Tutorial 10
Design and analysis of continuous bioreactor
Date: 03/11/2025
1. A continuous stirred-tank bioreactor used for the production of bioethanol using
Baker’s yeast is continuously supplied with a total working volume of 2 L has a feed
flow rate of 0.5 liter per hour. At steady state, the biomass concentration in the reactor
is 1 g/L and the yeast follows the classic Monod growth kinetics with a maximum
specific growth rate of 0.4 h-1 and the Monod constant of 0.05 g/L. Assume the system
is operating with ideal mixing and steady state.
a. Calculate the dilution rate (D) and average residence time (τ) of the fermentation
broth.
b. Determine the special growth rate (μ) of the yeast in the chemostat.
c. Using the Monod equation, estimate the steady-state substrate concentration (S)
inside the reactor.
Solution
a. Calculate the dilution rate (D) and average residence time (τ) of the fermentation broth.
𝑭𝒍𝒐𝒘 𝒓𝒂𝒕𝒆(𝑭)
𝑫𝒊𝒍𝒖𝒕𝒊𝒐𝒏(𝑫) =
𝑽𝒐𝒍𝒖𝒎𝒆(𝑽)
0.5 L⁄h
Dilution(D) = = 0.25 h−1
2L
𝟏
𝑨𝒗𝒆𝒓𝒂𝒈𝒆 𝒓𝒆𝒔𝒊𝒅𝒆𝒏𝒄𝒆 𝒕𝒊𝒎𝒆(𝝉) =
𝑫𝒊𝒍𝒖𝒕𝒊𝒐𝒏(𝑫)
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Average residence time(τ) = =4h
0.25h−1
b. Determine the special growth rate (μ) of the yeast in the chemostat.
For chemostat working under steady state; μ = D = 0.25 h−1
c. Using the Monod equation, estimate the steady-state substrate concentration (S) inside the
reactor.
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𝝁𝑲𝒔
𝑺=
𝝁𝒎𝒂𝒙 − 𝝁
(0.25 h−1 ) × (0.05 g⁄L)
S=
(0.4 h−1 ) − (0.25 h−1 )
S = 0.0833 g⁄L
2. A continuous fermentation is carried out using Zymomonas mobilis in a chemostat
culture of 45 m3 fermenter and the feed contains 9.5 g/L of glucose. The organism has
a Monod constant of 0.35 g/L. The biomass yield coefficient (Y X/S) and product yield
coefficient (YP/S) are 0.09 g/g and 5.3 g/g respectively. The maximum specific growth
rate was observed to be 0.25 h-1 and Monod constant is 0.2 g/L .The value of
maintenance coefficient is 1.9 g g-1 h-1 and the value of Si is 9.5 g/L. The specific
product formation rate is 2.63 h-1.
a. What is the flow rate required to achieve a steady state substrate concentration of
1.2 g/L?
b. At the flow rate (a), what will be cell density?
c. At the same flow rate of (a), what is the concentration of the product produced?
d. Calculate the critical dilution (Dcrit).
Solution
a. What is the flow rate required to achieve a steady state substrate concentration of 1.2
g/L?
𝝁𝒎𝒂𝒙 𝑺
𝑫=𝝁=
𝑲𝒔 + 𝑺
(0.25h−1 )(1.2 g⁄L)
D= = 0.2143 h−1
(0.2 g⁄L) + (1.2 g⁄L)
𝑭𝒍𝒐𝒘 𝒓𝒂𝒕𝒆(𝑭) = 𝑫𝒊𝒍𝒖𝒕𝒊𝒐𝒏(𝑫) × 𝑽𝒐𝒍𝒖𝒎𝒆(𝑽)
Flow rate(F) = 0.2143 h × 45m3
Flow rate(F) = 0.2143 h−1 × 45m3
Flow rate(F) = 9.64 m3 ⁄h
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b. At the flow rate (a), what will be cell density?
𝑫(𝑺𝒊 − 𝑺)
𝒙=
𝑫
𝒀𝑿⁄𝑺 + 𝒎𝒔
(0.2143 h−1 )(9.5 g⁄L − 1.2 g⁄L)
x=
0.2143 h−1
+ 1.9 g g −1 h−1
0.09 g⁄g
x = 0.4158 g⁄L
c. At the same flow rate of (a), what is the concentration of the product produced?
𝒒𝒑 𝒙
𝒑 = 𝒑𝒊 + The value of pi is zero as the
𝑫
product concentration at the
(2.63 h−1 )(0.4158 g⁄L) initiation of the process is zero.
p=
0.2143 h−1
p = 5.10 g⁄L
d. Calculate the critical dilution (Dcrit).
𝝁𝒎𝒂𝒙 𝑺𝒊
𝑫𝒄𝒓𝒊𝒕 =
𝑲𝒔 + 𝑺𝒊
(0.25 h−1 )(9.5 g⁄L)
Dcrit =
(0.2 g⁄L) + (9.5 g⁄L)
Dcrit = 0.245 h−1
3. Tetrahymena thermophila protozoa have a minimum doubling time of 6.5 hours when
grown using bacteria as the limiting substrate. The yield of protozoa biomass is 0.33
g per gram of bacteria and the substrate constant is 12 mg/L. The protozoa are
cultured at steady state in a chemostat using a feed stream containing 10 g/L of
nonviable bacteria.
a. What is the maximum dilution rate for operation of the chemostat?
