Plant Biotechnology for Biofuels

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1 Plant Biotechnology for Biofuels Markus Energy Biosciences Institute Department of Plant and Microbial Biology UC Berkeley Princeton University, October 14, 2013

2 CO 2 -concentration in the atmosphere! Annual Cycle! 6 ppm! 11 May, 2013: 400 ppm! Jan Apr Jul Oct Jan! 280 ppm (Preindustrial)! Today! ppm! 185 ppm (Ice ages)! 800,000! 600,000! 400,000! 200,000! Today! Scripps Institute of Oceanography; National Climatic Data Center!

3 Carbon fixation in plants CO 2 H 2 O Chloroplast Triose-P Chloroplast Starch (storage) Triose-P Hexose-P Cell Wall Polymers (structure) CO 2 H 2 O Glycolysis (energy) Sucrose (transport)

4 Plant cell walls and Carbon Cycle Assimilated CO 2 Plant Biomass (land, worldwide) Cell walls (land, worldwide) 55 x10 9 t / a x 10 9 t / a x 10 9 t / a Field et al., 1998; Porter and Vilar, 1997; and Keegstra 2008 Annual global production of chemicals x 10 9 t (without pharmaceuticals) BASF report 2010 Trends in the chemical industry Cell walls represent the dominant biological carbon sequestration system

5 The Plant Cell Wall Cellulose Hemicellulose Pectin xyloglucan galactomannan arabinoxylan Homogalacturonan Ca 2+ -crosslinked non-methylesterified methylesterified RG I (galactan) (arabinan) Rhamnogalacturonan RG II (boron-diester)

6 The mature Plant Cell Wall Cellulose Hemicellulose Polyphenols (Lignin)

7 Function of plant cell walls - structural integrity of cell/ tissue/ plant - cell shape/ plant morphology - cell adhesion - barrier (mechanical/ path. defense) Redwoods: up to 379 feet high, up to 2200 y old - dynamic entity of the cell - cell expansion Buchanan, 1999 Keith Roberts - reposit for signaling molecules - pathogen attack

8 The Plant Cell Wall: Diversity plant species plant tissue plant cell Courtesy of Paul Knox, Univeristy of Leeds

9 Assessment of plant cell wall structural diversity: Whole lignocellulosic 2D-NMR Plant tissue (Lignocellulosics) Ball milling Dissolution in organic solvent 2D-NMR spectroscopy 10 mg (dry) DMSO-d 6 /[Emim]OAc-d µl : 10 µl Kun Cheng

10 Whole lignocellulosic 2D-NMR 1 H- 13 C HSQC Kun Cheng

11 Whole lignocellulosic 2D-NMR Anomeric polysaccharides

12 Whole lignocellulosic 2D-NMR Aromatic Lignin

13 Whole lignocellulosic 2D-NMR Aliphatic Lignin linkages

14 Whole lignocellulosic 2D-NMR Glc Xyl Ara Man GlcA Ac-xylan Molar percentage (Miscanthus) > /- 3 +/- 1 +/ / /- 2 Guaiacyl (G) Syringyl (S) P-hydroxyphenyl (H) S/G ratio >1.8 +/- 1 +/- 1 +/- 0.5 Aryl-ether (A) Phenylcoumaran (B) Resinol (C) Dibenzodioxacin (D) /- 3 +/ /- 1 Cheng, Sorek, Zimmermann, Wemmer, (2013) Analytical chemistry

15 Structural Space of lignocellulosics Plant species Model plant Arabidopsis Energy Grass Switchgrass Miscanthus Agriculture residues Rice Sorghum Sugarcane Maize Forestry resources Pine Maple Willow Poplar Eucalyptus

16 Lignocellulosic structural diversity GlcA S G β-o-4 β-5 H Ara Non- Cell Lignin β-β 5-5 β-o-4 pca FA Agave americana Agave fourcroydes Agave tequilina Cellulose Poplar wood Willow wood Eucalyptus wood Pine wood Sugarcane bagasse Cordgrass straw Miscanthus straw Sorghum Maize leaf Maize cob

17 Structural Space of lignocellulosics Principal component clustering Pine (17%) Eucalyptus Arabidopsis Switchgrass Maize Sugarcane Miscanthus Willow Maple Sorghum Poplar (35%) Plant species Grasses Wood (Hardwood Softwood) Kun Cheng

18 Alternative to fossil fuels: Plant biomass CO 2 oil vehicle plants biorefinery CO 2 refinery vehicle gasoline biofuel Plants capture energy from the sun Plants convert energy into durable chemicals Plants sequester CO 2 from the atmosphere

19 Lignocellulosics as renewable resource Cell walls (lignocellulosics) are non-food components Lignocellulosics are highly abundant Dominant biological carbon sequestration system The conversion of lignocellulosics is energy and costintensive

20 Biomass to Biofuels Biomass Vegetable Oil Chemical Conversion steps Transesterification Fuel Biodiesel Sugar Starch Pretreatment Lignocellulosics Biological Fermentation Enzymatic hydrolysis Thermochemical Syngas production Pyrolysis Catalytic Conversion Fischer Tropsch process Ethanol Butanol Hydrocarbons Biological Anaerobic gasification Methane

21 Challenges: Plant biology 2 nd Gen Biofuels Increase plant biomass/ ha Choice of crops Increase resource efficiency (water, fertilizer) Delayed flowering (energy maize: Increase the yield of fermentable sugars Create a more open wall architecture (reduce, alter lignin) Reduce process inhibitors Increase cellulose (amorphous), hemicelluloses (hexoses) Van Holme, 2007

22 Acknowledgements lab Lifeng Liu Jonathan Paulitz Guangyan Xiong Amancio Souza Nasim Mansoori Alex Schultink Julia Schulbert Dan Naylor Collaborators: Sarah Hake, USDA Albany Patrick Brown, UIUC ISU transformation facility

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