1. Enzymatic Hydrolysis of cellulose for production of fuel ethanol by SSF Presented by Alaaedine Talab

2. What is SSF?

3. The reaction

4. The reaction continued β(1→ 4)

5. Purpose of SSF Minimize Hydrolysis byproducts Economically favorable Minimize inhibitory factors Eliminate transfer time

6. SSF Kinetics Cellulose cellobiose glucose r1r1 r2r2

7. Experimental Procedure Effects of specific components of SSF system Assays done in Y/P medium (10g/L yeast extract, 20g/L peptone), PH = 5.0 at 38C Trichoderma Reesei Initial rates determined after enzyme deactivation(20 minutes boiling water, denatur) 15s intervals 1 minute duration

8. How do we measure enzyme activity as an uncoupled system? 3g/L

9. We can simplify the volumetric rate constants = constant

10. Effect of cellobiose on cellulase activity

11. Avg. Effect of cellobiose on B-glucosidase K 2B ?

12. Effect of cellobiose on B-glucosidase Maximal rate

13. Glucose concentration and enzyme activity

14. Ethanol concentration and Enzyme activity K 2E = NEGLIGIBLE

15. Summary

16. Putting it all together

17. Putting it all together continued Almost perfect fit but not yet Why?

18. First order rate loss added to model Fail

19. (Conclusion)So what? R^2> 0.98 Developed a quantitative mathematical model that can predict SSF progress This applied to enzymes from T. risea however model can be used to calculate other microbe kinetics Can be used for process, design, optimization and scale-up (industrial purpose) to enhance ethanol productivity

20. Critique Take into account Cellulase inactivity Understand mechanisms of inhibition (i.e. feedback inhibition) Use larger concentrations of substrate (max = 120g/L)

21. References http://www.ncbi.nlm.nih.gov/pubmed/18609 632 http://www.ncbi.nlm.nih.gov/pubmed/18609 632 http://bio-process.com/wp- content/uploads/2009/12/Cellulosic-SSF.pdf