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PROBLEMS WITH SOLUTIONS
IN BIOCHEMICAL ENGINEERING
Compiled by:
Abuan, Rebecca Masaoay, Josafet
Acod, Jullienne Meria, Shirvin
Amparo, Jude Nonog, Charisse
Agudo, Anna Leslie Notarte, Melanie
Curaming, Felyn Pineda, Angelica Ara
Dela Cruz, Henry Rondina, Ryan
Duclayan, Micadel Talavera, Yul Sses
Ebil, Ephraim Tapiador, Jonayka
Espartinas, Christian Gabriel Valencia, Laurice Kay
Garoy, Angelica Valencia, Pamela
Gonzales, Katrina Mae Vedaña, Marvin
Javar, Maria Camille Villegas, CJ
Labasan, John Paul
Presented to:
Engr. Janice Omadto
December 201
CHAPTER 2: ENZYME KINETICS
2.3 In some enzyme- catalyzed reaction, multiple complexes are involved as follows:
S + E ↔ ( ES )1
( ES )1 ↔ ( ES )2
( E )2 → P + E
REQUIRED: Develop a rate expression using
a. Michaelis Menten approach
b. The Briggs Haldane approach
SOLUTION :
−dcs dcp
rp = = = ksCes − k4CpCe
dt dt
Ceo = Ce = Ces ; Ce = Ceo – Ces
rp = k3Ces – k4CpCeo + k4CpCes
rp = ( k3 + k4 ) Ces – k4CpCeo
k1 CsCe = k2Ces
k1(Cs)(Ceo-Ces) = k2Ces
k1CsCeo-k1CsCes=k2Ces
k2Ces + k1CsCes = k1CsCeo
k1 CsCeo
Ces =
k2 + k1Cs
CeoCs
Ces =
k2
+ Cs
k1
CeoCs
rp = ( k3 + k4Cp) k2 - k4Ceo
+Cs
k1
k2
(ks+k4Cp)(CeoCs)− ( Cs ) ( k4CpCeo )
k1
rp = k2
+Cs
k1
k2k4CpCeo
k3CeoCs + k4CeoCpCs − − k4CpCesCs
rp = k1
k2
+ Cs
k1
k2k4CpCeo
k3CeoCs −
rp = k1
k2
+ Cs
k1
k2k4
Ceo (k3cs − Cp)
rp = k1
k2
+ Cs
k1
Ce=Ceo=Ces
K1Cs ( Ceo-Ces) = k2Ces
K2Ces + k1CsCes = k1CsCeo
k1CsCeo
k2
+ Cs
k1
CsCeo
k2
+ Cs
k1
ANSWER
k3CeoCs rmaxCx
=
k2
+ Cs Km + Cs
k1
The Briggs Haldane
−dcs dcp
= = k3Ces
dt dt
Rate determining equation:

−dcs
= k1CsCe − k2Ces − k3Ces = 0
dt
Ceo = Ce + Ces
K1Cs ( Ceo-Ces )=k2Ces-k3Ces
K1CsCeo-k1CsCes-k3 ; Ces=0
K1CsCes+k2Ces+k3Ces = k1CsCeo
k1CsCeo
Ces =
k1Cs + k2 + k3
CsCeo
Ces =
k3 + k2
+ Cs
k1
−dcs dcp k3CsCeo

= =
dt dt k3 + k2
+ Cs
k1
rmaxCs
rp= KmCs
I
2.4 Eadie (1942) measured the initial reaction rate of hydrolysis of acetylcholine (substrate) by
dog serum (source of enzyme) and obtained the following data:
Substrate Concentration Initial Reaction Rate

mol/L mol/L·min
0.0032 0.111
0.0049 0.148
0.0062 0.143
0.0080 0.166
0.0095 0.200
Evaluate the Michaelis-Menten kinetic parameters by employing (a) the Langmuir plot, (b) the
Lineweaver-Burk plot, and (c) the Eadie-Hofstee plot.
Given: *refer to table

Required: Michaelis-Menten kinetic parameters
Solution:
a. Langmuir
CS KM 1
= + CS
r rmax rmax
x, CS y, CS/r
0.0032 0.0032/0.111
0.0049 0.0049/0.148
0.0062 0.0062/0.143
0.0080 0.0080/0.166 rmax = 0.3018 mol/L·min
0.0095 0.0095/0.200 KM = 5.7721 x 10-3 mol/L
b. Lineweaver-Burk
1 1 KM 1
= +
r rmax rmax CS
x, 1/CS y, 1/r
1/0.0032 1/0.111
1/0.0049 1/0.148
1/0.0062 1/0.143
1/0.0080 1/0.166 rmax = 0.2752 mol/L·min
1/0.0095 1/0.200 KM = 4.7303 x 10-3 mol/L
c. Eadie-Hofstee
r
r = rmax - KM
CS
x, r/CS y, r
0.111/0.0032 0.111
0.148/0.0049 0.148
0.143/0.0062 0.143
0.166/0.0080 0.166 rmax = 0.2645 mol/L·min
0.200/0.0095 0.200 KM = 4.2731 x 10-3 mol/L
2.7 The KM value of an enzyme is known to be 0.01 mol/L. To measure the maximum reaction rate
catalyzed by the enzyme, you measured the initial rate of the reaction and found that 10 percent of
the initial substrate was consumed in 5 minutes. The initial substrate concentration is 3.4x10-4
mol/L. Assume that the reaction can be expressed by the Michaelis-Menten kinetics.
a. What is the maximum reaction rate?
b. What is the concentration of the substrate after 15 minutes?
Given:
KM = 0.01 mol/L @t=5 minutes
Cso= 3.4x10-4 mol/L 10 percent was consumed
Required: a.) rmax

b.) Cs @t=15 minutes
Solution:
K M ln 𝐶𝑠𝑜
𝐶𝑠
+ (𝐶𝑠𝑜−𝐶𝑠) = 𝑟𝑚𝑎𝑥 𝑡
mol
0.01 1
ln 0.9 + (1−0.9)(3.4x10−4)
𝑚𝑜𝑙
𝐿
= 𝑟𝑚𝑎𝑥 (5𝑥60)𝑠
L
𝑟𝑚𝑎𝑥 = 3.6254x10-6 kmol/m3 s

@t= 15 minutes
K M ln 𝐶𝑠𝑜
𝐶𝑠
+ (𝐶𝑠𝑜−𝐶𝑠) = 𝑟𝑚𝑎𝑥 𝑡
mol kmol
0.01 ln 3.4𝑥10−4
𝐶𝑠
+ (3.4𝑥10−4−𝐶𝑠)
𝑚𝑜𝑙
𝐿
= (3.6254x10 − 6 m3 s ) (15𝑥60)𝑠
L
Cs= 2.4762 x10-4 mol/ L

2.8 A substrate is converted to a product by the catalytic action of an enzyme. Assume that the
Michaelis-Menten kinetic parameters for enzyme reaction are:
KM = 0.03 mol/L
rmax= 13 mol/ L min
a. What should be the size of a steady-state CSTR to convert 95 percent of incoming

substrate (Cso= 10 mol/L) with a flow rate of 10 L/h?
b. What should be the size of the reactor if you employ a plug-flow reactor instead of the
CSTR in part (a)?
Given: Req'd:
KM = 0.03 mol/L a)VCSTR
rmax= 13 mol/ L min b) VPFR
Cso= 10 mol/L
F= 10 L/h
Sol'n:
a)
𝐹 1 𝑟𝑚𝑎𝑥 𝐶𝑠
= =
𝑉 𝜏 (𝐶𝑠𝑜 − 𝐶𝑠)(𝐾𝑚 + 𝐶𝑠)
𝐹
𝑉=
𝑟𝑚𝑎𝑥 𝐶𝑠
(𝐶𝑠𝑜 − 𝐶𝑠)(𝐾𝑚 + 𝐶𝑠)
10𝐿 1ℎ𝑟
(
) (60min)
𝑉= ℎ
𝑚𝑜𝑙 𝑚𝑜𝑙
(13 𝐿 𝑚𝑖𝑛) (0.05 × 10 𝐿 )
10𝑚𝑜𝑙 0.5𝑚𝑜𝑙 0.03𝑚𝑜𝑙 0.5𝑚𝑜𝑙
( 𝐿 − 𝐿 )( + 𝐿 )
𝐿
VCSTR=0.1291 L
b)
𝐶𝑠𝑜 − 𝐶𝑠 𝑡
= −𝐾𝑚 + 𝑟𝑚𝑎𝑥 ( )
𝐶𝑠𝑜 𝐶𝑠
ln( 𝐶𝑠 ) ln (𝐶𝑠𝑜)
10 − 0.5 𝑡
= −(0.03) + (13) ( )
10 10
ln ( ) ln ( )
0.5 0.5
t = 0.7377 min
t=V/F
V=Ft
V=(0.7377 min)(1 h/ 60 min)(10 L/h)
VPFR = 0.1229 L
2.9 A substrate is decomposed in the presence of an enzyme according to the michaelis menten
equation with the following kinetic parameters:
grams
Km=10 liter
g
Rmax = 7 L−min
If we operate two 1-L CSTR n series at steady state, wht will be the concentration of substrate
leaving the second reactor? The flow rate is 0.5 L/min. The inlet substrate concentration is 50g/L
and the enzyme concentration in the two reactors is maintained in the sa value all of the time. Is
the two reactor system more efficient than one reactor whose volume is equal to the sum of the
two reactors?
GIVEN:
Cso
50g/L
Km= 10g/L
rmax= 7g/L-min
F= 0.5 L/min
REQUIRED:
a. Cs2
b. Is two reactor more efficient than 1 reactor with volume = 2L
rmax Csτ
Cs=-km + Cso−Cs
For the first reactor solve Cs1
7g 1l
( )(Cs2)( 0.5g )
L
min
Cs1= (-10g/L) + 50g
− Cs1
L
Cs1= 38.8650g/L
At the second reactor ; cso =38.8650g/L

7g 1l
( )(Cs2)( 0.5g )
L
min
Cs2= (-10g/L) + 38.8650− Cs2
Cs2= 28.50120g/L
Which is more efficient
Cso−Cs
% conversion = x 100%
Cso
50−28.5012
%conversion = x 100%
50
%conversion = 42.9976 % (for 2 CSTR IN SERIES)
For 1 CSTR with volume =2L

7g 2l
( )(Cs2)( 0.5g )
L
min
Cs= (-10g/L) + 50g
− Cs1
L
Cs= 29.1517 g/L
50 − 29.1517
%conversion = x 100%
50
Conversion = 41.6969 %% ( for 1 CSTR with 2L volume)

2.14 Eadie (1942) measured the initial reaction rate of hydrolysis of acetylcholine (substrate) by
dog serum (source of enzyme) in the absence and presence of prostigmine (inhibitor), 1.5 x 10-7
mol/L and obtained the following data:
Substrate Concentration Initial Reaction Rate Rate (mol/L.min)
(mol/L) Absence of Prostigmine Presence of Prostigmine
0.0032 0.111 0.059
0.0049 0.148 0.071
0.0062 0.143 0.091
0.0080 0.166 0.111
0.0095 0.200 0.125
a. Is prostigmine competitive or noncompetitive inhibitor?

b. Evaluate the Michaelis-Menten kinetic parameters in the presence of inhibitor by
employing the Langmuir plot.
0.09
0.08
y = 2.9883x + 0.0489
0.07
0.06
0.05 y = 3.3133x + 0.0191
Cs/r
0.04
0.03
0.02
0.01
0
0 0.001 0.002 0.003 0.004 0.005 0.006 0.007 0.008 0.009 0.01
Cs
Series1 Series2 Linear (Series1) Linear (Series2)
a. Prostigmine is a competitive inhibitor.

