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
SSF Kinetics Cellulose cellobiose glucose r1r1 r2r2
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
How do we measure enzyme activity as an uncoupled system? 3g/L
We can simplify the volumetric rate constants = constant
Effect of cellobiose on cellulase activity
Avg. Effect of cellobiose on B-glucosidase K 2B ?
Effect of cellobiose on B-glucosidase Maximal rate
Glucose concentration and enzyme activity
Ethanol concentration and Enzyme activity K 2E = NEGLIGIBLE
Summary
Putting it all together
Putting it all together continued Almost perfect fit but not yet Why?
First order rate loss added to model Fail
(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 content/uploads/2009/12/Cellulosic-SSF.pdf