Heterologous expression of Arabidopsis laccase2, laccase4 and peroxidase52 driven under developing xylem specific promoter DX15 improves saccharification in populus
Abstract
Secondary cell wall holds considerable potential as it has gained immense momentum to replace the lignocellulosic feedstock into fuels. Lignin one of the components of secondary cell wall tightly holds the polysaccharides thereby enhancing the recalcitrance and complexity in the biomass. Laccases (LAC) and peroxidases (PRX) are the major phenyl-oxidases playing key functions during the polymerization of monolignols into lignin. Yet, the functions of laccase and peroxidases gene families remained largely unknown. Hence, the objective of this conducted study is to understand the role of specific LAC and PRX in Populus wood formation and to further investigate how the altered Lac and Prx expression affects biomass recalcitrance and plant growth. This study of heterologous expression of Arabidopsis Lac and Prx genes was conducted in poplar to avoid any otherwise occurring co-suppression mechanism during the homologous overexpression of highly expressed native genes. In the pursuit of optimizing lignocellulosic biomass for biofuel production, the present study focuses on harnessing the enzymatic potential of Arabidopsis thaliana Laccase2, Laccase4, and Peroxidase52 through heterologous expression. We overexpressed selected Arabidopsis laccase2 (AtLac2), laccase4 (AtLac4), and peroxidase52 (AtPrx52) genes, based on their high transcript expression respective to the differentiating xylem tissues in the stem, in hybrid poplarmore »
- Authors:
-
- Michigan Technological University, Houghton, MI (United States)
- University of Georgia, Athens, GA (United States)
- Universiti Kebangsaan Malaysia (UKM) Bangi (Malaysia)
- National Renewable Energy Laboratory (NREL), Golden, CO (United States)
- Dr. Yashwant Singh Parmar University of Horticulture and Forestry (YSP UHF), Nauni (India)
- College of Agriculture, Science, and Technology (CAST), Dover, DE (United States)
- Publication Date:
- Research Org.:
- National Renewable Energy Laboratory (NREL), Golden, CO (United States); Michigan Technological Univ., Houghton, MI (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER); Ministry of Science and Technology (MoST); National Science Foundation (NSF)
- OSTI Identifier:
- 2283933
- Report Number(s):
- NREL/JA-2800-88719
Journal ID: ISSN 2731-3654; MainId:89498;UUID:d2700bfe-e485-4e2a-b813-d4c9b2289db4;MainAdminId:71731
- Grant/Contract Number:
- AC36-08GO28308; 1230803; R31-2009-000-20025-0
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Biotechnology for Biofuels and Bioproducts
- Additional Journal Information:
- Journal Volume: 17; Journal Issue: 1; Journal ID: ISSN 2731-3654
- Publisher:
- BioMed Central
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 09 BIOMASS FUELS; arabidopsis; bioethanol; developing xylem; heterologous expression; laccases; peroxidases; poplars; saccharification; secondary cell walls
Citation Formats
Ahlawat, Yogesh K., Biswal, Ajaya K., Harun, Sarahani, Harman-Ware, Anne E., Doeppke, Crissa, Sharma, Nisha, Joshi, Chandrashekhar P., and Hankoua, Bertrand B. Heterologous expression of Arabidopsis laccase2, laccase4 and peroxidase52 driven under developing xylem specific promoter DX15 improves saccharification in populus. United States: N. p., 2024.
Web. doi:10.1186/s13068-023-02452-7.
Ahlawat, Yogesh K., Biswal, Ajaya K., Harun, Sarahani, Harman-Ware, Anne E., Doeppke, Crissa, Sharma, Nisha, Joshi, Chandrashekhar P., & Hankoua, Bertrand B. Heterologous expression of Arabidopsis laccase2, laccase4 and peroxidase52 driven under developing xylem specific promoter DX15 improves saccharification in populus. United States. https://doi.org/10.1186/s13068-023-02452-7
Ahlawat, Yogesh K., Biswal, Ajaya K., Harun, Sarahani, Harman-Ware, Anne E., Doeppke, Crissa, Sharma, Nisha, Joshi, Chandrashekhar P., and Hankoua, Bertrand B. Sat .
"Heterologous expression of Arabidopsis laccase2, laccase4 and peroxidase52 driven under developing xylem specific promoter DX15 improves saccharification in populus". United States. https://doi.org/10.1186/s13068-023-02452-7. https://www.osti.gov/servlets/purl/2283933.