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b. What is the concentration of T. thermophila when the operating dilution rate is
one-half of the maximum?
c. What is the concentration of bacteria when the dilution rate is three-quarters of
the maximum?
d. What is the biomass productivity when the dilution is one-third of the maximum?
Solution
a. What is the maximum dilution rate for operation of the chemostat?
𝒍𝒏(𝟐) 0.693
𝝁𝒎𝒂𝒙 = = = 0.107 h−1
𝒕𝒅 6.5 h
𝝁𝒎𝒂𝒙 𝑺𝒊
𝑫𝒄𝒓𝒊𝒕 =
𝑲𝒔 + 𝑺𝒊
(0.107 h−1 )(10 g⁄L)
Dcrit =
(12 × 10−3 g⁄L) + (10 g⁄L)
Dcrit = 0.107 h−1
b. What is the concentration of T. thermophila when the operating dilution rate is one-half
of the maximum?
𝝁𝒎𝒂𝒙 0.107 h−1
𝑫= = = 0.535 h−1
𝟐 2
𝑫𝑲𝑺
𝒙 = (𝒔𝒊 − )𝒀
𝝁𝒎𝒂𝒙 − 𝑫 𝑿⁄𝑺
(0.535 h−1 )(12 × 10−3 g⁄L)
x = (10 g⁄L − ) 0.33 g⁄g
(0.107 h−1 ) − (0.535 h−1 )
x = 3.3 g⁄L
c. What is the concentration of bacteria when the dilution rate is three-quarters of the
maximum?
𝟑
𝑫= × 𝑫𝒄𝒓𝒊𝒕
𝟒
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D= × 0.107 h−1 = 0.08 h−1
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𝑫𝑲𝒔
𝑺=
𝝁𝒎𝒂𝒙 − 𝑫
(0.08 h−1 )(12 × 10−3 g⁄L)
S=
(0.107 h−1 ) − (0.08 h−1 )
S = 0.036 g⁄L
d. What is the biomass productivity when the dilution is one-third of the maximum?
𝑫𝒄𝒓𝒊𝒕 0.107 h−1
𝑫= = = 0.036 h−1
𝟑 3
𝑫𝑲𝒔
𝑸𝑿 = 𝑫 (𝒔𝒊 − )𝒀
𝝁𝒎𝒂𝒙 − 𝑫 𝑿⁄𝑺
(0.036 h−1 )(12 × 10−3 g⁄L)
Q X = (0.036 h−1 ) (10 g⁄L − ) 0.33 g⁄g
(0.107 h−1 ) − (0.036 h−1 )
Q X = 0.119 g L−1 h−1
4. A bioprocess engineer operates a chemostat with cell recycle to maximize biomass
productivity. The following parameters are observed;
Maximum specific growth rate – 0.5 h-1
Saturation constant – 0.2 g/L
Yield coefficient – 0.5 g/g
Inlet substrate concentration – 20 g/L
Recycle ratio – 0.5
Biomass concentration factor – 2
Dilution rate – 0.4 h-1
Using the above-mentioned parameters, calculate the following;
a. Steady state substrate concentration in the reactor
b. Steady state biomass concentration with recycle
c. Volumetric biomass productivity for the chemostat with cell recycle
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Solution
a. Steady state substrate concentration in the reactor
𝑫𝑲[𝟏 + 𝜶(𝟏 − 𝜷)]
𝑺=
𝝁𝒎𝒂𝒙 − 𝑫[𝟏 + 𝜶(𝟏 − 𝜷)]
(0.4 h−1 )(0.2 g⁄L)[1 + 0.5(1 − 2)]
S=
(0.5 h−1 ) − (0.4 h−1 )[1 + 0.5(1 − 2)]
0.04 g⁄L
S=
0.3
S = 0.134 g⁄L
b. Steady state biomass concentration with recycle
𝒀𝑿⁄𝑺 𝑫𝑲[𝟏 + 𝜶(𝟏 − 𝜷)]
𝑿𝟏 = [𝒔𝒊 − ]
[𝟏 + 𝜶(𝟏 − 𝜷)] 𝝁𝒎𝒂𝒙 − 𝑫[𝟏 + 𝜶(𝟏 − 𝜷)]
0.5 𝑔⁄𝑔
X1 = [(20 g⁄L) − (0.134 g⁄L)]
0.5
X1 = 19.87 g⁄L
c. Volumetric biomass productivity for the chemostat with cell recycle
𝑸𝑿 = 𝑫𝑿𝟏
𝒀𝑿⁄𝑺 𝑫𝑲[𝟏 + 𝜶(𝟏 − 𝜷)]
𝑸𝑿 = 𝑫. [𝒔𝒊 − ]
[𝟏 + 𝜶(𝟏 − 𝜷)] 𝝁𝒎𝒂𝒙 − 𝑫[𝟏 + 𝜶(𝟏 − 𝜷)]
Q X = (0.4 h−1 )(19.87 g⁄L)
Q X = 7.95 g L−1 h−1
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