04EnzymesParameterEstimation - Engineers Scientists BME...

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1 Engineers – Scientists BME – Bridges between Scientist …. What is the mechanism of catalysis? What are hypothesis I can think of? What are clever experiments to test this? Engineers …. How can I maximize the use of this enzyme? What factors affect reactions (heat, mixing, ..) How can I quantitatively model and optimize? 2 Enzymes Structure and Mechanism of Catalysis S P S G G o
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3 Single Substrate Michaelis- Menten Kinetics S ubstrate P roduct E nzyme [E] : Brackets means concentration of that thing 4 1. Rate of product accumulation increases over time. (first phase greatly exaggerated; usually can’t see) 2. Product concentration increases linearly with time. 3. Substrate is depleted, so the curve starts to level off. How does product concentration change with time? Constant [E]; [S] at time 0 = S 0 ; [P] at time 0 = 0 V
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How does enzyme velocity change with [S]? Constant [E]; Velocity V = dP/dt Michaelis-Menten Equation: Vmax: Limiting velocity at high [S] Km: [S] that leads to half-maximal velocity m S K S v v + = max , 6 Michaelis-Menten Kinetics Henri (1902): Theoretical explanation (Henri, Michaelis, Menten 1913) o m E v S K S v v α = + = max max , [ ][ ] [ ] [ ] o m o m k k k E k v S K S v ES k v ES E E K k k ES E S E P ES E S 2 max max 2 1 1 , , 2 1 1 = + = = + = = = + → + Assume equilibrium: Dissociation constant
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