b. Rmax= 1/m = 0.3346 (with inhibitor concentration)
Rmax= 1/m = 0.3018 (no inhibitor concentration)
Km = bRmax = 5.7644x10-3
KI = bRmax = 0.0164
2.17 The initital rate of reaction for the enzymatic cleavage of deoxyguanosine triphosphate was
measured as a function of initial substrate concentration as follows (Kornberg et al., J. Biol. Chem.,
233, 159, 1958):
Given:
Substrate Concentration Initial Reaction Rate
μmol/L μmol/L min
6.7 0.30
3.5 0.25
1.7 0.16
a. Calculate the Michaelis-Menten constants of the above reaction.
b. When the inhibitor was added, the initial reaction rate was decreased as follows:
Substrate Inhibitor Initial Reaction Rate
μmol/L Μmol/L Μmol/L min
6.7 146 0.11
3.5 146 0.08
1.7 146 0.06
Is this competitive inhibition or noncompetitive inhibition? Justify your answer by showing the
effect of the inhibitor graphically. [Contributed by Professor Gary F. Bennett, The university of
Toledo, Toledo, OH]
Required: MM constants
Without Inhibitor With inhibitor

a) Langmuir a) Langmuir
𝐶𝑠 𝑘𝑚 1 𝐶𝑠 𝑘𝑚 1
= + (𝐶 ) = + (𝐶 )
𝑟 𝑟𝑚𝑎𝑥 𝑟𝑚𝑎𝑥 𝑠 𝑟 𝑟𝑚𝑎𝑥 𝑟𝑚𝑎𝑥 𝑠
r = 0.9968 μmol/L-min r = 0.9916 μmol/L-min

r(max) = 0.4215 μmol/L-min r(max) = 0.1567 μmol/L-min
km = 3.6317 μmol/L km = 2.9807 μmol/L
b) Lineweaver Burk b) Lineweaver Burk
1 1 𝑘𝑚 1 1 1 𝑘𝑚 1 1
= + ( ) = + ( )
𝑟 𝑟𝑚𝑎𝑥 𝑟𝑚𝑎𝑥 𝐶𝑠 𝑟 𝑟𝑚𝑎𝑥 𝑟𝑚𝑎𝑥 𝐶𝑠

km = 3.0566 μmol/L km = 2.3613 μmol/L
c) Eadie Hofstee c) Eadie Hofstee
𝑟 𝑟
𝑟 = 𝑟𝑚𝑎𝑥 + 𝑘𝑚 ( ) 𝑟 = 𝑟𝑚𝑎𝑥 + 𝑘𝑚 ( )
𝐶𝑠 𝐶𝑠

km = 2.8096 μmol/L km = 2.5083 μmol/
Therefore, Langmuir isotherm best fit the data with r = 0.9968 for withoutinhibitor.
2.18 The enzyme, cathepsin, hydrolyzes L-glutamyl-L-tyrosine to carbobenzoxy-L-glutamic acid

and L-tyrosine. It has been found (Frantz and Stephenson, J. Biol. Chem., 169, 359, 1947) that the
glutamic acid formed in the hydrolysis, inhibits (competitively) the progress of the reaction by
forming a complex with cathepsin. The course of the reaction is followed by adding tyrosine
decarboxylase which evolves CO2.
Substrate Inhibitor Rate of CO2 Generation

𝜇mole/mL 𝜇mole/mL 𝜇mole/mL.min
4.7 0 0.0434
4.7 7.57 0.0285
4.7 30.30 0.0133
10.8 0 0.0713
10.8 7.58 0.0512
10.8 30.30 0.0266
30.3 0 0.1111
30.3 7.58 0.0909
30.3 30.3 0.0581
Calculate (a) the value of Michaelis-Menten constants of the enzyme, Ks, and (b) the dissociation
constant of enzyme-inhibitor complex, KI.
600
y = 6.4106x + 329.1
500
R² = 0.9935
400
y = 6.5053x + 137.08
300 y =R²6.3732x
= 0.9986
+ 80.2
R² = 0.9993
200
100
0
0 5 10 15 20 25 30 35
a. @ I=0 @ I = 30.30
Rmax = 1/m = 0.1569 Rmax = 1/m = 0.1560
Ks = bRmax = 12.5839 KI = bRmax = 51.3368
@ I= 7.58
Rmax = 1/m = 0.1537
KI = bRmax = 21.0720
CHAPTER 2: ENZYME KINETICS ; ADDITIONAL PROBLEM
https://ww2.chemistry.gatech.edu/~lw26/bCourse_Information/3511/stud_comp/chap12_17.pdf
The following data were obtained for the reaction A ↔ B, catalyzed by the enzyme Aase. The
reaction volume was 1mL and the stock concentration of A was 5.0mM. Seven separate reactions
were examined, each containing a different amount of A. The reactions were initiated by adding
2.0µL of a 10µM solution of Aase. After 5 minutes, the amount of B was measured.
Reaction Volume of A Amount of B present
added(µL) at 5 minutes (nmoles)
1 8 26
2 10 29
3 15 39
4 20 43
5 40 56
6 60 62
7 100 71
(a) Calculate the initial velocity of each reaction (in units of µM.min-1)
(b) Determine the KM and Vmax of Aase from a Lineweaver-Burk plot.
(c) Calculate kcat.
SOL’N:
(a) νo = (26nmol/5min) / (1.0mL) x (103 mL/L) x (.001 µmol/1nmol) = 5.2 µM.min-1
Reaction νo(µM.min-1)
1 5.2
2 5.8
3 7.8
4 8.6
5 11.2
6 12.4
7 14.2
(b) Calculate [S] for each reaction
[A] = (.008mL)(5mM)(1mL) x (1000 µM/1 mM) = 40 µM
Reaction [S] µM (x) 1/[S] µM-1 ν(µM.min-1) (y) 1/ν (min-1/

µM)
1 40 0.025 5.2 0.192
2 50 0.02 5.8 0.172
3 75 0.0133 7.8 0.128
4 100 0.010 8.6 0.0116
5 200 0.005 11.2 0.089
6 300 0.0033 12.4 0.081
7 500 0.002 14.2 0.070
y-int = 1/vmax = 0.06

Vmax = 16 µM/min
x-int = -1/KM = 1/0.012
KM = 83 µM
(c) Calculate [E]T = (0.002mL)(10 µM Aase) / 1mL = 0.02 µM
Kcat = Vmax / [E]T = (16 µM/min) / 0.02 µM

Kcat= 800 /min
The statin drug lovastian helps lower cholesterol level by acts as competitive inhibitor on the
HMG-CoA reductase enzyme, which normally catalyzes an early step in the biosynthesis
pathway cholesterol
Required:
a) On a single graph, sketch the Michaelis-menten plot for HMG-CoA reductase in the presence
and absence of lovastatin, clearly labeling Km, and Vmax.
b) On a single graph, sketch the lineweaver-Burke (double reciprocal) plot for HMGCoA
reductase in the presence and absence of lovastatin. Clearly indicating how you could determin
Km, kcat and Vmax.
Solution:
a)
b)
Pesticide inhibition on enzyme has been reported, which caused the enzyme activity to reduce.
The collected data with and without inhibition are presented below. Determine the type of
inhibition and the KI for the inhibitor.
[S], M 3.30×10-4 5.00×10-4 6.70×10-4 1.65×10-3 2.21×10-3
Rate [I=0], 56 71 88 124 149
M/min×103
Rate
[I=20nM], 37 47 61 103 125
M/min×103
1 𝐾𝑚 1 1
Assuming Lineweaver-Burk Equation: =𝑉 +𝑉
𝑉 𝑚𝑎𝑥 𝑆 𝑚𝑎𝑥
[I=0]: Line 1: y=4.3200×10-9x+5.0033×10-6

1 𝐾𝑚
=5.0033×10-6 =4.3200×10-9
𝑉𝑚𝑎𝑥 𝑉𝑚𝑎𝑥
𝑉𝑚𝑎𝑥 = 199,868.0871 M/min 𝐾𝑚 = 8.6343×10-4 M
[I=20nM]: Line 2: y=7.4605×10-9x+5.1690×10-6

1 𝐾𝑚 𝑎𝑝𝑝
=5.1690×10-6 =7.4605×10-9
𝑉𝑚𝑎𝑥 𝑉𝑚𝑎𝑥
𝑉𝑚𝑎𝑥 = 193,461.0176 M/min 𝐾𝑚 𝑎𝑝𝑝 = 1.4433×10-3 M
𝐼 20×10−9 𝑀
𝐾𝑚 [1 + 𝐾 ] = 𝐾𝑚 𝑎𝑝𝑝 ; 8.6343×10-4 M [1 + ] =1.4433×10-3 M
𝐼 𝐾𝐼
𝐾𝐼 = 2.9780×10-8 M
Type of Inhibition: COMPETITIVE

𝐾𝐼 = 2.9780×10-8 M
A certain reaction has an activation energy of 125 kJ/mol. The rate is 0.33/s at 55ºC. Determine
the value of the specific rate constant at 100 ºC.
GIVEN: Ea=125 kJ/mol
@T1=55 ºC ; K55 ºC = 0.33
REQUIRED: @T2=100 ºC ; K100 ºC = ?
SOLUTION:
−𝐸𝑎
K =A𝑒 𝑅𝑇
@T1=55 ºC:
−125 𝑘𝐽/𝑚𝑜𝑙
8.314𝑘𝑗
(55+273)𝐾
0.33= A𝑒 𝑘𝑚𝑜𝑙.𝐾
A=2.6099×1019
@T2=100 ºC:
−125 kJ/mol
8.314kj
19 (100+273)K
K100ºC = (2.6099×10 )e kmol.K
K100ºC =82.8182/s
The enzyme carboxypeptidase catalyzes the hydrolysis of peptides. The following results were
obtained when the rate of enzymolysis of CBGP was monitored without inhibition at [CBGP]0=
0.713 mol/dm3.
𝐶𝐵𝐺𝑃 𝑚𝑜𝑙 1.25 3.84 5.81 7.13
,
10−2 𝑑𝑚3
𝑚𝑜𝑙 0.398 0.649 0.859 1.00
Rate, 𝑑𝑚3 .𝑠
When 2.0×10-3 mol/dm3 phenyl butyrate ion was added to the solution, the results were:
𝐶𝐵𝐺𝑃 𝑚𝑜𝑙 1.25 2.50 4.00 5.50
,
10−2 𝑑𝑚3
𝑚𝑜𝑙 0.172 0.301 0.344 0.548
In a separate experiment, the effect of 5.0×10-2 mol/dm3 benzoate ion was monitored and the
results were:
𝐶𝐵𝐺𝑃 𝑚𝑜𝑙 1.75 2.50 5.00 10.00
,
10−2 𝑑𝑚3
𝑚𝑜𝑙 0.183 0.201 0.231 0.246
Determine the type of inhibition and KI for phenyl butyrate and benzoate ion.
SOLUTION:
1 𝐾𝑚 1 1
Assuming Lineweaver-Burk Equation: =𝑉 +𝑉
𝑉 𝑚𝑎𝑥 𝑆 𝑚𝑎𝑥
Line 1: without inhibition

y=0.8126+0.0216x
𝑉𝑚𝑎𝑥 = 1.2306 mol/dm3.s
𝐾𝑚 = 0.0266 mol/dm3
Line 2: with phenyl butyrate ion
y=1.0158+0.0601x
𝐾𝑚 = 0.0592 mol/dm3
Line 3: with benzoate ion
y=3.7517+0.0300x
𝐾𝑚 = 8.0228 mol/dm3
TYPE OF INHIBITION:
 Phenyl Butyrate Ion: COMPETITIVE
 Benzoate Ion: UNCOMPETITIVE
FOR PHENYL BUTYRATE ION:

