal, October 2013


The evolution of phenylpropanoid metabolism in the green lineage
journal, December 2012


Breeding with rare defective alleles (BRDA): a natural Populus nigra HCT mutant with modified lignin as a case study
journal, February 2013


Caffeoyl Shikimate Esterase (CSE) Is an Enzyme in the Lignin Biosynthetic Pathway in Arabidopsis
journal, August 2013


Examining the feasibility of bulk commodity production in Escherichia coli
journal, December 2011


Exploring lignification in conifers by silencing hydroxycinnamoyl-CoA:shikimate hydroxycinnamoyltransferase in Pinus radiata
journal, July 2007


Convergent Evolution of Syringyl Lignin Biosynthesis via Distinct Pathways in the Lycophyte Selaginella and Flowering Plants
journal, April 2010


Lignin plays a negative role in the biochemical process for producing lignocellulosic biofuels
journal, June 2014


Salicylic acid 3-hydroxylase regulates Arabidopsis leaf longevity by mediating salicylic acid catabolism
journal, August 2013


Cavin1 intrinsically disordered domains are essential for fuzzy electrostatic interactions and caveola formation
journal, February 2021


Multi-functionality of a tryptophan residue conserved in substrate-binding groove of GH19 chitinases
journal, January 2021


MolProbity: all-atom contacts and structure validation for proteins and nucleic acids
text, January 2007


PHENIX: a comprehensive Python-based system for macromolecular structure solution.
text, January 2010


Production of tranilast [N-(3′,4′-dimethoxycinnamoyl)-anthranilic acid] and its analogs in yeast Saccharomyces cerevisiae
journal, October 2010


Metabolite profiling of the moss Physcomitrella patens reveals evolutionary conservation of osmoprotective substances
journal, October 2011


Role of HXXXD-motif/BAHD acyltransferases in the biosynthesis of extracellular lipids
journal, December 2014


Substrate promiscuity of a rosmarinic acid synthase from lavender (Lavandula angustifolia L.)
journal, March 2011


Examining the feasibility of bulk commodity production in Escherichia coli
journal, December 2011


Early lignin pathway enzymes and routes to chlorogenic acid in switchgrass (Panicum virgatum L.)
journal, November 2013


Lignin plays a negative role in the biochemical process for producing lignocellulosic biofuels
journal, June 2014


Acyltransferases in plants: a good time to be BAHD
journal, June 2006


Loosening lignin's grip on biofuel production
journal, July 2007


Changes in secondary metabolism and deposition of an unusual lignin in the ref8 mutant of Arabidopsis
journal, April 2002


Exploring lignification in conifers by silencing hydroxycinnamoyl-CoA:shikimate hydroxycinnamoyltransferase in Pinus radiata
journal, July 2007


Lignin valorization through integrated biological funneling and chemical catalysis
journal, August 2014


Increase in 4-Coumaryl Alcohol Units during Lignification in Alfalfa ( Medicago sativa ) Alters the Extractability and Molecular Weight of Lignin
journal, October 2010


Salicylic acid beyond defence: its role in plant growth and development
journal, February 2011


MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0
journal, October 2013


Silencing of Hydroxycinnamoyl-Coenzyme A Shikimate/Quinate Hydroxycinnamoyltransferase Affects Phenylpropanoid Biosynthesis
journal, May 2004


Convergent Evolution of Syringyl Lignin Biosynthesis via Distinct Pathways in the Lycophyte Selaginella and Flowering Plants
journal, April 2010


Breaking Down the Complex Regulatory Web Underlying Lignin Biosynthesis
journal, November 2013


Small Glycosylated Lignin Oligomers Are Stored in Arabidopsis Leaf Vacuoles
journal, February 2015


Coot model-building tools for molecular graphics
journal, November 2004


Next-generation sequencing for next-generation breeding, and more
journal, April 2013


Expression of a bacterial 3-dehydroshikimate dehydratase reduces lignin content and improves biomass saccharification efficiency
journal, January 2015


Caffeoyl Shikimate Esterase (CSE) Is an Enzyme in the Lignin Biosynthetic Pathway in Arabidopsis
journal, August 2013


Lignin Biosynthesis
journal, June 2003


Comparative genome analysis of lignin biosynthesis gene families across the plant kingdom
journal, January 2009


Production of hydroxycinnamoyl anthranilates from glucose in Escherichia coli
journal, January 2013


Quantitative Determination of Arbutin and Hydroquinone in Different Plant Materials by HPLC
journal, November 2012


PHENIX: a comprehensive Python-based system for macromolecular structure solution.
text, January 2010


Works referencing / citing this record:

Evolution of coumaroyl conjugate 3‐hydroxylases in land plants: lignin biosynthesis and defense
journal, June 2019


Exploiting members of the BAHD acyltransferase family to synthesize multiple hydroxycinnamate and benzoate conjugates in yeast
journal, November 2016


Gene stacking of multiple traits for high yield of fermentable sugars in plant biomass
journal, January 2018


A screening method to identify efficient sgRNAs in Arabidopsis, used in conjunction with cell-specific lignin reduction
journal, May 2019


Molecular Identification and Characterization of Hydroxycinnamoyl Transferase in Tea Plants (Camellia sinensis L.)
journal, December 2018


Rhizobacteria Mediate the Phytotoxicity of a Range of Biorefinery‐Relevant Compounds
journal, July 2019


Exploiting members of the BAHD acyltransferase family to synthesize multiple hydroxycinnamate and benzoate conjugates in yeast
journal, November 2016


Gene stacking of multiple traits for high yield of fermentable sugars in plant biomass
journal, January 2018


Xylan in the Middle: Understanding Xylan Biosynthesis and Its Metabolic Dependencies Toward Improving Wood Fiber for Industrial Processing
journal, February 2019


Molecular Identification and Characterization of Hydroxycinnamoyl Transferase in Tea Plants (Camellia sinensis L.)
journal, December 2018