@article{osti_2283933,
title = {Heterologous expression of Arabidopsis laccase2, laccase4 and peroxidase52 driven under developing xylem specific promoter DX15 improves saccharification in populus},
author = {Ahlawat, Yogesh K. and Biswal, Ajaya K. and Harun, Sarahani and Harman-Ware, Anne E. and Doeppke, Crissa and Sharma, Nisha and Joshi, Chandrashekhar P. and Hankoua, Bertrand B.},
abstractNote = {Secondary cell wall holds considerable potential as it has gained immense momentum to replace the lignocellulosic feedstock into fuels. Lignin one of the components of secondary cell wall tightly holds the polysaccharides thereby enhancing the recalcitrance and complexity in the biomass. Laccases (LAC) and peroxidases (PRX) are the major phenyl-oxidases playing key functions during the polymerization of monolignols into lignin. Yet, the functions of laccase and peroxidases gene families remained largely unknown. Hence, the objective of this conducted study is to understand the role of specific LAC and PRX in Populus wood formation and to further investigate how the altered Lac and Prx expression affects biomass recalcitrance and plant growth. This study of heterologous expression of Arabidopsis Lac and Prx genes was conducted in poplar to avoid any otherwise occurring co-suppression mechanism during the homologous overexpression of highly expressed native genes. In the pursuit of optimizing lignocellulosic biomass for biofuel production, the present study focuses on harnessing the enzymatic potential of Arabidopsis thaliana Laccase2, Laccase4, and Peroxidase52 through heterologous expression. We overexpressed selected Arabidopsis laccase2 (AtLac2), laccase4 (AtLac4), and peroxidase52 (AtPrx52) genes, based on their high transcript expression respective to the differentiating xylem tissues in the stem, in hybrid poplar (cv. 717) expressed under the developing xylem tissue-specific promoter, DX15 characterized the transgenic populus for the investigation of growth phenotypes and recalcitrance efficiency. Bioinformatics analyses conducted on AtLac2 and AtLac4 and AtPrx52, revealed the evolutionary relationship between the laccase gene and peroxidase gene homologs, respectively. Transgenic poplar plant lines overexpressing the AtLac2 gene (AtLac2-OE) showed an increase in plant height without a change in biomass yield as compared to the controls; whereas AtLac4-OE and AtPrx52-OE transgenic lines did not show any such observable growth phenotypes compared to their respective controls. The changes in the levels of lignin content and S/G ratios in the transgenic poplar resulted in a significant increase in the saccharification efficiency as compared to the control plants. Overall, saccharification efficiency was increased by 35–50%, 21–42%, and 8–39% in AtLac2-OE, AtLac4-OE, and AtPrx52-OE transgenic poplar lines, respectively, as compared to their controls. Moreover, the bioengineered plants maintained normal growth and development, underscoring the feasibility of this approach for biomass improvement without compromising overall plant fitness. This study also sheds light on the potential of exploiting regulatory elements of DX15 to drive targeted expression of lignin-modifying enzymes, thereby providing a promising avenue for tailoring biomass for improved biofuel production. These findings contribute to the growing body of knowledge in synthetic biology and plant biotechnology, offering a sustainable solution to address the challenges associated with lignocellulosic biomass recalcitrance.},
doi = {10.1186/s13068-023-02452-7},
journal = {Biotechnology for Biofuels and Bioproducts},
number = 1,
volume = 17,
place = {United States},
year = {Sat Jan 13 00:00:00 EST 2024},
month = {Sat Jan 13 00:00:00 EST 2024}
}
Works referenced in this record:
Lignin Biosynthesis
journal, June 2003
- Boerjan, Wout; Ralph, John; Baucher, Marie
- Annual Review of Plant Biology, Vol. 54, Issue 1, p. 519-546
Glycine‐rich RNA‐binding protein7 affects abiotic stress responses by regulating stomata opening and closing in Arabidopsis thaliana
journal, July 2008
- Kim, Jin Sun; Jung, Hyun Ju; Lee, Hwa Jung
- The Plant Journal, Vol. 55, Issue 3
The laccase multigene family in Arabidopsis thaliana: towards addressing the mystery of their gene function(s)
journal, November 2010
- Turlapati, Phanikanth V.; Kim, Kye-Won; Davin, Laurence B.