𝐼 2.0×10−3
𝐾𝑚 [1 + 𝐾 ] = 𝐾𝑚 𝑎𝑝𝑝 ; 0.0266 [1 + ] = 0.0592
𝐼 𝐾𝐼
𝐾𝐼 = 1.6369×10-3 mol/dm3
FOR BENZOATE ION:

𝑉𝑚𝑎𝑥 1.2306
𝑉𝑚𝑎𝑥 𝑎𝑝𝑝 = 𝐼 ; 0.2665= 5.0×10−2
1+ 1+
𝐾𝐼 𝐾𝐼
𝐾𝐼 = 0.0138 mol/dm3
CHAPTER 6: CELL KINETICS AND FERMENTER DESIGN
6.2 Aiba et al (1968) reported the results of a chemostat study on the growth of specific strain of
baker’s yeast as shown in the following table. The inlet stream of the chemostat did not contain
any cells or products.
Dilution Inlet Steady-state Steady-state Steady-state
Rate Glucose Conc. Glucose Conc. Ethanol Conc. Cell Conc.
D, hr-1 Cs, g/l Cs, g/l Cs, g/l Cs, g/l
0.084 21.5 0.054 7.97 2.00
0.100 10.9 0.079 4.70 1.20
0.160 21.2 0.138 8.57 2.40
0.198 20.7 0.186 8.44 2.33
0.242 10.8 0.226 4.51 1.25
a. Find the rate equation for cell growth.

b. Find the rate equation for product (ethanol) formation.
Given:
*Data refer to table
Required: rx, rp
Solution:
For a chemostat:
1
D=µ=τ
a. Assume Monod equation for cell growth
µ max Cs
µ= Cs + Ks
Linear form:
1 1 Ks 1
= µmax + µmax Cs
µ
Applying Linear Regression

µ max = 0. 4805 /hr
Ks = 0.2684 g/L
0.4805 CsCx
rx = 0.2684 + Cs
* The inlet stream of chemostat does not contain
b. For product formation any cells or product
dCx 1 dCx 2.00
rp= dt = Y x/p dt Yx/p = 7.97 = 0.2509
1.20
dCx µ max Cs Yx/p = = 0.2553
but = µCx = 4.70
dt Cs + Ks
2.40
1 µ max Cs
Yx/p = = 0.2800
8.57
rp = Y x/p Cs + Ks
2.33
Yx/p = = 0.2761
8.44
Δx
Yx/p = ΔP 1.25
Yx/p = =0.2772
4.51
Yx/p ave= 0.2679
1 0.4805 CsCx
rp = 0.2679 0.2684 + Cs
1.7936 CsCx
rp = 0.2684 + Cs
6.3 Andrews (1968) proposed the following model for the growth of microorganisms utilizing
inhibitory substrates.
µmax
µ= Ks Cs
1+ +
Cs KI
Assume that a chemostat study was performed with a microorganism. The volume of the fermenter
content was 1 L. The inlet stream was sterile. The flow rate and inlet substrate concentration were
varied and the steady-state concentration of glucose in the fermenter was measured and recorded
as shown in the table (the data are arbitrary).
a. Determine the kinetic parameters (µmax, KS, and KI) of this microorganism.
b. b. If the cell yield, YX/S, is 0.46 g/g, what is the steady-state cell concentration when the
flow rate is 0.20 L/h?
c. Andrews concluded in his paper that the primary result of substrate inhibition in a
continuous culture may be process instability. Explain what might happen if you suddenly
increase the substrate concentration from 30 to 60 g/L and why.
Flow Inlet Steady-state
Rate Glucose Conc. Glucose Conc.
F, l/hr-1 Cs, g/l Cs, g/l
0.20 30 0.5
0.25 30 0.7
0.35 30 1.1
0.50 30 1.6
0.70 30 3.3
0.80 30 10
0.50 60 30
0.60 60 22
0.70 60 15
Given:
*Data refer to table
Required:
a.) µmax, KS, and KI
b.) CX when T= 0.20L/h
c.) What happen if Csi is increased from 30 to 60 g/L
Solution:
a.
Ks Cs
1 1+ +
Cs KI
=
µ µmax
1 1 Cs Ks 1
= µmax ( 1 + ) + µmax Cs
µ KI
@ Csi = 30 g/l
1 30
(1+ ) = 0.8942371281
µmax KI
Ks
= 2.081776252
µmax
@ Csi = 60 g/l
1 60
(1+ ) = 2.49718382
µmax KI
Ks
= −16.47465438
µmax
1 0.8942371281
= 30
µmax 1+
KI
0.8942371281 60
30 (1+ ) = 2.49718382
1+ KI
KI
KI = -13.26329334
µ max= -1.41109489
Ks @ 30 g/l = -2.937
Ks @ 60 g/l = 23.2473
b.
@ F = ).20 L/h
Cx = YX/s (Csi – Cs)
Cx=0.46 ( 30 – 0.5 )
Cx = 13.57 g/l
c. If the initial substrate concentration is suddenly increased, the value of Ks changes.
6.6 The growth rate of E. coli in synthetic medium can be expressed by Monod kinetics as
0.935𝐶𝑠 𝐶𝑥
𝑟𝑥 = [g/L hr]
0.71+ 𝐶𝑠
where Cs is the concentration of a limiting substrate, glucose. You are going to cultivate E. coli
in a steady-state CSTF (working volume: 10 L) with a flow rate of 7 L/hr. The initial substrate
concentration is 10 g/L and the cell yield constant (YX/S) is 0.6. The feed stream is sterile.
a. What will be the doubling time and the division rate of the cells in the CSTF?
b. What will be the cell and substrate concentrations of the outlet stream?
c. If you connect one more 10-L CSTF to the first one, what will be the cell and substrate
concentrations in the second fermenter?
d. If you increase the flow rate from 7 to 10 L/hr for these two fermenters connected in series,
what will happen and why? Make a recommendation to avoid the problem if there is any.
Given:
0.935𝐶𝑠 𝐶𝑥
𝑟𝑥 = 0.71+ 𝐶𝑠
V = 10 L CSi = 10 g/L
F = 7 L/hr YX/S = 0.6
Sterile feed, Cxi = 0
Required:
a. td , δ
b. CS, CX
c. CS2, CX2
d. result
Solution:
1 1 𝐹 7 𝐿/ℎ𝑟
a. = ;𝐷= = = 0.7 ℎ𝑟 −1
𝐷 𝜇 𝑉 10 𝐿
µ = D = 0.7 hr-1
ln 2 ln 2
𝑡𝑑 = = = 0.9902 ℎ𝑟 td = 0.9902 hr
𝜇 0.7 ℎ𝑟 −1
1 1
𝛿=𝑡 = = 1.0099 ℎ𝑟 −1 δ = 1.0099 hr-1
𝑑 0.9902 ℎ𝑟
𝑉𝑑𝐶𝑥 𝑉𝑑𝐶𝑥
b. 𝐹𝐶𝑥𝑖 − 𝐹𝐶𝑥 + 𝑉𝑟𝑥 = 𝑏𝑢𝑡 =0
𝑑𝑡 𝑑𝑡
𝐹 (𝐶𝑥𝑖 − 𝐶𝑥 ) = −𝑉𝑟𝑥
𝐹 𝑟𝑥 0.935 𝐶𝑠 𝐶𝑥
= = (0.71+ 𝐶𝑠 )(𝐶𝑥 − 𝐶𝑥𝑖 )
𝑉 𝐶𝑥 − 𝐶𝑥𝑖
𝐹 0.935 𝐶𝑠
= (0.71+ 𝐶𝑠 )
𝑉
7 𝐿/ℎ𝑟 0.935 𝐶𝑠
= (0.71+ 𝐶𝑠 )
10 𝐿
Cs = 2.1149 g/L
𝐶𝑥 = 𝑌𝑥/𝑠 (𝐶𝑠𝑖 − 𝐶𝑠 )
𝐶𝑥 = 0.6(10𝑔/𝐿 − 2.1149𝑔/𝐿)
Cx = 4.7311 g/L
c. optimize CSTF
𝐾𝑠 +𝐶𝑠𝑖 0.71+10
𝛼 = √ = √ = 3.8839
𝐾𝑠 0.71
𝛼
𝐶𝑥1 = 𝐶𝑥,𝑜𝑝𝑡 = 𝑌𝑥/𝑠 𝐶𝑠𝑖 𝛼+1
𝑔 3.8839
= 0.6 (10 𝐿 ) 3.8839+1 = 4.7715 𝑔/𝐿
𝐶𝑠𝑖
𝐶𝑠1 = 𝐶𝑥,𝑜𝑝𝑡 = 𝛼+1
10 𝑔/𝐿
= = 2.0476 𝑔/𝐿
3.8839+1
𝐹 0.935𝐶𝑠2 𝐶𝑥2
= (0.71+ 𝐶𝑠2 ) (𝐶𝑥2 −𝐶𝑥1 )
𝑉
7 𝐿/ℎ𝑟 0.935𝐶𝑠2 𝐶𝑥2

= (1)
10 𝐿 (0.71+ 𝐶𝑠2 ) (𝐶𝑥2 −𝐶𝑥1 )
𝐶𝑥2 −𝐶𝑥1
𝑌𝑥/𝑠 = 𝐶𝑠1 −𝐶𝑠2
𝐶𝑥2 −𝐶𝑥1
0.6 = (2)
𝐶𝑠1 −𝐶𝑠2
Solve equations (1) 𝑎𝑛𝑑 (2) simultaneously
CS2 = 0.1214 g/L CX2 =5.9272 g/L

d. F = 10 L/hr
𝑉
𝜏𝑚 = 𝐹
For the second condition:

10 𝐿
𝜏𝑚 = = 1 ℎ𝑟 −1
10 𝐿/ℎ𝑟
𝜏𝑚 𝜇𝑚𝑎𝑥 = (1ℎ𝑟 −1 )(0.935) = 0.935

0.935 < 1
‫ ؞‬All the cells in the fermenter will be washed out if the flow rate is increased to 10 L/hr.
The flow rate should be less than the volume of the fermenter to avoid the wash out of the
cells formed.
6.7 Suppose that the growth rate of a microorganism can be expressed as the following equation:
rx = μmax (1 – e -CS/KS)CX
where μmax = 0.365 hr -1 and Ks = 6.8 g/L. The cell yield YX/S is found to be 0.45. If you cultivate
this microorganism in a 10-L CSTR with the flow rate of 2.8 L/hr, what will be the steady-
state cell concentration of the outlet stream? The substrate concentration of the inlet stream is
13 g/L. The inlet stream is sterile.
Given: rx = μmax (1 – e -CS/KS)CX

μmax = 0.365 hr -1 and Ks = 6.8 g/L
YX/S = 0.45 CSi = 13 g/L
V = 10 L F = 2.8 L/hr
Required: cell concentration of the outlet stream, CS
Solution:
rx = μmax (1 – e -CS/KS)CX
CX = YX/S(CSi – CS) = 0.45(13 g/L – CS)
rx = μCX
μ = F/V = 2.8 L/hr /10 L = 0.28 hr-1
0.28(0.45)(13 – CS) = 0.365(1 – e -CS/6.8)(0.45)(13 – CS)
CS = 9.9093 g/L
6.9 Suppose you have an organism that obeys Monod equation:
𝑑𝐶𝑥 𝜇𝑚𝑎𝑥 𝐶𝑠 𝐶𝑥
=
𝑑𝑡 𝐾𝑠 + 𝐶𝑠
where µmax = 0.5 hr -1 and Ks = 2 g/L
The organism is being cultivated in a steady-state CSTF, where F = 100L/hr, Csi = 50 g/L, and
Yx/s = 0.5
a. What size vessel will give the maximum total rate of cell production?
b. What are the substrate and cell concentrations of the optimum fermenter in part (a)?
c. If the exiting flow from the fermenter in part (a) is fed to a second fermenter (CSTF),
what should be the size of the second fermenter to reduce the substrate concentration
to 1 g/L?
d. If the exiting flow from the first fermenter in part (a) is fed to a second fermenter
whose size is the same as the first, what will be the cell and substrate concentrations
leaving the second fermenter?
Given:
Csi = 50 g/L Ks = 2 g/L
Yx/s = 0.5 F = 100 L/hr
µmax = 0.5/hr
Cs2
Cx2
Required:
a. V
b. Cx, opt and Cs, opt
c. V
d. Cs2 and Cx2
Solution:
𝐾𝑠 +𝐶𝑠𝑖 2+50
a. 𝛼 = √ = √ = 5.0990
𝐾𝑠 2
𝛼 5.0990
𝜏𝑚,𝑜𝑝𝑡 = 𝜇 = 0.5 (5.0990−1) = 2.4879 ℎ𝑟
𝑚𝑎𝑥 (𝛼−1)
𝑉 = (𝐹)( 𝜏𝑚,𝑜𝑝𝑡 ) = (100 𝐿⁄ℎ𝑟 )(2.4879 ℎ𝑟) = 248.7922 𝐿 V = 248.7922 L

