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Internal Use: Bio-Immobilization and Mass Transfer Effects

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Internal Use: Bio-Immobilization and Mass Transfer Effects

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businesskylamiel
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© © All Rights Reserved
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Internal Use

Worksheet 3 (100 points)


CH148 BIO-IMMOBILIZATION AND MASS TRANSFER EFFECTS
BIOCHEMICAL
ENGINEERING Group No.: # Rating: 100/100

General Instructions
1. Fill in the details required in this activity form and attach it before the solution sheet as a PDF file.
2. Use the file name CourseCode-Section-Term-GroupNumber-ActivityCode (Example: CH100-A1-1T2425-G1-WS1).
3. Attempt to answer individually and discuss with your groupmates afterward.
4. Academic Dishonesty in any form is prohibited in the entirety of this course. All infarctions within the Mapúa University policies on
Academic Integrity, AI Use, and other related shall be faced with the gravest consequences.

Group Members (complete name: Ln, Fn M.I.)

A. Problem Solving. Solve the problems clearly, completely, and correctly. Incomplete (or shortcut) solutions will
not be given full credit regardless of the final answers. Alternative methods in this section are permitted. Box
your final answer(s).

1. (50 points) L-Lactate 2-monooxygenase from Mycobacterium smegmatis is immobilized in spherical agarose beads.
The enzyme catalyzes the reaction: C 3 H 6 O 3 +O 2 → C2 H 4 O 2 + H 2 O+C O 2. Agarose beads 4 mm in diameter are
immersed in a well-mixed solution containing 0.5 mM oxygen. A high lactic acid concentration is provided so that
oxygen is the rate limiting substrate. The effective diffusivity of oxygen in agarose is 2.1 × 10 -9 m2 s-1. Km for the
immobilized enzyme is 0.015 mM and vm is 0.12 mol s-1 per kg of enzyme. The beads contain 0.012 kg of enzyme m -3 of
gel. External mass transfer effects are negligible.
a. Plot the oxygen concentration profile inside the beads.
b. What fraction of the catalyst volume is active?
c. Determine the largest bead size that allows the maximum overall rate of conversion.

2. (50 points) Invertase, which catalyzes the reaction sucrose+ water → glucose+fructose , from Aspergillus oryzae
is immobilized in porous resin particles of diameter 1.6 mm at a density of 0.1 μmol of enzyme g -1. The effective
diffusivity of sucrose in the resin is 1.3 × 10-11 m2 s-1. The resin is placed in a spinning basket reactor operated so that
external mass transfer effects are eliminated. At a sucrose concentration of 0.85 kg m -3, the observed rate of
conversion is 1.25 × 10-3 kg s-1 m-3 of resin. Km for the immobilized enzyme is 3.5 kg m-3.
a. Calculate the effectiveness factor.
b. Determine the true first-order reaction constant for immobilized invertase.
c. Assume that the specific enzyme activity is not affected by steric hindrance or conformational changes as the
enzyme loading increases. This means that k 1 should be directly proportional to the enzyme concentration in the
resin. Plot the changes in effectiveness factor and observed reaction rate as a function of enzyme loading from 0.01
μmol g-1 to 2.0 μmol g-1. Comment on the relative benefit of increasing the concentration of enzyme in the resin.
Internal Use
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