- Planta, Vol. 233, Issue 3
Analysis and expression of the class III peroxidase large gene family in Arabidopsis thaliana
journal, April 2002
- Tognolli, Michael; Penel, Claude; Greppin, Hubert
- Gene, Vol. 288, Issue 1-2
Subcellular Relocalization and Positive Selection Play Key Roles in the Retention of Duplicate Genes ofPopulusClass III Peroxidase Family
journal, June 2014
- Ren, Lin-Ling; Liu, Yan-Jing; Liu, Hai-Jing
- The Plant Cell, Vol. 26, Issue 6
The class III peroxidase multigenic family in rice and its evolution in land plants☆☆☆
journal, July 2004
- Passardi, Filippo; Longet, David; Penel, Claude
- Phytochemistry, Vol. 65, Issue 13
Oxidation of hydroxycinnamic acid and hydroxycinnamyl alcohol derivatives by laccase and peroxidase. Interactions among p -hydroxyphenyl, guaiacyl and syringyl groups during the oxidation reactions
journal, January 1995
- Takahama, Umeo
- Physiologia Plantarum, Vol. 93, Issue 1
A lignin-specific peroxidase in tobacco whose antisense suppression leads to vascular tissue modification
journal, September 2003
- Blee, Kristopher A.; Choi, Joon W.; O'Connell, Ann P.
- Phytochemistry, Vol. 64, Issue 1
Differential Activity and Structure of Highly Similar Peroxidases. Spectroscopic, Crystallographic, and Enzymatic Analyses of Lignifying Arabidopsis thaliana Peroxidase A2 and Horseradish Peroxidase A2 † , ‡
journal, September 2001
- Nielsen, Kåre Lehmann; Indiani, Chiara; Henriksen, Anette
- Biochemistry, Vol. 40, Issue 37
Isolation and Characterization of a Novel Peroxidase Gene ZPO-C Whose Expression and Function are Closely Associated with Lignification during Tracheary Element Differentiation
journal, April 2006
- Sato, Yasushi; Demura, Taku; Yamawaki, Ken
- Plant and Cell Physiology, Vol. 47, Issue 4
PeroxiBase: A class III plant peroxidase database
journal, March 2006
- Bakalovic, Nenad; Passardi, Filippo; Ioannidis, Vassilios
- Phytochemistry, Vol. 67, Issue 6
Progress of Lignification Mediated by Intercellular Transportation of Monolignols During Tracheary Element Differentiation of Isolated Zinnia Mesophyll Cells
journal, September 2001
- Hosokawa, Mami; Suzuki, Shiro; Umezawa, Toshiaki
- Plant and Cell Physiology, Vol. 42, Issue 9
Apoplastic Peroxidases and Lignification in Needles of Norway Spruce (Picea abies L.)
journal, September 1994
- Polle, A.; Otter, T.; Seifert, F.
- Plant Physiology, Vol. 106, Issue 1
The Genome of Black Cottonwood, Populus trichocarpa (Torr. & Gray)
journal, September 2006
- Tuskan, G. A.; DiFazio, S.; Jansson, S.
- Science, Vol. 313, Issue 5793, p. 1596-1604
Zinnia elegans uses the same peroxidase isoenzyme complement for cell wall lignification in both single-cell tracheary elements and xylem vessels
journal, January 2004
- Lopez-Serrano, M.; Fernandez, M. D.; Pomar, F.
- Journal of Experimental Botany, Vol. 55, Issue 396
Down-regulation of an anionic peroxidase in transgenic aspen and its effect on lignin characteristics
journal, June 2003
- Li, Yahong; Kajita, Shinya; Kawai, Shinya
- Journal of Plant Research, Vol. 116, Issue 3
Crystal structure of the type-2 Cu depleted laccase from Coprinus dnereus at 2.2 Å resolution
journal, April 1998
- Ducros, Valérie; Brzozowski, Andrzej Marek; Wilson, Keith S.
- Nature Structural Biology, Vol. 5, Issue 4
Gene discovery in the wood-forming tissues of poplar: Analysis of 5,692 expressed sequence tags
journal, October 1998
- Sterky, F.; Regan, S.; Karlsson, J.
- Proceedings of the National Academy of Sciences, Vol. 95, Issue 22
Gene structure and molecular analysis of the laccase-like multicopper oxidase (LMCO) gene family in Arabidopsis thaliana
journal, June 2005
- McCaig, Bonnie C.; Meagher, Richard B.; Dean, Jeffrey F. D.
- Planta, Vol. 221, Issue 5
Laccases and Peroxidases Co-Localize in Lignified Secondary Cell Walls throughout Stem Development
journal, July 2020
- Hoffmann, Natalie; Benske, Anika; Betz, Heather
- Plant Physiology, Vol. 184, Issue 2
Inactivation of LACCASE8 and LACCASE5 genes in Brachypodium distachyon leads to severe decrease in lignin content and high increase in saccharification yield without impacting plant integrity
journal, July 2019
- Le Bris, Philippe; Wang, Yin; Barbereau, Clément
- Biotechnology for Biofuels, Vol. 12, Issue 1
Lignin engineering through laccase modification: a promising field for energy plant improvement
journal, September 2015
- Wang, Jinhui; Feng, Juanjuan; Jia, Weitao
- Biotechnology for Biofuels, Vol. 8, Issue 1
Genetic Modification of KNAT7 Transcription Factor Expression Enhances Saccharification and Reduces Recalcitrance of Woody Biomass in Poplars
journal, October 2021
- Ahlawat, Yogesh Kumar; Nookaraju, Akula; Harman-Ware, Anne E.