𝛼 50𝑔 5.0990 𝑔
b. 𝐶𝑥,𝑜𝑝𝑡 = 𝑌𝑥/𝑠 𝐶𝑠𝑖 𝛼+1 = (0.5) ( ) (5.0990+1) = 20.9010
𝐿 𝐿
𝐶𝑠𝑖 50 𝑔/𝐿 𝑔
𝐶𝑠,𝑜𝑝𝑡 = = = 8.1980
𝛼 + 1 5.0990 + 1 𝐿
Cx, opt = 20.9010 g/L

Cs, opt = 8.1920 g/L
c. Cs2 = 1 g/L
𝑔 𝑔
𝐶𝑥2 = 𝑌𝑥/𝑠 (𝐶𝑠,𝑜𝑝𝑡 − 𝐶𝑠2 ) + 𝐶𝑥,𝑜𝑝𝑡 = (0.5)(8.1980 − 1) + 20.9010 = 24.5
𝐿 𝐿
100𝐿 2𝑔 24.5𝑔 20.9010𝑔
𝐹 (𝐾𝑠 +1)(𝐶𝑥2 −𝐶𝑥,𝑜𝑝𝑡 ) ( )( +1)( − )
ℎ𝑟 𝐿 𝐿 𝐿
𝑉= = 24.5𝑔 1𝑔 = 88.1393 𝐿
𝜇𝑚𝑎𝑥 𝐶𝑥,𝑜𝑝𝑡 𝐶𝑠2 (0.5/ℎ𝑟)( )( )
𝐿 𝐿
V = 88.1393 L
𝑔
d. 𝐶𝑥2 = 𝑌𝑥/𝑠 (𝐶𝑠,𝑜𝑝𝑡 − 𝐶𝑠2 ) + 𝐶𝑥,𝑜𝑝𝑡 = (0.5)(8.1980 − 𝐶𝑠2 ) + 20.9010 (1)
𝐿
100𝐿 0.5
𝐹 𝜇𝑚𝑎𝑥 𝐶𝑠2 𝐶𝑥2 ( )(𝐶𝑠2 )(𝐶𝑥2 )
= (𝐾 ℎ𝑟
= 2𝑔
ℎ𝑟
(2)
𝑉 𝑠 +𝐶𝑠2 )(𝐶𝑥2 −𝐶𝑥1 ) 248.7922 𝐿 (
𝐿
+𝐶𝑠2 )(𝐶𝑥2 −𝐶𝑥1 )
Solving for equations 1 and 2 simultaneously:
Cs2 = 0.2932 g/L

Cx2 = 24.8534 g/L
6.10. You are going to cultivate yeast, Saccharomyces Cerevisiae, by using a 10m3 fermenter your
company owns. You want to find out the amount of ethanol the fermenter can produce. Therefore,
a chemostat study was carried out and the Monod kinetic parameters. For the microorganism
grown in the glucose medium at 30oC, pH= 4.8, were found to be Ks= 0.025 g/L and umax= 0.25h-
1
. The ethanol yield (Yp/s) is 0.44 (g/g) and cell yield (YX/S) is 0.019 (g/g). The inlet substrate
concentration is 50 g/l.
a. What flowrate will give the maximum total ethanol production in the continuous fermenter and
what is the maximum ethanol production rate?
b. If you want to convert 95% of the incoming substrate, what must be the ethanol production rate
for the continuous fermenter?
c. If you have two 5m3 fermenters instead of one 10m3 fermenter, what is your recommendation
for the use of these fermenters to convert 95% of the incoming substrate? Would you recommend
connecting two fermenters in series to improve the productivity? Why or why not?
Given: V=10m3; Ks= 0.025 g/L; umax= 0.25h-1; Yp/s= 0.44; YX/S= 0.019 g/g; Cso= 50 g/l
Required:
(a) F
(b) F @ 95% Cso conversion
Solution:
0.025𝑔 50𝑔
𝐾𝑠+𝐶𝑠𝑜 +
(a) α= √ =√ 0.025𝑔/𝐿
𝐿 𝑙
=44.73
𝐾𝑠
𝐶𝑠𝑜
Cs= CS opt =𝛼+1= 50gL-1/ (44.73 +1) = 1.0934g/L
D= umax.CS/ (Ks+Cs)= 0.25h-1(1.0934g/L)/ (0.025 g/L+1.0934g/L)= 0.2444/h
F= DV= 0.2444/h (10m3)= 2.444 m3/h
(b) For 95% substrate conversion Cs= 0.05Cso = 4.75g/l.

0.25 𝑔
( )(4.75 )
ℎ 𝑙
D= umax.CS/ (Ks+Cs) = 𝑔 𝑔 = 0.0249 /h
(0.025 )+(4.75 )
𝑙 𝑙
F= DV= (0.0249/h) (10m3) = 0.2490 m3/h
(c) Cxo= YX/S(Cso- CS)= 0.019 (50- 4.75)g/L= 0.8598
g/L
𝛼 44.73
Cx1= YX/SCso(𝛼+1)= (0.019)(50)(4.573)=
0.9292g/L
𝐶𝑠𝑜 50g/l
Cs1= 𝛼+1= (44.73+1)= 1.0934g/L
D1= umax.CS/ (Ks+Cs) = 0.25h-1(1.0934g/L)/ (0.025
g/L+1.0934g/L)= 0.2444/h
F1= D1V1= 0.2444 h-1( 5m3)=1.2220 m3/h
𝛼
Cx2= YX/SCs1( )= (0.019)( 1.0934g/L)=0.0208g/L
𝛼+1
𝐶𝑠1 1.0934
Cs2= 𝛼+1= = 0.0239 g/L
45.73
0.0208−0.9292
YX/S= 0.0239−1.0934= 0.8494
Therefore; a two- 5 cubic meters of fermenters in series will increase the Growth yield
for 95% Ethanol conversion
CHAPTER 6: CELL KINETICS AND FERMENTER DESIGN ; ADDITIONAL PROBLEM
During the growth of E. coli in a batch reactor, the pattern can be modelled using the
Monod expression of the form;
µ𝑚𝑎𝑥
µ=
𝐾𝑆 + 𝑆
Where:
µ = specific growth rate
µmax =maximum specific growth rate
Ks = Monod constant
S = substrate concentration
For the above process, show how the time required to reach the maximum population of
cells can be estimated.
If the concentration of cells at the start of the exponential growth is 0.08g/L and the
essential substrate concentration is 36g/L. Find the time where the maximum population
of cells is reached given that ¹max = 0:55h¡1, Ks = 1:2g/L and k = 1:4gsubstrate/(gcells¢h)
Sol’n:
Substrate consumption during the growth can be described by;
𝑑𝑆
= −𝑘𝑋
𝑑𝑡
and the cell growth is given by;

𝑑𝑋
= µ𝑋
𝑑𝑡
where
1 𝑑𝑋
X = µ 𝑑𝑡
𝑑𝑆
Substitute X into 𝑑𝑡 and integrating gives;
𝑑𝑆 1 𝑑𝑋
= −𝑘( )
𝑑𝑡 µ 𝑑𝑡
𝑘
∫ 𝑑𝑆 = − ∫ 𝑑𝑋
µ
But
µ𝑚𝑎𝑥
µ=
𝐾𝑆 + 𝑆
hence;
𝑘
∫ 𝑑𝑆 = − µ ∫ 𝑑𝑋
𝑚𝑎𝑥
𝐾𝑆 + 𝑆
rearranging gives;
0 𝑋𝑆
𝑆 𝑘
∫ 𝑑𝑆 = − ∫ 𝑑𝑋
𝑆0 𝐾𝑆 + 𝑆 µ𝑚𝑎𝑥 𝑋0
Integrating both sides leads to;
µ𝑚𝑎𝑥 𝐾𝑆
𝑋𝑆 = 𝑋0 + (𝑆0 + 𝐾𝑆 𝑙𝑛 )
𝑘 𝐾𝑆 + 𝑆0
The average can is determined by dividing the speci¯c growth rate with the total
substrate in the reactor, which gives;
0
∫𝑆 µ𝑑𝑆
0
µ𝑎𝑣𝑔 = 0
∫𝑆 𝑑𝑆
0
0 𝑆
∫𝑆 𝐾 + 𝑆 𝑑𝑆
0 𝑆
µ𝑎𝑣𝑔 = 0
∫𝑆 𝑑𝑆
0
µmax[𝑆0 + 𝐾𝑠 𝑙𝑛 𝐾𝑆
]
𝐾𝑆 +𝑆0
µ𝑎𝑣𝑔 =
𝑆0
Therefore, the new cell growth expression should be in the form of;
𝑑𝑋
= µ𝑎𝑣𝑔 𝑋
𝑑𝑡
and integrating,
𝑡 𝑋𝑆
1 1
∫ 𝑑𝑡 = ∫ 𝑑𝑋
0 µ𝑎𝑣𝑔 𝑋0 𝑋
gives;
1 𝑋𝑆
𝑡= 𝑙𝑛
µ𝑎𝑣𝑔 𝑋0
substituting µavg previously lead to;
𝑆0 𝑋𝑆
𝑡= 𝑙𝑛
µ𝑚𝑎𝑥 𝐾 𝑋0
µ𝑚𝑎𝑥 [𝑋0 + (𝑆0 + 𝐾𝑆 𝑙𝑛 𝐾 +𝑆 𝑆 )]
𝑘 𝑆 0
Consider the amount of cell produced at the given substrate concentration:
Using;
µ 𝐾𝑆
𝑋𝑆 = 𝑋0 + 𝑚𝑎𝑥 (𝑆0 + 𝐾𝑆 𝑙𝑛 𝐾 +𝑆 )
𝑘 𝑆 0
0.55 1.2
𝑋𝑆 = 0.08 + (36 + 1.2 ln )
1.4 36 + 1.2
XS=12.604
and using the expression for µavg;
1.2
0.55[36 + 1.2𝑙𝑛 36 + 1.2]
µ𝑎𝑣𝑔 =
36
µ𝑎𝑣𝑔 = 0.487
thus, the time taken for the cell population to reach a maximum value;
1 𝑋𝑆
𝑡= 𝑙𝑛
µ𝑎𝑣𝑔 𝑋0
1 12.604
𝑡= 𝑙𝑛
0.487 0.08
t = 10.34 hrs.
Biochemical Engineering: A Textbook for Engineers, Chemists and Biologists
Shigeo Katoh and Fumitake Yoshida, 2009
Escherichia coli grows with a doubling time of 0.5 h in the exponential growth phase. (a) What
is the value of the specific growth rate? (b) How much time would be required to grow the cell
culture from 0.1 kg dry cell / m3?
Given: E. coli Req’d: a) µ

td=0.5 h b) t
Cn=10 Cno=0.1
Sol’n:
a)
ln 2
td = 𝜇
ln 2
µ= 0.5
µ=1.3868 h-1
b)
𝐶𝑛
ln(𝐶𝑛𝑜)=µ(t-to)
𝐶𝑛
ln( )
𝐶𝑛𝑜
t= µ
10
ln( )
.1
t=1.3863
t= 3.3219 h
Shigeo Katoh and Fumitake Yoshida, 2009
E. coli grows from 0.10 kg dry cell/m3 to 0.50 kg dry cell/m3 in 1 h.

