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Enzymatic Hydrolysis of cellulose for production of fuel ethanol by SSF. Presented by Alaaedine Talab. What is SSF?. The reaction. The reaction continued. β(1→ 4). Purpose of SSF. Minimize Hydrolysis byproducts Economically favorable Minimize inhibitory factors Eliminate transfer time.
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Enzymatic Hydrolysis of cellulose for production of fuel ethanol by SSF Presented by AlaaedineTalab
The reaction continued β(1→ 4)
Purpose of SSF • Minimize Hydrolysis byproducts • Economically favorable • Minimize inhibitory factors • Eliminate transfer time
SSF Kinetics r1 r2 glucose cellobiose Cellulose
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 • TrichodermaReesei • Initial rates determined after enzyme deactivation(20 minutes boiling water, denatur) 15s intervals 1 minute duration
How do we measure enzyme activity as an uncoupled system? 3g/L
Effect of cellobiose on B-glucosidase Avg. K2B?
Effect of cellobiose on B-glucosidase Maximal rate
Ethanol concentration and Enzyme activity K2E = NEGLIGIBLE
Putting it all together continued • Almost perfect fit but not yet • Why?
(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
Critique • Take into account Cellulase inactivity • Understand mechanisms of inhibition (i.e. feedback inhibition) • Use larger concentrations of substrate (max = 120g/L)
References • http://www.ncbi.nlm.nih.gov/pubmed/18609632 • http://bio-process.com/wp-content/uploads/2009/12/Cellulosic-SSF.pdf