TUTORIAL

Chapter 5

QUESTION 1

Urease with molecular weight of 482 700, belong to the superfamily of amidohydrolases and
phosphotriesterases. It is an enzyme that catalyzes the hydrolysis of urea into carbon dioxide and
ammonia. The urease solution was produced through the active site with the protein binding
(MW = 726 300) containing 21 g protein/200 mL solution and form a tetrahedral cluster that fills
the cavity site through hydrogen bonds. The viscosity of urease solution is outlined in Table 1.

Table 1: Viscosity of urease solution.

Temperature Viscosity (centipoises)
(ºC)
20 1500
40 1010
60 520
80 30

a) Determine the temperature (in oC) of the urease solution so that the diffusivity of urease
diffusing in 1255 centipoises solution is 4.96 x 10-11 m2/s. (VA = 1.14x 10-7 m3/kg mol)
(8 Marks)

b) Assuming no binding to the protein solution, calculate the flux of urease at 72 oC if the
original amount of urease in the solution along 0.25 cm distance is 0.5 g mol/mL. State
the assumption to support your calculation.
(18 Marks)

c) Repeat question b) in (kg mol/cm2.hr), however 30% of the urease is required to bind to
the protein in the solution.
(14 Marks)
QUESTION 2

Glycerol with formula structure of C3H8O3 (Figure 1) is also known as glycerin diffusing in
water solution at 37oC to develop tryptone glucose yeast (TGY) in laboratory. In order to
produce TGY with good performance, 6.02 wt% of agar gel was originally developed at 37oC as
a general medium for glycerol-water solution. The viscosity of glycerol-agar solution is outlined
in Table 2. If the original amount of glycerol in the agar is 0.3 g mol per liter solution, estimate:

Figure 1: Molecular structure of glycerol (glycerin).

Table 2: Viscosity of glycerol-agar solution.

Temperature (ºC) Viscosity (Pa.s)
5 2.8
20 9.21
40 20.96

a) The diffusivity of the glycerol in the water solution.

(10 Marks)
b) The diffusivity of the glycerol in the 6.02 wt % of agar at 37oC by using the temperature-
corrected factor from the Stokes-Einstein equation.
(12 Marks)
c) Then, the researchers would like to produce TGY from the glycerol-water with protein
molecule (MW = 63 000) instead of agar gel medium where 25% of the glycerol bind to
the protein. Based on the value of glycerol-water diffusivity in question a), calculate the
flux of glycerol if the amount of glycerol in the protein solution is 0.3 g mol per liter and
the distance is approximately 0.5 cm. State the assumption to support your calculation.
(cp = 12 kg/m3)
(18 Marks)
QUESTION 3

Sucrose with the molecular formula C12H22O11 is a common saccharide found in many plants and
often extracted from sugarcane for human consumption. When sucrose is consumed, the enzyme
beta-fructosidase separates sucrose into its individual sugar units of glucose and fructose.
However, it is necessary to dissolve the sucrose in an aqueous solution containing 21 g
protein/200mL solutionat 25oCto control the undesirable of enzyme beta-fructosidase. The
viscosity of aqueous solution at different temperature is presented in Table 3.

Table 3: Viscosity of aqeous solution

Temperature (ºC) Viscosity, µ×103
(kg/m.s)
24 0.981
37 0.680
48 0.382

a) By using temperature corrected factor, determine the diffusion coefficient for sucrose
solute in aqueous solution at 25oC.
(8 Marks)

b) If the aqueous solution containing the bovine serum albumin (BSA) as a protein,
calculate the diffusivity of BSA in aqueous solution at 25oC. Calculate the error of the
estimate calculation with the literature value.
(8 Marks)

c) Based on the value of diffusivity in question a) and b), calculate the flux of sucrose in the
protein solution if 75% of the solutes not bound to the protein. The amount of sucrose in
the protein solution at point (1) and point (2) are 300 g mol/m3 and 15 g mol/m3,
respectively, and the distance is approximately 0.5 cm.
(14 Marks)