1. Assuming the exponential growth during this period, evaluate the specific growth rate.
2. Evaluate the doubling time during the exponential growth phase.
3. How much time would be required to grow from 0.10 kg-dry cell/m3 to 1.0 kg dry cell/m3? You
may assume the exponential growth during this period.
Given: E. coli
Cn= 0.50 kg dry cell/m3 Cno=0.10 kg dry cell/m3
∆t=1 h
Req’d: a)µ
b) td
c) ∆t (from 0.1 to 1)
Sol’n:
a)
𝐶𝑛
ln
𝐶𝑛𝑜
µ=
∆𝑡
.5
ln(. 1)
µ=
1
µ=1.6094/h
b)
ln 2
td= 𝜇
td= ln 2 / 1.6094
td= 0.4307 h = 25.8412 min
c)
𝐶𝑛
ln
𝐶𝑛𝑜
∆𝑡 = µ
1
ln
.1
∆𝑡 =1.6094
∆t = 1.4307 h
Bioprocess Engineering Principles by Pauline M. Duran, pp. 364- 367
Steady state Concentration in a Chemostat

The Zymomonas mobilis cells are used for chemostat in a 60 m3 fermenter. The feed contains 12g/l
glucose; Ks= 0.2 g/L; Yxs=0.06 g/g; Ypx=7.7 g/g, umax=0.3/h; ms=2.2g/g.h;
Yps=Ypx.Yxs=0.46g/g.
a. What flowrate is required for a steady-state substrate concentration of 1.5 g/L?
b. At the flowrate of (a), what is the cell density?
c. At the flowrate of (a), what concentration of ethanol is produced?
Given: Ks= 0.2 g/L; Yxs=0.06 g/g; Ypx=7.7 g/g, umax=0.3/h; ms=2.2g/g.h;
Yps=Ypx.Yxs=0.46g/g.
Cs= 1.5 g/L, V= 60m3
Required:
a. F
b. Cx
c. Cp
Solution:
(a) D= umax.CS/ (Ks+Cs)= (0.3h-1)(1.5g/L)(0.2g/L + 1.5 g/L)= 0.26 h-1
F= DV= 0.26/h (60m3) = 15.6 m3h-1.
(b) When synthesis of the product is coupled with energy metabolism as for ethanol, Cx is
evaluated; therefore;
0.26
𝐷(𝐶𝑠𝑜−𝐶𝑆) ( )(12−1.5) 𝑔/𝑙
ℎ
Cx = 𝐷 = 0.26 = 0.42 g/L
+𝑚𝑠 ℎ +2.2
𝑌𝑥𝑠
0.06 ℎ
(c) Assuming ethanol is not present in the feed, Cpo=0. Steady-state product concentration is
3.4 𝑔
𝑞𝑝𝑥 ( )(0.42 )
ℎ 𝐿
Cp= 𝐶𝑝𝑜 + =0+ 0.26 = 5.5 g/L
𝐷 ( )
ℎ
Bioprocess Engineering Principles by Pauline M. Duran, pp. 364- 367
Substrate conversion and Biomass Productivity in a Chemostat

A 5 m3 fermenter is operated continuously with feed substrate concentration of 20 kg/m 3. The
microorganism cultivated in the reactor has the following characteristics: umax= 0.45/h; Ks= 0.8
kg/m3; Yxs= 0.55 kg/kg.
(a) What feed flow rate is required to achieve 90% substrate conversion?
(b) How does the biomass productivity at 90% substrate conversion compare with the
maximum possible?
Given: umax= 0.45/h; Ks= 0.8 kg/m3; Yxs= 0.55 kg/kg
Required: (a) F; (b) productivity comparison
Solution:
(d) For 90% substrate conversion Cs= 0.1 Cso = 2kg/m3.
0.45 𝑘𝑔
( )(2 )
ℎ 𝑚3
D= umax.CS/ (Ks+Cs) = 𝑘𝑔 𝑘𝑔 = 0.32 /h
(0.8 )+(2 )
𝑚3 𝑚3
F= DV= (0.32/h) (5m3) = 1.6 m3/h
(e) Assuming maintenance requirements and product formation are negligible:
0.8𝑘𝑔 0.32
𝐾𝑠𝐷 ( )( )
-1 3 𝑚3 ℎ
Qx= D (Cso- 𝑢𝑚𝑎𝑥−𝐷)Yxs = 0.32h (20kg.m - 0.45 0.32 )(0.55kg/kg)= 3.17 kg.m-3.h-1
( − )
ℎ ℎ
Maximum biomass @ Dopt
𝐾𝑆
Dopt= umax (1- √𝐾𝑆+𝐶𝑠𝑜 )= umax(1- α-1)
0.8𝑘𝑔
Dopt= 0.45h-1(1- √ 0.8𝑘𝑔𝑚3 𝑘𝑔 ) = 0.36h-1
+20
𝑚3 𝑚3
Maximum biomass productivity is determined with D= Dopt

𝐾𝑠𝐷
Qxmax= D (Cso- 𝑢𝑚𝑎𝑥−𝐷)Yxs
𝑘𝑔 0.36
(0.8 )( )
-3 𝑚3 ℎ
Dopt= 0.36/h (20kg.m - 0.45 0.32 ) (0.55 kg/kg) = 3.33 kg.m3.h-1
( − )
ℎ ℎ
𝟑.𝟏𝟕
Therefore, biomass productivity at 90% substrate conversion is 𝟑.𝟑𝟑x100 = 95% of the
theoretical maximum.
Bioprocess Engineering Principles by Pauline M. Duran, p. 375
Plug-Flow Reactor for immobilized enzymes:

Immobilised lactase is used to hydrolyze lactose in dairy waste to glucose and galactose. Enzyme
is immobilized in resin particles and packed into a 0.5 m3 column. The total effectiveness factor
for the system is close to unity; Km for the immobilized enzyme is 1.32 kg/m3; umax is 45 kg.m-
3 -1
h . The lactose concentration in the feed stream is 9.5 kg.m-3; a substance conversion 98% is
required. The column is operated with plug Flow for a total of 310 days a year.
a. At what flowrate should the reactor be operated?
b. How many tonnes of gluctose are produced a year?
Given: Cs= 0.02Cso= 0.19 kg.m-3; Cso= 9.5 kg.m-3; umax= 45 kg.m-3h-1; Km= 1.32 kg.m-3;
VPFR=0.5 m3
Required: a. F, b. mass glucose (Tons)
Solution:
(a) For 98% substrate onversion:
𝐾𝑚 𝐶𝑠𝑜 𝐶𝑠𝑜−𝐶𝑠
τ = 𝑢𝑚𝑎𝑥 𝑙𝑛 𝐶𝑠 + 𝑢𝑚𝑎𝑧 = 0.32 h
𝑉
F= τ = 1.56 m3/h
(b) The rate of lactose conversion is equal to the difference between inlet and outlet mass flow
rates of lactose= F(Cso-Cs)= 1.56 m3(0.5-0.19) kg/m3= 14.5 kg/h
𝑘𝑔 24ℎ 310𝑑 1𝑘𝑚𝑜𝑙 𝑘𝑚𝑜𝑙
Lactose converted= 14.5 . . . 342 𝑘𝑔 = 315
ℎ 1𝑑 1𝑦𝑟 𝑦𝑟
The enzyme reaction is:
Lactose+ H20 --) glucose + galactose

Therefore, from reaction stoichiometry, 315 kmol glucose are produced a year. The
molecular weight of glucose is 180; thus,
𝑘𝑚𝑜𝑙 180 𝑘𝑔 1𝑇 𝑻𝒐𝒏𝒏𝒆𝒔

Mass glucose= 315 . 1𝑘𝑚𝑜𝑙 . 1000𝑘𝑔 = 𝟓𝟔. 𝟕
𝑦𝑟 𝒚𝒓
Chemical Engineering by J. M. Coulson and J. F. Richardson
A continuous fermenter is operated at a series of dilution rates though at constant, sterile, feed
concentration, pH, aeration rate and temperature. The following data were obtained when the
limiting substrate concentration was 1200 mg/l and the working volume of the fermenter was 9.8
l. Estimate the kinetic constants Km,µm and kd as used in the modified Monod equation:
and also the growth yield coefficient Y.
Feed flowrate Exit substrate concentration Dry weight cell density

(l/h) (mg/l) (mg/l)
0.79 36.9 487
1.03 49.1 490
1.31 64.4 489
1.78 93.4 482
2.39 138.8 466
2.68 164.2 465
Solution
The accumulation = input − output + rate of formation,
which for the biomass gives:
V(dX/dt) = FX0 − FX + V(µmSX)/(Ks + S) − kdXV (equation 5.126)
and for the substrate:
V(dS/dt) = FS0 − FS − VµmSX/Y(Ks + S) (equation 5.127)
At steady state, dS/dt = 0.
Taking the dilution rate, D = F/V , then the balance for the substrate becomes:
S0 − S − {µmSX/[DY(Ks + S)]} = 0
or: X/D(S0 − S) = (KsY/µm)/S + Y/µm (i)

Similarly, for the biomass:
X0 − X + {µmSX/[D(Ks + S)]} − kdX/D = 0
Since the feed is sterile, X0 = 0 and:
DX = [µmSX/(Ks + S)] − kdX.

From the material balance for substrate:
(µmSX)/(Ks + S) = DY(S0 − S)
and substitution gives:

DX = DY(S0 − S) − kdX
or: (S0 − S)/X = kd/DY − 1/Y (ii)
From equation (i), it is seen that a plot of X/D(S0 − S) and 1/S will produce a straight line of
slope KsY/µm and intercept Y/µm.
From equation (ii), it is seen that a plot of (S0 − S)/X against 1/D will produce a straight line of
slope kd/Y and intercept 1/Y.
The data are calculated as follows:
Feed flowrate Exit 1/S 1/D

(F l/h) substrate (l/mg) (h)
(S mg/l)
0.79 36.9 0.0271 5.19 12.41 2.388
1.03 49.1 0.0204 4.05 9.51 2.349
1.31 64.4 0.0155 3.22 7.48 2.322
1.78 93.4 0.0107 2.40 5.51 2.296
2.39 138.8 0.0072 1.80 4.10 2.277
2.68 164.2 0.0061 1.64 3.66 2.228
The data are then plotted in Figure 5b from which:
KsY/µm = 170, Y/µm = 0.59, kd/Y = 0.0133 and 1/Y = 2.222.