- Frontiers in Plant Science, Vol. 12
Biomass Recalcitrance: Engineering Plants and Enzymes for Biofuels Production
journal, February 2007
- Himmel, M. E.; Ding, S.-Y.; Johnson, D. K.
- Science, Vol. 315, Issue 5813, p. 804-807
Significance of Lignin S/G Ratio in Biomass Recalcitrance of Populus trichocarpa Variants for Bioethanol Production
journal, December 2017
- Yoo, Chang Geun; Dumitrache, Alexandru; Muchero, Wellington
- ACS Sustainable Chemistry & Engineering, Vol. 6, Issue 2
Lignification in Trees: Indication of Exclusive Peroxidase Participation
journal, April 1973
- Harkin, J. M.; Obst, J. R.
- Science, Vol. 180, Issue 4083
Laccase and the Deposition of Lignin in Vascular Plants
journal, January 1994
- Dean, Jeffrey F. D.; Eriksson, Karl-Erik L.
- hfsg, Vol. 48, Issue s1
Catalytic Profile of Arabidopsis Peroxidases, AtPrx-2, 25 and 71, Contributing to Stem Lignification
journal, August 2014
- Shigeto, Jun; Nagano, Mariko; Fujita, Koki
- PLoS ONE, Vol. 9, Issue 8
LACCASE Is Necessary and Nonredundant with PEROXIDASE for Lignin Polymerization during Vascular Development in Arabidopsis
journal, October 2013
- Zhao, Qiao; Nakashima, Jin; Chen, Fang
- The Plant Cell, Vol. 25, Issue 10
Laccase Down-Regulation Causes Alterations in Phenolic Metabolism and Cell Wall Structure in Poplar
journal, April 2002
- Ranocha, Philippe; Chabannes, Matthieu; Chamayou, Simon
- Plant Physiology, Vol. 129, Issue 1
Knockdown of a laccase in Populus deltoides confers altered cell wall chemistry and increased sugar release
journal, April 2016
- Bryan, Anthony C.; Jawdy, Sara; Gunter, Lee
- Plant Biotechnology Journal, Vol. 14, Issue 10
Peroxidase Activity in Poplar Inoculated with Compatible and Incompetent Isolates of Paxillus involutus
journal, June 2007
- Gafur, Abdul; Schutzendubel, Andres; Polle, Andrea
- HAYATI Journal of Biosciences, Vol. 14, Issue 2
Disruption of LACCASE4 and 17 Results in Tissue-Specific Alterations to Lignification of Arabidopsis thaliana Stems
journal, March 2011
- Berthet, Serge; Demont-Caulet, Nathalie; Pollet, Brigitte
- The Plant Cell, Vol. 23, Issue 3
Ptr-miR397a is a negative regulator of laccase genes affecting lignin content in Populus trichocarpa
journal, June 2013
- Lu, S.; Li, Q.; Wei, H.
- Proceedings of the National Academy of Sciences, Vol. 110, Issue 26
Sugar release and growth of biofuel crops are improved by downregulation of pectin biosynthesis
journal, February 2018
- Biswal, Ajaya K.; Atmodjo, Melani A.; Li, Mi
- Nature Biotechnology, Vol. 36, Issue 3
The MEME Suite
journal, May 2015
- Bailey, Timothy L.; Johnson, James; Grant, Charles E.
- Nucleic Acids Research, Vol. 43, Issue W1
New and continuing developments at PROSITE
journal, November 2012
- Sigrist, Christian J. A.; de Castro, Edouard; Cerutti, Lorenzo
- Nucleic Acids Research, Vol. 41, Issue D1
Cytoscape StringApp: Network Analysis and Visualization of Proteomics Data
journal, November 2018
- Doncheva, Nadezhda T.; Morris, John H.; Gorodkin, Jan
- Journal of Proteome Research, Vol. 18, Issue 2
The STRING database in 2023: protein–protein association networks and functional enrichment analyses for any sequenced genome of interest
journal, November 2022
- Szklarczyk, Damian; Kirsch, Rebecca; Koutrouli, Mikaela
- Nucleic Acids Research, Vol. 51, Issue D1
ClueGO: a Cytoscape plug-in to decipher functionally grouped gene ontology and pathway annotation networks
journal, February 2009
- Bindea, Gabriela; Mlecnik, Bernhard; Hackl, Hubert
- Bioinformatics, Vol. 25, Issue 8