Reducing biomass recalcitrance by heterologous expression of a bacterial peroxidase in tobacco (Nicotiana benthamiana)

Ayalew Ligaba-Osena, Bertrand Hankoua, Kay Dimarco, Robert Pace, Mark Crocker, Jesse McAtee, Nivedita Nagachar, Ming Tien, Tom L. Richard

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Commercial scale production of biofuels from lignocellulosic feed stocks has been hampered by the resistance of plant cell walls to enzymatic conversion, primarily owing to lignin. This study investigated whether DypB, the lignin-degrading peroxidase from Rodococcus jostii, depolymerizes lignin and reduces recalcitrance in transgenic tobacco (Nicotiana benthamiana). The protein was targeted to the cytosol or the ER using ER-targeting and retention signal peptides. For each construct, five independent transgenic lines were characterized phenotypically and genotypically. Our findings reveal that expression of DypB in the cytosol and ER does not affect plant development. ER-targeting increased protein accumulation, and extracts from transgenic leaves showed higher activity on classic peroxidase substrates than the control. Intriguingly, in situ DypB activation and subsequent saccharification released nearly 200% more fermentable sugars from transgenic lines than controls, which were not explained by variation in initial structural and non-structural carbohydrates and lignin content. Pyrolysis-GC-MS analysis showed more reduction in the level of lignin associated pyrolysates in the transgenic lines than the control primarily when the enzyme is activated prior to pyrolysis, consistent with increased lignin degradation and improved saccharification. The findings reveal for the first time that accumulation and in situ activation of a peroxidase improves biomass digestibility.

Original languageEnglish (US)
Article number17104
JournalScientific reports
Volume7
Issue number1
DOIs
StatePublished - Dec 1 2017

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Lignin
Biomass
Peroxidase
Tobacco
Cytosol
Plant Development
Biofuels
Plant Cells
Protein Transport
Protein Sorting Signals
Cell Wall
Carbohydrates
Enzymes
Proteins

All Science Journal Classification (ASJC) codes

  • General

Cite this

Ligaba-Osena, Ayalew ; Hankoua, Bertrand ; Dimarco, Kay ; Pace, Robert ; Crocker, Mark ; McAtee, Jesse ; Nagachar, Nivedita ; Tien, Ming ; Richard, Tom L. / Reducing biomass recalcitrance by heterologous expression of a bacterial peroxidase in tobacco (Nicotiana benthamiana). In: Scientific reports. 2017 ; Vol. 7, No. 1.
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abstract = "Commercial scale production of biofuels from lignocellulosic feed stocks has been hampered by the resistance of plant cell walls to enzymatic conversion, primarily owing to lignin. This study investigated whether DypB, the lignin-degrading peroxidase from Rodococcus jostii, depolymerizes lignin and reduces recalcitrance in transgenic tobacco (Nicotiana benthamiana). The protein was targeted to the cytosol or the ER using ER-targeting and retention signal peptides. For each construct, five independent transgenic lines were characterized phenotypically and genotypically. Our findings reveal that expression of DypB in the cytosol and ER does not affect plant development. ER-targeting increased protein accumulation, and extracts from transgenic leaves showed higher activity on classic peroxidase substrates than the control. Intriguingly, in situ DypB activation and subsequent saccharification released nearly 200{\%} more fermentable sugars from transgenic lines than controls, which were not explained by variation in initial structural and non-structural carbohydrates and lignin content. Pyrolysis-GC-MS analysis showed more reduction in the level of lignin associated pyrolysates in the transgenic lines than the control primarily when the enzyme is activated prior to pyrolysis, consistent with increased lignin degradation and improved saccharification. The findings reveal for the first time that accumulation and in situ activation of a peroxidase improves biomass digestibility.",
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Reducing biomass recalcitrance by heterologous expression of a bacterial peroxidase in tobacco (Nicotiana benthamiana). / Ligaba-Osena, Ayalew; Hankoua, Bertrand; Dimarco, Kay; Pace, Robert; Crocker, Mark; McAtee, Jesse; Nagachar, Nivedita; Tien, Ming; Richard, Tom L.

In: Scientific reports, Vol. 7, No. 1, 17104, 01.12.2017.

Research output: Contribution to journalArticle

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AU - Ligaba-Osena, Ayalew

AU - Hankoua, Bertrand

AU - Dimarco, Kay

AU - Pace, Robert

AU - Crocker, Mark

AU - McAtee, Jesse

AU - Nagachar, Nivedita

AU - Tien, Ming

AU - Richard, Tom L.

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