6
5 2.4
4
3 slope, kd /Y = 0.0133/h
2.3
2
1
Y / µm = 0.59 X 1/Y = 2.222
0 slope, KsY/µm = 170 2.2
0 0.01 0.02 0.03 0 2 4 6 8 10 12
1/S (l/mg) 1/D (h)
Figure 5b. Graphical work
Thus:
yield coefficient, Y = (1/2.222) = 0.45 endogenous respiration coefficient,
µm = (0.45/0.59) = 0.76 h−1
and: Ks = (170 × 0.76/0.45) = 300 mg/l

Chemical Engineering by J. M. Coulson and J. F. Richardson
Two continuous stirred-tank fermenters are arranged in series such that the effluent of one forms
the feed stream of the other. The first fermenter has a working volume of 100 l and the other has
a working volume of 50 l. The volumetric flowrate through the fermenters is 18 h−1 and the
substrate concentration in the fresh feed is 5 g/l. If the microbial growth follows Monod kinetics
with µm = 0.25 h−1,Ks = 0.12 g/l, and the yield coefficient is 0.42, calculate the substrate and
biomass concentrations in the effluent from the second vessel. What would happen if the flow
were from the 50 l fermenter to the 100 l fermenter?
Solution
A material balance for biomass over the first fermenter, leads to the equation:
D1 = µ1 (equation 5.131)
where D1 is the dilution rate in the first vessel and µ1 is the specific growth rate for that vessel.
Assuming Monod kinetics to apply:
D1 = µmS1/(Ks + S1) (equation 5.132)
where S1 is the steady-state concentration of substrate in the first vessel, where:
S1 = D1Ks/(µm − D1) (equation 5.133)
If D1 = F/V1 = (18/100) = 0.18 h−1, then:
S1 = (0.18 × 0.12)/(0.25 − 0.18) = 0.309 g/l
Since the feed to this fermenter is sterile, X0 = 0 and from equation 5.134, Volume 3 the steady-
state concentration of biomass in the first vessel is given by:
X1 = Y(S0 − S1) (equation 5.134)
= 0.42(5 − 0.309) = 1.97 g/l
In a similar way, a mass-balance over the second vessel gives:
D2 = µ2X2/(X2 − X1) (equation 5.167)

where D2 is the dilution rate in the second vessel, µ2 is the specific growth rate in that vessel and
X2 the steady-state concentration of biomass.
The yield coefficient to the second vessel is then:
Y = (X2 − X1)/(S1 − S2)
where S2 is the steady-state concentration of substrate in that vessel.
Thus: X2 = X1 + Y(S1 − S2)
= 1.97 + 0.42(0.309 − S2)
= 2.1 + 0.42S2
Substituting this equation for X2 into equation 5.167, together with values for D2,µ2 and X1 leads
to a quadratic equation in S2:
0.
from which: and: X2 = 2.1 + (0.42 × 0.0113) = 2.1 g/l

S2 = 0.0113
g/l
When the tanks are reversed, that is with fresh feed entering
the 50 litre vessel, then the dilution rate for this vessel will be as before, 0.36 h−1, although the
critical dilution rate will now be:
Dcrit = µmS0/(Ks + S0) (equation 5.148)
= (0.25 × 5)/(0.12 + 5) = 0.244 h−1
This is lower than the dilution rate imposed and washout of the smaller vessel would take place.
The concentrations of substrate and biomass in the final effluent would eventually be those
attained if only the 100 litre vessel existed, that is:
concentration of biomass = 1.97 g/l
concentration of substrate .
CHAPTER 8: STERILIZATION
8.1 A fermenter containing 10 m3 of medium (25 °C) is going to be sterilized by passing saturated
steam (500 kPa, gage pressure) through the coil in the fermenter. The typical bacterial count
of the medium is about 3 × 1012 m-3, which needs to be reduced to such an extent that the
chance for a contaminant surviving the sterilization is 1 in 100. The fermenter will be heated
until the medium reaches 115 °C. During the holding time, the heat loss through the vessel is
assumed to be negligible. After the proper holding time, the fermenter will be cooled by
passing 20 m3/hr of 25 °C water through the coil in the fermenter until the medium reaches
40°C. The coils have a heat-transfer area of 40 m2 and for this operation the average overall
heat-transfer coefficient (U) for heating and cooling are 5,500 and 2,500 kJ/hr·m2·K,
respectively. The heat resistant bacterial spores in the medium can be characterized by an
Arrhenius coefficient (kdo) of 5.7 × 1039 hr-1 and an activation energy (Ed) of 2.834 × 105
kJ/kmol (Deindoerfer and Humphrey, 1959). The heat capacity and density of the medium are
4.187 kJ/kg·K and 1,000 kg/m3, respectively. Estimate the required holding time.
Given:
Vmedium = 10 m3 Theat = 115 °C Uheat = 5,500 kJ/hr·m2·K
Tmedium = 25 °C qwater = 20 m3/hr Ucool = 2,500 kJ/hr·m2·K
Psatd steam = 500 kPag Twater = 25 °C kdo = 5.7 × 1039 hr-1
N0 = 3 × 1012 m-3 Tcool = 40°C Ed = 2.834 × 105 kJ/kmol
N = 1/100 Acoil = 40 m2 Cp = 4.187 kJ/kg·K
ρ = 1,000 kg/m3
Required:
thold
Solution:
𝑁𝑜
∇ 𝑇 = ln 𝑁
3 ×1012
(10 m3 )
m3
∇T =ln 1 =35.6374
100
𝑎𝑡 𝑃𝑎𝑏𝑠 = 601.325 𝑘𝑃𝑎 ; 𝑇𝐻 = 432.0748 K (steam table)

Heating:
−𝑈𝐻 𝐴𝑡
𝑇ℎ𝑒𝑎𝑡 = 𝑇𝐻 + (𝑇𝑜 − 𝑇𝐻 ) 𝑒𝑥𝑝 𝐶𝑝 𝑀
𝑘𝑔⁄
M = 𝑉𝑚𝑒𝑑𝑖𝑢𝑚 𝜌 = (10𝑚3 ) (1000 𝑚3 ) = 10000 𝑘𝑔
𝑘𝐽
−(5500 )(40𝑚2 )𝑡
ℎ𝑟∙𝑚2 ∙𝐾
𝑇ℎ𝑒𝑎𝑡 = 432.0748𝐾 + (298𝐾 − 432.0748𝐾) 𝑒𝑥𝑝 𝑘𝐽
(4.187 )(10000𝑘𝑔)
𝑘𝑔∙𝐾
𝑇ℎ𝑒𝑎𝑡 = 432.0748𝐾 + (− 134.0748𝐾) exp(−5.2544 𝑡) at Theat = 388K ; t = 0.2117 hr

−𝐸𝑑
𝑡
∇ℎ𝑒𝑎𝑡= 𝑘𝑑𝑜 ∫0 𝑒 𝑅𝑇 𝑑𝑡
−2.834 × 105
5.7 × 1039 𝑡
∇ℎ𝑒𝑎𝑡= ∫0 𝑒 (8.318)(432.0748𝐾+(− 134.0748𝐾) exp(−5.2544 𝑡)) 𝑑𝑡 𝑎𝑡 t = 0.3894 hr
ℎ𝑟
∇heat = 0.6864
Cooling:
𝑈𝐴 𝑚𝑐 𝑡
𝑇𝑐𝑜𝑜𝑙 = 𝑇𝑐𝑜 + (𝑇𝑜 − 𝑇𝑐𝑜 ) exp{[1 − exp(𝑚 )] }
𝑐 𝐶𝑝 𝑀
2500 (40) 20000 𝑡

𝑇𝑐𝑜𝑜𝑙 = 298𝐾 + (388𝐾 − 298𝐾) exp{[1 − exp(20000(4.187))] }
10000
20000 𝑡
𝑇𝑐𝑜𝑜𝑙 = 298𝐾 + (90𝐾) exp{[− 2.3008] } at Tcool = 313K ; t = 0.3894 hr
10000
−𝐸𝑑
𝑡
∇𝑐𝑜𝑜𝑙= 𝑘𝑑𝑜 ∫0 𝑒 𝑅𝑇 𝑑𝑡
−2.834 × 105
5.7 × 1039 𝑡 20000 𝑡
∇𝑐𝑜𝑜𝑙= ℎ𝑟
∫0 𝑒 (8.318)(298𝐾+(90𝐾) exp{[− 2.3008] 10000 }) 𝑑𝑡 𝑎𝑡 t = 0.3894 hr
∇cool = 0.44
Holding:
∇hold = ∇T - ∇heat - ∇cool
∇hold = 35.6374 – 0.6864 – 0.44
∇hold = 34.511
𝐸 5.7 × 1039 −2.834 × 105
𝑘𝑑 = 𝑘𝑑𝑜 exp(𝑅𝑇𝑑 ) = exp ( (8.318)(388) ) = 41.6886
ℎ𝑟
𝛻ℎ𝑜𝑙𝑑
𝑡ℎ𝑜𝑙𝑑 = 𝑘𝑑
34.511
𝑡ℎ𝑜𝑙𝑑 = = 0.8278 thold = 0.8278
41.6886
8.2 A continuous sterilizer with a steam injector and a flash cooler will be employed to sterilize
medium continuously. The time for heating and cooling is negligible with this type of sterilizer.
The typical bacterial count of the medium is about 5x10^12 per m3, which needs to be reduced to
such an extent that only one organism can survive during the three months of continuous operation.
The heat resistant bacterial spores in the medium can be characterized by an Arrhenius coefficient
(kdo) of 5.7x10^39 per hr and an activation energy (Ed) of 2.834x10^5 kJ/kmol. The holding
section of the sterilizer will be constructed with 20m-long pipe with an inner diameter of 0.078 m.
Steam at 600 kPa(gage pressure) is available to bring the sterilizer to an operating temperature of
125 deg C. The physical properties of this medium at 125 degC are c= 4.187 kJ/kg , density= 1000
kg/m3 , and viscosity = 4kg/m.hr.
a. How much medium can be sterilized per hour if you assume ideal plug flow?
b. How much medium can be sterilized per hour if the effect of axial dispersion is considered?
Given:
No = 5x10^12 per m3 L=20m
N = 1/3months d = 0.078m
kdo = 5.7x10^39 per hr P = 600 kPag
Ed = 2.834x10^5 kJ/kmol
Required:
a. Q at ideal plug flow

b. Q with axial dispersion
Solution:
a. = ln (5x10^32)(Q)(24)(90) b. Q= 0.8 m3/hr (assumed)
1
= ln ((1.08x10^16)x Q) Nre= 3.264 x 10^3
= Kd Thold D/udt = 1.1 ; D=14.3648 m2/hr
Kd=Kdo e^(-E/RT) Npe= 233.0995
Kd= 363.3440 /hr kdL/u = 43.4047
u= Q at the graph:
(π/4) (0.078)^2 n/no= 9.8 x 10^-17
Which is almost equal to e^(-36.8587)
L= u Thold =20 m n/no= 9.8282 x 10^-17
Thold = ln((1.08x10^16)xQ)
363.3440
Q= 0.9421 m3/hr
= 36.8587
u= 197.1596 m/hr
Thold= 0.1014 hr
8.3 You need to design a filter for a 10,000-gallon fermenter that will be aerated at a rate of 535 ft
3
/min (at 200C and 1 atm). The bacterial count in the air is 80 per ft3. Average size of the bacteria
is 1 µm with density of 1.08g/cm3. You are going to use glass fibers (Dc = 15 µm) with packing
density α = 0.03. The cross-sectional area of the filter will be designed to give a superficial air
velocity υo of 5 ft/s.
a. What depth of the filter would you recommend to prevent contamination?
b. How is the answer in (a) change if υo is decreased to 1 ft/s? Explain the results.
Given:
Vt = 10,000 gal @20oC & 1atm
Q = 535 ft3/min ρ = 1.2 x10-3 g/cm3
Cno = 80/ft3 μ = 1.8x10-4
dp = 1 μm= 1x10-4 cm
Dc = 15 μm= 1.5x10-3 cm
α = 0.03
υo = 5 ft/s
Required:
a. B
b. B if vo is decreased to 1 ft/s
Solution:
5ft 100 𝑐𝑚
( )( ) 157.1134𝑐𝑚
s 3.28 𝑓𝑡
a. V= =
1−0.03 𝑠
𝑐𝑚
𝐷𝑐 𝑉𝜌 (1.5𝑋10−3 𝑐𝑚)(157.1134 )(1.2𝑥10−3 )
𝑠
𝑁𝑅𝑒 = = = 1.5711
𝜇 1.8𝑋10−4
𝜇 𝜋𝑀𝑤 1.8𝑥10−4 𝜋(29)

