1. A self-assembling evolutionary marvel by: Drew Sowersby Supramolecular Chemistry 5390 Dr. Wendi David

2. Cellulose is the most abundant source of bioenergy on earth Rubin, E. Nature, 2008, 454, 841-845. BIOFUELS Applications

3. Singh, S.; Simmons, B. a; Vogel, K. P. Biotechnology and bioengineering. 2009, 104, 68-75. The break down of cellulose involves 2 major steps Pretreatment mechanical (shear) chemical (ionic, dilute acids) thermochemical (heat) Hydrolysis enzymatic ( cellulases ) chemical (strong acids) switchgrass – before and after ionic liquid pretreatment

4. Graphic: Wikipedia Hydrolysis to glucose is achieved via 3 enzymatic steps

5. http://www.wzw.tum.de/mbiotec/celostruct.htm The Cellulosome Concept  Bayer and Lamed (1983)  Clostridium thermocellum most studied  extracellular molecular machine  molecular weight of over 2 MDa (2,000,000 Da) Lamed, R. et al. Biotechnol. Bioeng. Symp., 1983, 13, 163–18.

6. Characterization of the cellulosome Complex crystallization Isothermal calorimetry Protein purification Recombinant DNA technology Bioinformatics and modeling Electron microscopy Surface plasmon resonance Binding Structure Composition

7. What does the research say about the cellulosome? 1.Shoham, Y.; Lamed, R.; Bayer, E.A. Trends Microbiol., 1999, 7, 275-281. 2.Schwarz, W. H. Appl. Microbiol. Biotechnol. 2001, 56, 634–649. 3.Pagès, S. et al. Proteins, 1997, 29, 517–527. 4.Carvahlo, A. L. et al. PNAS, 2003, 100, 13809–13814. Mechanism Cellulosome self-assembly driven by non-specific but highly complementary cohesin/dockerin interactions Features o contains18 different dockerin containing enzymes cellulases ranging in molecular weights of 40-180 kDa dockerin is a 23-residue tandemly repeated sequence dockerin contains 2 calcium binding loop-helix motifs o cooperative binding by carbohydrate binding modules (CBMs) o binding between dockerin and cohesin is species specific o cohesin is a 9-stranded β - sandwich with a jellyroll topology o Scaffoldin-cellulase superstructure results in synergistic cellulolytic activity not seen with free floating cellulases

8. Scaffoldin and the dockerin-cohesin complex scaffoldin

9. Polycellulosomal superstructures  extracellular construction  organismal monstrosity  resting vs. protracted “Plasticity theory” 1. Shoham, Y.; Lamed, R.; Bayer, E.A. Trends Microbiol., 1999, 7, 275-281. 2. Bomble, Y. J. et al. J. Biol. Chem., 2011, 286, 5614-5623.

10. Video Simulation wrap-up Bomble, Y. J. et al. J. Biol. Chem., 2011, 286, 5614-5623.