ChE513: Biochemical Engineering University of Santo Tomas Engr.

Achilles Espaldon

Submission Date: June 13, 2012

Assignment # 1 (for all sections) 1. A modified reaction velocity or rate equation below instead of the Michaelis-Menten (M-M) model is used to predict conversion in a batch reactor. The initial substrate concentration is 10-5uM, km = 2x10-3 uM and vm = 6.77x10-7 uM/s with value of ks = 0.1uM as constant; vm .S v S 2 ' k m S KS a. Derive the new equation for calculating the time for reaction. b. How long (in min.) shall I run the reaction to obtain an 80% conversion? c. Which substrate concentration in this given set gives the highest reaction velocity? (S in uM = 0.08, 0.02, 0.014, 0.0044, 5x10-4) d. Plot the v vs. [S] graph. Comment on the result.
2. 3. Derive an equation to calculate the volume of a reactor in a batch process using M-M model.

An enzyme with km = 2.6x10-3M was assayed at an initial substrate concentration of 0.3M. The observed velocity was 5.9x10-5M/min. If the initial concentration of substrate is 2x10-5M, a. what would be the product concentration after 90 min.? b. determine the yield for 90 min. c. construct a plot of the concentration of substrate and product with time for 5 data points (1, 10, 30, 50, 70, 90 min.) d. to obtain a 98% conversion, how long shall I wait for my reaction to achieve such conversion? e. comment on the order of the reaction
An enzyme ATPase (MW = 5x104 Da) with km = 10-4M and k2 = 104/min at 370C. the reaction is catalyzed as;
ATPase ATP ADP P

4.

Which is similar to

k1 , E S k 1 E.S k 2 P E

Where S is the ATP. The enzyme at this temperature is unstable. The enzyme inactivation kinetics are first order described as;

E Eo .e k d .t
Where Eo is the initial enzyme concentration and kd = 0.1/ min. In an experiment with a partially pure enzyme preparation, 10ug of total crude protein (containing enzyme) is added to a 1 mL reaction mixture containing 0.02M ATP and incubated at 370C. After 12 hours the reaction ends at t = infinity and the inorganic phosphate P concentration is found to be 0.002M, which was initially zero. What fraction of the crude protein preparation was the enzyme? (hint: since [S] >>> km the reaction rate can be represented by;

v
5.

dP k2 .[E ] k2 .[Eo ].e k d .t dt

Answer # 6 in the file Enzyme kinetics examples and problems.

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