𝜆=( )√ =[ −3
] [√ ] = 6.4995𝑥10−6 𝑐𝑚
0.499𝜌 8𝑅𝑇 (0.499)(1.2𝑥10 ) 8(8.314𝑥107 )(293)
−1.10(1x10−4)
2𝜆 −1.10𝑑𝑝 2(6.4995𝑥10−6 )
𝐶𝑓 = 1 + (1.257 + 0.400𝑒 2𝜆 ) = 1 + −4
(1.257 + 0.400𝑒 2(6.4995𝑥10−6) ) = 1.16
𝑑𝑝 1x10
𝐶𝑓 𝜌𝑝 𝑑2 𝑝 𝜐 (1.1634)(1.08)(1x10−4 )2 (157.1134)
𝑁𝑠𝑡 = = = 0.4062
18𝜇𝐷𝑐 18(1.8x10−4 )(1.5𝑥10−3 )
𝑁𝑖𝑚𝑝 = 0.075𝑁𝑆𝑡 1.2 = 0.075(0.4062)1.2 = 0.0254
𝑑𝑝 1𝑥10−4
𝜅= = = 0.0667
𝐷𝑐 1.5𝑥10−3
1 𝜅(2 + 𝜅)
𝑁𝑖𝑛𝑡 = [(1 + 𝜅) ln(1 + 𝜅) −
2.002 − 𝑙𝑛𝑁𝑅𝑒 2(1 + 𝜅)
1 0.0667(2+0.0667)
= 2.002−ln(1.5711) [(1 + 0.0667) ln(1 + 0.0667) − 2(1+0.0667)
= 2.7466𝑥10−3
𝐶𝑓 𝜅𝑇 (1.16)(0.0667)(293)
𝒟𝐵𝑟 = = = 2.7729𝑥10−7
3𝜋𝜇𝑑𝑝 3𝜋(1.8𝑥10−4 )(1𝑥10−4 )
𝐷𝑐 𝜐 (1.5𝑥10−3 )(157.1134)
𝑁𝑃𝑒 = = = 8.4987𝑥105
𝒟𝐵𝑟 2.7729𝑥10−7
𝑁𝑑𝑖𝑓𝑓 = 1.3𝑁𝑃𝑒 −2/3 + 0.7𝜅 2 = 1.3(8.4987𝑥105 )−2/3 + 0.7(0.0667)2 = 3.2560𝑥10−3
𝜂𝑐 = 1 − (1 − 𝜂𝑖𝑚𝑝 )(1 − 𝜂𝑖𝑛𝑡 )(1 − 𝜂𝑑𝑖𝑓𝑓 )
= 1 − (1 − 0.0254)(1 − 2.7466𝑥10−3 )(1 − 3.2560𝑥10−3 ) = 0.0312
Assume T= 1min operation
𝐶𝑛 −4𝐵 𝛼
ln ( ) = ( )𝜂
𝐶𝑛𝑜 𝜋𝐷𝑐 1 − 𝛼 𝑐
1 −4𝐵 0.03
ln [ 80 𝑓𝑡3
] = 𝜋(1.5𝑥10−3 ) (1−0.03)(0.0312)
( 3)(535 )(1min)
𝑓𝑡 𝑚𝑖𝑛
B=13.0027 cm
b. if vo=1ft/s
1ft 100 𝑐𝑚
( )( ) 31.4307𝑐𝑚
s 3.28 𝑓𝑡
V= =
1−0.03 𝑠
𝑐𝑚
𝐷𝑐 𝑉𝜌 (1.5𝑋10−3 𝑐𝑚)(31.4307 )(1.2𝑥10−3 )
𝑠
𝑁𝑅𝑒 = = = 0.3143
𝜇 1.8𝑋10−4
𝜇 𝜋𝑀𝑤 1.8𝑥10−4 𝜋(29)

𝜆=( ) √ =[ ] [ √ ] = 6.4995𝑥10−6 𝑐𝑚
0.499𝜌 8𝑅𝑇 (0.499)(1.2𝑥10−3 ) 8(8.314𝑥107 )(293)
−1.10(1x10−4)
2𝜆 −1.10𝑑𝑝 2(6.4995𝑥10−6 ) 2(6.4995𝑥10−6 ) ) = 1.16
𝐶𝑓 = 1 + (1.257 + 0.400𝑒 2𝜆 ) = 1 + (1.257 + 0.400𝑒
𝑑𝑝 1x10−4
𝐶𝑓 𝜌𝑝 𝑑2 𝑝 𝜐 (1.1634)(1.08)(1x10−4 )2 (31.4307)
𝑁𝑠𝑡 = = = 0.0813
18𝜇𝐷𝑐 18(1.8x10−4 )(1.5𝑥10−3 )
𝑁𝑖𝑚𝑝 = 0.075𝑁𝑆𝑡 1.2 = 0.075(0.0813)1.2 = 0.0037
𝑑𝑝 1𝑥10−4
𝜅= = = 0.0667
𝐷𝑐 1.5𝑥10−3
1 𝜅(2 + 𝜅)
𝑁𝑖𝑛𝑡 = [(1 + 𝜅) ln(1 + 𝜅) −
2.002 − 𝑙𝑛𝑁𝑅𝑒 2(1 + 𝜅)
1 0.0667(2+0.0667)
= 2.002−ln(0.3143) [(1 + 0.0667) ln(1 + 0.0667) − 2(1+0.0667)
= 0.0043
𝐶𝑓 𝜅𝑇 (1.16)(0.0667)(293)
𝒟𝐵𝑟 = = = 2.7729𝑥10−7
3𝜋𝜇𝑑𝑝 3𝜋(1.8𝑥10−4 )(1𝑥10−4 )
𝐷𝑐 𝜐 (1.5𝑥10−3 )(31.4307)
𝑁𝑃𝑒 = = = 1.7001𝑥105
𝒟𝐵𝑟 2.7729𝑥10−7
𝑁𝑑𝑖𝑓𝑓 = 1.3𝑁𝑃𝑒 −2/3 + 0.7𝜅 2 = 1.3(1.7001𝑥105 )−2/3 + 0.7(0.0667)2 = 3.5378𝑥10−3
𝜂𝑐 = 1 − (1 − 𝜂𝑖𝑚𝑝 )(1 − 𝜂𝑖𝑛𝑡 )(1 − 𝜂𝑑𝑖𝑓𝑓 )

= 1 − (1 − 0.0037)(1 − 0.0043)(1 − 3.5378𝑥10−3 ) = 0.0115
Assume T= 1min operation
𝐶𝑛 −4𝐵 𝛼
ln ( ) = ( )𝜂
𝐶𝑛𝑜 𝜋𝐷𝑐 1 − 𝛼 𝑐
1 −4𝐵 0.03
ln [ 3 ]= −3
( )(0.0115)
80 𝑓𝑡 𝜋(1.5𝑥10 ) 1 − 0.03
( 3 ) (535 𝑚𝑖𝑛) (1min)
𝑓𝑡
B = 35.3237 cm
CHAPTER 8: STERILIZATION ; ADDITIONAL PROBLEM
Shigeo Katoh and Fumitake Yoshida , 2009
A medium is to be continuously sterilized at a flow rate of 2m3h-1 a sterilizer by direct steam

injection. The temperature of a holding section is maintained at 120°C, and the time for heating
and cooling can be neglected. The bacterial count of the entering medium, 2x1012m-3, must be
reduced to such an extent that only one organism can survive during 30 days of continuous
operation. The holding section of the sterilizer is a tube, 0.15 m in the internal diameter. The
specific death rate of bacterial spores in the medium is 121 h-1 at 120°C, the medium density ρ=950
kg m-3, the medium viscosity μ=1kgm-1h-1 . Calculate the required length of the holding section of
the sterilizer
(a) Assuming ideal plug flow.
(b) Considering the effect of the axial dispersion.
Given: Sterilization by steam injection

F=2 m3/h
Th=120°C
theating and tcooling is negligible
Cno=2x1012m-3
Cn= 1/1000 in 30 d
Kd=121 h-1
ρ=950 kg m-3
μ=1kgm-1h-1
Req’d: L a) ideal PFR

b) consider axial dispersion
Sol’n:
24h
𝑛 (2x1012m−3)(2m3h−1)(30 d)( )
d
a)ln ( ) = = 35.6
𝑛𝑜 1
Average holding time:
𝑛
ln( )
𝑛𝑜
𝜏hold= =0.294 h
𝐾𝑑
The average medium velocity in the holding section:

2
𝑣= = 113𝑚/ℎ
𝜋(0.0752 )
The required length of the holding section:
L= (113m/h) (0.294h)
L=33.2 m
b) The Reynolds number of the medium flowing through the holding tube is
0.15 × 113 × 950

𝑅𝑒 = = 1.64 × 104
1
From Figure 10.3 Pe is approximately 2.5 at

Re=1.53x104
EDz=(113 x 0.15)/2.5
EDz=6.8
Assuming a length of the holding section of 36 m, the Peclet number is given as

𝑣𝐿 113 ×36
𝑃𝑒 = 𝐸 = 6.8
Pe=598 and
121 ×36
Da= KdL/v= 113
From Figure 10.4,
the value of n/n0 is approximately 3.5x10-16, and almost equal to that required in this problem.
Thus, the length of the holding section should be 36 m.
L = 36 m
CHAPTER 8: STERILIZATION ; ADDITIONAL PROBLEM
Calculate the total degree of batch sterilization, Δtotal, for a liquid medium inside a bioreactor
vessel, which reaches maximum temperature 120 oC, and then cooled off. Assume that the liquid
medium contains spores of B. stearothermophilus, and the initial total number is N0 = 6 x 1012
spores. The temperature vs. time profile during batch sterilization is given below.
t (min) T1 (oC)
0 30
10 50
30 90
36 100
43 110
50 120
55 120
58 110
63 100
70 90
102 60
120 44
140 30
For spores of B. stearothermophilus:

k = 7.94 x 1038 exp[(-68.8 x 103)/RT] min-1
R = 1.98 cal/g mole oK
FIG. 4.8 Batch sterilization: k and T vs. t ; example calculation. Area under the curve k vs. t is
total degree of sterilization, Δtotal. [Adopted from S. Aiba, A.E. Humphrey and N.F. Millis.
“Media Sterilization”. In Biochemical Engineering, 2nd Ed., Academic Press, Inc., New York
(1973) 256].
Req’d: ΔTotal
Solution:
Examining Fig. 4.8, it is also evident that the values of k are a function of t [i.e. k = f (t)], ranging
between 0 to 34 min, and between 64 to 140 min. Therefore, the area under the curve k (min-1)
vs. t (min) is the graphical integration, which gives:
Δtotal = ln(N0/N) = 0140 kdt = 33.8

N = N0/exp(33.8) = 6x1012/4.77698x104 = 1.256x10-2
CHAPTER 9: AGITATION AND AERATION
9.1 Derive the relationship between the overall mass transfer coefficient for gas phase KG and the
individual mass-transfer coefficients, kL and kG. How can this relationship be simplified for
sparingly soluble gases?
Given:
a. Overall mass transfer coefficient for gas phase, KG
b. Individual mass transfer coefficients, kL and kG
Required:
a. Derive the relationship between KG, and kG and kL
b. How can the relationship be simplified for sparingly soluble gases?
Solution:
Expressing the molar flux in terms of overall driving force for mass transfer and an
overall mass transfer coefficient,
𝑁𝐴 = 𝐾𝐺 (𝑃𝐴,𝐺 − 𝑃 ∗𝐴 )
𝑁𝐴 = 𝐾𝐿 (𝑐 ∗𝐴 − 𝑐𝐴,𝐿 )
where KG is termed the “overall mass transfer coefficient based on gas phase driving
forces.” In an equivalent manner, we also can write
Using Henry’s Law,

