1. Enzyme is immobilized in 8 mm diameter agarose beads at a concentration of 0.

018 kg protein m 3-
gel. Ten beads are immersed in a well-mixed solution containing 3.2x10-3 kg m-3 substrate. The
effective diffusivity of substrate in agarose gel is 2.1x10 -9 m2 s-1. Kinetics of the enzyme can be
approximated as first order with specific rate constant 3.11x105 s-1 per kg protein. Mass transfer
effects outside the particles are negligible. Plot the steady-state substrate concentration profile as a
function of particle radius.

[GIVEN]
 Bead diameter = 8 mm
 Concentration of enzyme in gel = 0.018 kg protein m3-gel
 Number of beads immersed in well-mixed solution = 10
 Concentration of substrate in the solution = 3.2x10-3 kg m-3 substrate
 Effective Diffusivity (DAe) of substrate in agarose gel = 2.1x10-9 m2 s-1
 Specific rate constant (k) =3.11x105 s-1 per kg protein
[REQUIRED]
Steady-state substrate concentration profile as a function of particle radius
[ASSUMPTION]
 Mass transfer effects outside the particles are negligible.
 The beads are spherical.
 The effective diffusivity (DAe) is constant and independent of substrate concentration in the
particle.
 The particle is isothermal such that the kinetic parameters are constant.
[SOLUTION]
Doing a steady-state shell mass balance,
{𝑚𝑎𝑠𝑠 𝑎𝑐𝑐𝑢𝑚𝑢𝑙𝑎𝑡𝑒𝑑 𝑤𝑖𝑡ℎ𝑖𝑛 𝑡ℎ𝑒 𝑠𝑦𝑠𝑡𝑒𝑚} = {𝑚𝑎𝑠𝑠 𝑖𝑛 𝑡ℎ𝑟𝑜𝑢𝑔ℎ 𝑡ℎ𝑒 𝑠𝑦𝑠𝑡𝑒𝑚} −
{𝑚𝑎𝑠𝑠 𝑜𝑢𝑡 𝑡ℎ𝑟𝑜𝑢𝑔ℎ 𝑡ℎ𝑒 𝑠𝑦𝑠𝑡𝑒𝑚} + {𝑚𝑎𝑠𝑠 𝑔𝑒𝑛𝑒𝑟𝑎𝑡𝑒𝑑 𝑤𝑖𝑡ℎ𝑖𝑛 𝑡ℎ𝑒 𝑠𝑦𝑠𝑡𝑒𝑚} −
{𝑚𝑎𝑠𝑠 𝑐𝑜𝑛𝑠𝑢𝑚𝑒𝑑 𝑤𝑖𝑡ℎ𝑖𝑛 𝑡ℎ𝑒 𝑠𝑦𝑠𝑡𝑒𝑚} (1)
Where:
𝑑𝐶𝐴
 Rate of input by diffusion: (𝐷𝐴𝑒 4𝜋𝑟 2 )|
𝑑𝑟 𝑟+∆𝑟
𝑑𝐶𝐴
 Rate of output by diffusion : (𝐷𝐴𝑒 2
4𝜋𝑟 )|
𝑑𝑟 𝑟
 Rate of generation: 0
 Rate of consumption by reaction : 2
𝑟𝐴 4𝜋𝑟 ∆𝑟
 Rate of accumulation at steady state: 0
Substituting,
𝑑𝐶𝐴 𝑑𝐶𝐴
(𝐷𝐴𝑒 4𝜋𝑟 2 )| − (𝐷𝐴𝑒 4𝜋𝑟 2 )| − 𝑟𝐴 4𝜋𝑟 2 ∆𝑟 = 0 (2)
𝑑𝑟 𝑟+∆𝑟 𝑑𝑟 𝑟
𝑑𝐶𝐴 2 𝑑𝐶
(𝐷𝐴𝑒 𝑟 )| −(𝐷𝐴𝑒 𝐴𝑟 2 )|
𝑑𝑟 𝑑𝑟
𝑟+∆𝑟 𝑟
− 𝑟𝐴 𝑟 2 = 0 (3)
∆𝑟
𝑑𝐶𝐴 2
∆(𝐷𝐴𝑒 𝑟 )
𝑑𝑟
− 𝑟𝐴 𝑟 2 = 0 (4)
∆𝑟

