Utilizing Lignin for Biofuel Production

1. Utilizing Lignin for Biofuel Production Zach Krehlik

2. Overview • Biofuel • Components of Cells • Lignin • White Rot Fungi

3. Biofuel Fermentation • Conversion of biological matter to combustible fuels • Two Major Reactions • Hydrolysis • Biomass is converted to glucose • Fermentation • C6H12O6 2 CO2 + 2CH3CH2OH • Yeast fermentation produces 15% ethanol, which can be purified to 95%

4. Biofuel Sources Corn Soybeans Coconut oil Jatropha Catfish oil Cheese scraps Algae Most of these are not very realistic sources

5. Biofuel Sources Industrial Products • Utilizes pulp, wood chips, and other otherwise useless by products • Requires time consuming intermediate steps

6. Cell Wall Composition • Cellulose 35-50% • Xylan 20-35% • Lignin 10-25% • Proteins 1-5%

7. Lignin • Complex structure • Not a polymer made of one simple monomer • More similar to proteins • Variable structure • If not utilized, a significant amount of cellular carbon is wasted

8. Lignin + ???? 

9. Lignin Modification • Acid Treatment • Time consuming and not very cost efficient • White Rot Fungi • One of the few organisms known to process lignin • Multiple enzymes have been isolated • Transgenic yeasts • Combination

10. White Rot Fungi • White Rot Fungi • Ceriporiopsissubvermispora, Dichomitussqualens, Pleurotusostreatus, Coriolusversicolor • 3 methods of lignin degradation by white rot fungi • Lignin Peroxidase • Highly reactive • Manganese Peroxidase • Highly reactive • Laccase • Initiates depolarization • Low substrate specificity allows it to break down the highly variable molecule

11. White Rot Fungi • Acid Pretreatments take up to 6 weeks • White Rot converts it in a few days • The wood chips are then separated from the fungi and exposed to the fermentation yeasts • Wood chips treated with White Rot have yielded up to 160% the ethanol as the chips not pretreated

12. White Rot Fungi Treatment Weight and component loss of beech wood chips after exposure to White Rot Fungi. Lignin degradation was increased by over 20%

13. White Rot Fungi Treatment • A cooperative study across multiple universities in Japan have been using transgenic yeast strains to efficiently break down cellular components. • Their actual lignin conversion is 93% of the theoretical!

14. The Future of Lignin Degradation • Transgenic yeasts combined with a less time consuming chemical process will likely be utilized commonly on a large scale

15. Questions?

16. Sources • Biomass Recalcitrance: Engineering Plants and Enzymes for Biofuels Production. Michael E. Himmel, Shi-You Ding, David K. Johnson, William S. Adney, Mark R. Nimlos, John W. Brady, Thomas D. Foust. Science 9 February 2007: Vol. 315 no. 5813 pp. 804-807. • Anaerobic fermentation of glycerol: a path to economic viability for the biofuels industry. Syed Shams Yazdani and Ramon Gonzalez. Department of Chemical and Biomolecular Engineering, Rice University, 6100 Main Street, MS-362, Houston, TX 77005, USA. Energy biotechnology / Environmental biotechnology Volume 18, Issue 3, June 2007, pp. 213-219. • Challenges in Engineering Microbes for Biofuels Production. Gregory Stephanopoulos. Science 9 February 2007: Vol. 314 no. 5813 pp. 801-804. • Lignin modification improves fermentable sugar yields for biofuel production. Fang Chen and Richard A Dixon. Nature Biotechnology 25, 759-761. June 17, 2007. • Emerging strategies of lignin engineering and degradation for cellulosic biofuel production. Jing-KeWeng, Xu Li, Nicholar D. Bonowitz, and Clint Chapple. Department of Biochemistry, Purdue University, 175 South University Sreet, West Lafayette, IN 47907-2063, United States. April 9, 2008. • Bioorganosolve pretreatments for simultaneous saccharification and fermentation of beech wood by ethanolysis and white rot fungi. HiromichiItoh, Masanori Wada, Yoichi Honda, Masaaki Kuwahara, and Takashi Watanabe. Laboratory of Biomass Conversion, Wood Research Institute, Kyoto University, Gokasho, Uji, Kyoto, Japan. July 11, 2003. • Ethanol Production from Agricultural Biomass Substrates. Rodney J. Bothast and Badal C. Saha. Fermentation Biochemistry Research Unit National Center for Agricultural Utilization Research USDA Agricultural Research Service Peoria, Illinois 61604. April 15, 2008. • Direct and Efficient Production of Ethanol from Cellulosic Material with a Yeast Strain Displaying Cellulolytic Enzymes. Applied and Environmental Microbiology, October 2002, pp. 5136-5141, Vol 68, No. 10. Yasuya Fujita, Shouji Takahashi, Mitsuyoshi Ueda, Atsuo Tanaka, Hirofumi Okada, Yasushi Morikawa, Takashi Kawaguchi, Motoo Arai, Hideki Fukuda, and Akihiko Kondo. Division of Molecular Sciences, Graduate School of Science and Technology of Kobe University, Department of Chemical Science and Engineering, Faculty of Engineering, Kobe Univiersity, Department of Synthetic Chemistyr and Biological Chemistry, Graduate School of Engineering, Kyoto University, Yoshida, Sakyo-ku, Kyoto 606-8501, Department of Bioengineering, Nagaoka University of Technology, 1603-1 Kamitomioka, Nagaoka, Niigata 940-2188, Department of Agricultural Chemistry, College of Agriculture, University of Osaka Prefecture, Sakai, Osaka 599-8531, Japan. • Ethanol from lignocellulosic biomass: technoeconomic performance in short-, middle- and long-term. Biomass and Bioenergy, Vol. 28, Issue 4, April 2005, pp. 384-410. Carlo N. Hamelinck, Geertje van Hooijdonk, and Andre PC Faaij. • Cellodextrin Transport in Yeast for Improved Biofuel Production. Jonathon M. Galazka et al. Science 330, 84 (2010). • Science Creative Quarterly. Issue 6, year 2011. http://www.scq.ubc.ca/food-microbiology-the-basics-and-the-details-of-cheese-production/