Biomass conversion: opportunities in lignin management · 2016. 12. 6. · Biomass conversion:...
Transcript of Biomass conversion: opportunities in lignin management · 2016. 12. 6. · Biomass conversion:...
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Biomass conversion: opportunities in lignin management
Clint ChappleDepartment of Biochemistry
Purdue University
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Targets for biomass improvement
• Yield
• Water and nutrient use efficiency
• Pest and pathogen resistance
• Biomass quality
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Targets for biomass improvement
• Yield
• Water and nutrient use efficiency
• Pest and pathogen resistance
• Biomass quality
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Lignin is critical for plant survival
• structural support• water transport
• important impact on bioenergy and bioprocessing
• large commitment of fixed carbon
• high energy density
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Lignin is a biosynthetically plastic polymer
Bonawitz and Chapple, Ann Rev Genetics, 2010
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Lignin is a biosynthetically plastic polymer
OOMe
OMeO OH
O
OH
OMe
O
HO
OMe
OH
OMeO
HO
O
HO
HO
OH
O
OH
OMe
OMe
O
MeO
OH
OMe
HO OH
HO
OH
MeO
• Nucleic acid synthesis• Template-dependent
• Protein synthesis• Template-dependent
• Polysaccharide synthesis• Enzyme specificity-directed
• Lignin synthesis• Random radical coupling dependent on precursor supply
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Opportunities for lignin engineering
• Block lignin synthesis
• Change lignin composition
• Redirect lignin monomers
• Alter lignin localization
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Mutant screens can identify genes important to processes of interest
Normal plant
↓ mutagenesis
Mutant with trait of interest
↓ gene identification
Understanding of gene function
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Lignin is a biosynthetically plastic polymer
Bonawitz and Chapple, Ann Rev Genetics, 2010
![Page 10: Biomass conversion: opportunities in lignin management · 2016. 12. 6. · Biomass conversion: opportunities in lignin management Clint Chapple Department of Biochemistry Purdue University](https://reader036.fdocuments.in/reader036/viewer/2022081621/61249da2c9029e4c2f523fc3/html5/thumbnails/10.jpg)
Lignin modification-induced dwarfing (LMID)
Schilmiller et al., Plant J. 2009
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Lignin is a biosynthetically plastic polymer
Bonawitz and Chapple, Ann Rev Genetics, 2010
![Page 12: Biomass conversion: opportunities in lignin management · 2016. 12. 6. · Biomass conversion: opportunities in lignin management Clint Chapple Department of Biochemistry Purdue University](https://reader036.fdocuments.in/reader036/viewer/2022081621/61249da2c9029e4c2f523fc3/html5/thumbnails/12.jpg)
Lack of C3'H reduces lignin content andleads to novel lignins
Franke et al., Plant J, 2002
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Lignin is a biosynthetically plastic polymer
Bonawitz and Chapple, Ann Rev Genetics, 2010
![Page 14: Biomass conversion: opportunities in lignin management · 2016. 12. 6. · Biomass conversion: opportunities in lignin management Clint Chapple Department of Biochemistry Purdue University](https://reader036.fdocuments.in/reader036/viewer/2022081621/61249da2c9029e4c2f523fc3/html5/thumbnails/14.jpg)
ref4 and rfr1 are required for lignin regulation
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ref4 and rfr1 are required for lignin regulation
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REF4 and RFR1 are components of theMediator complex
P Cramer Lab, Gene Center, University of Munich
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med5a/bref8-1
med5a/bref8-2
wild type ref8-1med5a/b ref8-2
Disruption of MED5 rescues the dwarf phenotype of ref8
Bonawitz et al., Nature, 2014
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ref8-1 mutants show widespread MED5-dependent transcriptional reprogramming
med5a/b89
216med5a/b ref8-1
ref8-14441
95
39
43025
med5a/b19
196med5a/b ref8-1
ref8-13360
54
12
37147
up-regulated down-regulated
(DeSeq, false discovery rate <0.05)
Bonawitz et al., Nature, 2014
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med5a/b ref8-1 mutants synthesize predominantly H lignin
syringyl (S)
guaiacyl (G)
p-hydroxyphenyl (H)
wild type med5a/b med5a/b ref8-1
Yuki Tobimatsu, John Ralph, Wisconsin
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med5a/b ref8 mutants show facilitated saccharification
0
20
40
60
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0 24 48 72
0
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0 24 48 72
gluc
ose
rele
ase
(%)
wild type
med5a/b
med5a/b ref8-1
time (hrs)
no pretreatment liquid hot water pretreated
Eduardo Ximenes, Mike Ladisch, Purdue
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Suppressor screens can identify genes important to processes of interest
Normal plant
↓ mutagenesis
Mutant with trait of interest
↓ mutagenesis
Mutant plant in which trait is abolished
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Suppressors of ref4-3 may identify other proteins required for phenylpropanoid regulation
Dolan et al., unpublished
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Whole genome sequencing identified intragenic suppressor mutations and Mediator subunit suppressors
Dolan et al., unpublished
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REF4 and RFR1 are components of theMediator complex
P Cramer Lab, Gene Center, University of Munich
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Maybe lignin isn’t all bad
• Significant proportion of plant biomass
• More highly reduced than sugars and polysaccharides
• Amenable to catalytic degradation and conversion
OOMe
OMeO OH
O
OH
OMe
O
HO
OMe
OH
OMeO
HO
O
HO
HO
OH
O
OH
OMe
OMe
O
MeO
OH
OMe
HO OH
HO
OH
MeO
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Maybe lignin isn’t all bad
Parsell et al., Green Chemistry 2015
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Maybe lignin isn’t all bad
Shuai et al., Science 2016
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Summary
• We are developing an understanding of the catalysts and regulators that provide us with control over lignin
• The study of Arabidopsis has revealed a previously unknown regulatory circuit for lignin biosynthesis that involves Mediator
• The dwarfing seen in some lignin-deficient plants can be suppressed genetically, indicating that it is an active protein-mediated process
• Plants with high H-lignin are viable and could provide novel inputs for the biorefinery
• Catalytic conversion of lignin into fuels may make more efficient use of biomass
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Acknowledgements
Purdue and C3Bio
Nick BonawitzWhitney Dolan
Jake StoutCandy Mao
Bryon DonohoePeter Ciesielski
Eduardo XimenesMike Ladisch
Brian DilkesCharles Addo-Quaye
GCEP
Sirius Li
Yuki TobimatsuJohn Ralph
Chris McClellanClaire Halpin
Wout Boerjan