Taking the limit of equation (4) as ∆𝑟 → 0,

𝑑 𝑑𝐶𝐴
(𝐷𝐴𝑒 𝑟 2 ) − 𝑟𝐴 𝑟 2 = 0 (5)
𝑑𝑟 𝑑𝑟

Since 𝐷𝐴𝑒 is assumed to be constant and independent of substrate concentration in the particle, it
can be moved outside the differential,
𝑑 𝑑𝐶
𝐷𝐴𝑒 𝑑𝑟 ( 𝑑𝑟𝐴 𝑟 2 ) − 𝑟𝐴 𝑟 2 = 0 (6)
 Rewriting Equation (6) in its expanded form,
𝑑2 𝐶𝐴 𝑑𝐶𝐴
𝐷𝐴𝑒 ( 𝑟 2 + 2𝑟 ) − 𝑟𝐴 𝑟 2 = 0 (7)
𝑑𝑟 𝑑𝑟

For first order reactions, equation (7) becomes

𝑑2 𝐶𝐴 𝑑𝐶𝐴
𝐷𝐴𝑒 ( 𝑟 2 + 2𝑟 ) − 𝑘1 𝑟 2 = 0 (8)
𝑑𝑟 𝑑𝑟

Since equation (8) is a second order differential equation, two boundary conditions are needed.
These conditions are:
𝐶𝐴 = 𝐶𝐴,𝑆 at r=R
𝑑𝐶𝐴
=0 at r=0
𝑑𝑟

where 𝐶𝐴,𝑆 is the concentration of the substrate at the outer surface of the particle.
Integrating equation (8) with the two boundary conditions gives the following equation:
𝑘
sinh(𝑟√ 1 )
𝑅 𝐷𝐴𝑒
𝐶𝐴 = 𝐶𝐴,𝑆 𝑟 𝑘
(9)
sinh(𝑅√ 1 )
𝐷𝐴𝑒

Using equation (9) to obtain the plot of the substrate concentration as a function of r at steady state,

𝑘1
sinh(𝑟√
𝑘𝑔 (4 −3
× 10 𝑚) 𝐷𝐴𝑒 )
𝐶𝐴 = (3.2 × 10−3 )
𝑚3 𝑟 𝑘
sinh(𝑅√𝐷 1 )
𝐴𝑒

Determining k,
𝑘 = 𝑘 ′ (𝑎𝑚𝑜𝑢𝑛𝑡 𝑜𝑓 𝑒𝑛𝑧𝑦𝑚𝑒 𝑖𝑛 𝑘𝑔)
4 𝑘𝑔
𝑘 = (2.68 × 10−6 𝑚3 ) ( 𝜋(4 × 10−3 𝑚)3 ) (0.018 3 )
3 𝑚
𝑘 = 0.015 𝑠 −1
Evaluating the denominator term:
Recall:
𝑒 𝑥 − 𝑒 −𝑥
sinh 𝑥 =
2
𝑘
Where: 𝑅√𝐷 1 = 10. 693
𝐴𝑒

𝑘1 𝑒 10.693 − 𝑒 −10.693
sinh 𝑅√ =
𝐷𝐴𝑒 2

𝑘1
sinh 𝑅√ = 2.202 × 104
𝐷𝐴𝑒

Going back,
0.015
𝑘𝑔 (4×10−3 𝑚) sinh(𝑟√2.1×10−9 )
𝐶𝐴 = (3.2 × 10−3 𝑚3 ) (10)
𝑟 2.202×104

Equation (10) is then used to plot 𝐶𝐴 at r=0 until r=R=4 mm. The resulting plot is shown below.
 1.20E-06

Substrate Concentration, CA (kg/m3) 1.00E-06

8.00E-07

6.00E-07

4.00E-07

2.00E-07

0.00E+00
0.0000 0.0010 0.0020 0.0030 0.0040
radius, r (m)

Figure 1. Substrate concentration (CA) as a function of particle radius, r

From Figure 1, the substrate concentration drops rapidly inside the particle as shown by the
decreasing curve from r=4 mm.