𝑃𝐴,𝐺 = 𝑚𝑐 ∗𝐴
where m is the Henry’s law constant.
We can rewrite the first two equations as,

𝑁𝐴
𝑃𝐴,𝐺 − 𝑃𝐴,𝑖 =
𝑘𝐺
𝑁𝐴
𝑐𝐴,𝑖 − 𝑐𝐴,𝐿 =
𝑘𝐿
Substituting the equilibrium relationships,
𝑚𝑁𝐴
𝑃𝐴,𝑖 − 𝑃 ∗𝐴 =
𝑘𝐿
Adding the partial pressure differences we obtain
𝑁𝐴 𝑚𝑁𝐴 1 𝑚
𝑃𝐴,𝐺 − 𝑃𝐴,𝑖 + 𝑃𝐴,𝑖 − 𝑃 ∗𝐴 = 𝑃𝐴,𝐺 − 𝑃 ∗𝐴 = + = 𝑁𝐴 ( + )
𝑘𝐺 𝑘𝐿 𝑘𝐺 𝑘𝐿
Simplifying further,
1 1 𝑚
= +
𝐾𝐺 𝑘𝐺 𝑘𝐿
For sparingly soluble gases,

1
=0
𝑘𝐺
Therefore,
1 𝑚
=
𝐾𝐺 𝑘𝐿
9.2 Prove that Eq. (9.25) is the same with Eq. (9.26), and Eq. (9.27) is the same with Eq. (9.28)
Given:
a) Eq. (9.25) and Eq. (9.26)
b) Eq. (9.27) and Eq. (9.28)
Required:
a) Prove that
2
∆𝜌𝜇𝑐 𝑔 1/3
Eq. (9.25) 𝑘𝐿 = 0.31𝑁𝑆𝑐 −3 ( ) is the same as
𝜌2
Eq. (9.26) 𝑁𝑆ℎ = 0.31𝑁𝑆𝑐 1/3 𝑁𝐺𝑟 1/3
b) Prove that
2
2𝐷𝐴𝐵 ∆𝜌𝜇𝑐 𝑔 1/3
Eq. (9.27) 𝑘𝐿 = + 0.31𝑁𝑆𝑐 −3 ( ) is the same as
𝐷32 𝜌2
Eq. (9.28) 𝑁𝑆ℎ = 2.0 + 0.31𝑁𝑆𝑐 1/3 𝑁𝐺𝑟 1/3
Solution:
a.)
2
∆𝜌𝜇𝑐 𝑔 1/3
𝑁𝑆ℎ = 0.31𝑁𝑆𝑐 1/3 𝑁𝐺𝑟 1/3 𝑘𝐿 = 0.31𝑁𝑆𝑐 −3 ( )
𝜌2
Substituting Eq. (9.21), Eq. (9.22) and Eq. (9.23) into Eq. (9.26):
𝑘𝐿 𝐷32 𝜇𝑐 3 𝜌 𝑔∆𝜌
𝐷32
= 0.31( 𝜌 )1/3 ( 𝑐
)1/3
𝐷𝐴𝐵 𝑐 𝐷𝐴𝐵 𝜇𝑐2
𝐷32
𝜇𝑐 1/3 𝜌𝑐 1/3 𝑔1/3 ∆𝜌1/3 Dividing both sides by
𝑘𝐿 = 0.31 2/3 𝐷𝐴𝐵
𝜌𝑐 1/3 𝐷𝐴𝐵 −2/3 𝜇𝑐
𝜇𝑐 −1/3 𝜌𝑐 1/3 𝑔1/3 ∆𝜌1/3 Combining similar terms

𝑘𝐿 = 0.31
𝜌𝑐 𝐷𝐴𝐵 −2/3
1/3 1
𝜇𝑐 −1/3 𝜌𝑐 1/3 𝑔1/3 ∆𝜌1/3 −1/3 1/3

𝑘𝐿 = 0.31 Multiplying (
𝜇𝑐 𝜇𝑐
−2/3 )( )
−2/3
𝜌𝑐 1/3 𝐷𝐴𝐵 1 2/3
𝜌𝑐 𝜌𝑐
𝜇𝑐 −2/3 𝜇𝑐 1/3 𝑔1/3 ∆𝜌1/3

𝑘𝐿 = 0.31
𝜌𝑐 −2/3 𝐷𝐴𝐵 −2/3 𝜌2/3 𝑐
𝜇𝑐 −2 ∆𝜌𝜇𝑐 𝑔 1/3
𝑘𝐿 = 0.31( ) 3( )
𝜌𝑐 𝐷𝐴𝐵 𝜌𝑐 2
2 2
∆𝜌𝜇𝑐 𝑔 1/3 ∆𝜌𝜇𝑐 𝑔 1/3
𝑘𝐿 = 0.31𝑁𝑆𝑐 −3 ( ) 𝑘𝐿 = 0.31𝑁𝑆𝑐 −3 ( )
𝜌2 𝜌2
b.)
2𝐷𝐴𝐵
𝑁𝑆ℎ = + 0.31𝑁𝑆𝑐 1/3 𝑁𝐺𝑟 1/3 𝑘𝐿 = 2.0 +
𝐷32
2
− ∆𝜌𝜇𝑐 𝑔 1/3
0.31𝑁𝑆𝑐 3 ( 𝜌2 )
Substituting Eq. (9.21), Eq. (9.22) and Eq. (9.23) into Eq. (9.28):
3
𝑘𝐿 𝐷32 𝜇𝑐 𝐷32 𝜌𝑐 𝑔∆𝜌 1/3
= 2.0 + 0.31( 𝜌 )1/3 ( )
𝐷𝐴𝐵 𝑐 𝐷𝐴𝐵 𝜇𝑐2
𝐷32
𝐷 𝜇𝑐 1/3 𝜌𝑐 1/3 𝑔1/3 ∆𝜌1/3 Dividing both sides by
𝑘𝐿 = 2 𝐷𝐴𝐵 + 0.31 2/3 𝐷𝐴𝐵
32 𝜌𝑐 𝐷𝐴𝐵 −2/3
1/3
𝜇𝑐
𝐷 𝜇𝑐 −1/3 𝜌𝑐 1/3 𝑔1/3 ∆𝜌1/3 Combining similar terms

𝑘𝐿 = 2 𝐷𝐴𝐵 + 0.31
32 𝜌𝑐 𝐷𝐴𝐵 −2/3
1/3 1
𝐷 𝜇𝑐 −1/3 𝜌𝑐 1/3 𝑔1/3 ∆𝜌1/3

𝑘𝐿 = 2 𝐷𝐴𝐵 + 0.31 𝜇𝑐
Multiplying (
−1/3
𝜇𝑐
)(
1/3
)
32 𝜌𝑐 1/3 𝐷𝐴𝐵 −2/3 1 −2/3 2/3
𝜌𝑐 𝜌𝑐
𝐷𝐴𝐵 𝜇𝑐 −2/3 𝜇𝑐 1/3 𝑔1/3 ∆𝜌1/3

𝑘𝐿 = 2 + 0.31
𝐷32 𝜌𝑐 −2/3 𝐷𝐴𝐵 −2/3 𝜌2/3 𝑐
𝐷𝐴𝐵 𝜇𝑐 −2 ∆𝜌𝜇𝑐 𝑔 1/3
𝑘𝐿 = 2 + 0.31( ) 3( )
𝐷32 𝜌𝑐 𝐷𝐴𝐵 𝜌𝑐 2
2
𝐷𝐴𝐵 ∆𝜌𝜇𝑐 𝑔 1/3 𝐷𝐴𝐵
𝑘𝐿 = 2 + 0.31𝑁𝑆𝑐 −3 ( ) 𝑘𝐿 = 2 +
𝐷32 𝜌2 𝐷32
2
− ∆𝜌𝜇𝑐 𝑔 1/3
0.31𝑁𝑆𝑐 ( 3 )
𝜌2
9.4 A cylindrical tank (1.22m diameter) is filled with water to an operating level equal to the tank
diameter. The tank is equipped with four equally spaced baffles, the width of which is one tenth
of the tank diameter. The tank is agitated with a 0.36 m diameter, flat-blade disk turbine. The
impeller rotational speed is 4.43 rps. The air enters through an open ended tube situated below the
impeller and its volmetric flow rate is 0.0217 m3/s at 1.08 atm and 25 deg C.
density= 997.08 kg/m3 viscosity = 8.904 x 10^-4 kg/m-s
Calculate:
a. Power Requirement
b. Gas Hold-up
c. Sauter-mean Diameter
d. Interfacial area
e. Volumetric mass-transfer coefficient
Given:
DT = 1.22 m
W = 1/10 DT
DI = 0.36 m
N = 4.43 rps
Q = 0.0217 m3/s
Required:
a. Pm
b. H
c. D32
d. a
e. kla
Solution:
a. Nre = (997.08)(4.43)(.36)^2 d. Interfacial Area:
8.904 x 10^-4 a =6H/ D32
Nre = 642,915.034 > 10,000 a= 98.5447 /m
Pmo = 6 (997.08) (4.43)^3 (.36) ^5
= 3144.8862 W e. Volumetric mass-trans coeff.
Using equation 9.53 of James Lee Kl = 4.58x10^-4 m/s
Pm= 1341.3218 W Kla = 4.58x10^-4 (98.5447)
b. v= (π/4)(1.22)(1.22)^2 Kla = 0.0451 /s
v= 1.43 m3
Vs= (4x .0217) / π (1.22)^2

= 0.0186 m/s < 0.02 m/s
Using equation 9.48 of James Lee

H = 0.0790
c. Using equation 9.42
D32= 4.8132 x 10^-3 m
D32= 4.8132 mm
9.5 Estimate the volumetric mass-transfer coefficient kLa for the gas-liquid contractor described
in Problem 9.4 by using a correlation for kLa and compare the result with the experimental value.
Given:
Reactor volume, v= 1.43m3
Vs= 0.0186 m/s
PM= 1342 Watts
Required:
kLa (using equation 9.71 by James Lee)
% kLa compared with experimental value
Solution:
1342
kLa = 0.026( 1.43 )0.4 (0.0186)0.5 = 0.0548 s-1 Experimental
Estimated value= 0.0451 s-1

(0.0548−0.0451)𝑥 100%
% for volumetric mass-transfer coefficient = = 17.7007% error
(0.0548)
9.6 The power consumption by an agitator in an unbaffled vessel can be expressed as

𝑃𝑚𝑜 𝜌𝑁𝐷𝐼2
= 𝑓 ( )
𝜌𝑁 3 𝐷𝐼5 𝜇
Can you determine the power consumption and impeller speed of a 1,000-gallon fermenter
based on findings of the optimum condition from a one-gallon vessel by using the same fluid
system? Is your conclusion reasonable? Why or why not?
Given:
𝑃𝑚𝑜 𝜌𝑁𝐷𝐼2
= 𝑓 ( )
𝜌𝑁 3 𝐷𝐼5 𝜇
VP =1000 gallons
Vm=1 gallon
Required:
Can the power consumption and impeller speed of VP be determined on findings of the
optimum condition from Vm by using the same fluid system? Why?
Solution:
𝑉𝑃
= 1000
𝑉𝑚
The scale ratio is
𝐷𝐼,𝑃 1
= 1000 ⁄3 = 10
𝐷𝐼,𝑚
To achieve dynamic similarity, the three numbers for the prototype and model must be
equal
𝑃𝑚𝑜 𝑃𝑚𝑜
[ 3 5] = [ 3 5]
𝜌𝑁 𝐷𝐼 𝑃 𝜌𝑁 𝐷𝐼 𝑚
𝜌𝑁𝐷𝐼2 𝜌𝑁𝐷𝐼2
[ ] =[ ]
𝜇 𝑃 𝜇 𝑚
Using the same fluid for model and prototype, 𝜌𝑃 = 𝜌𝑚 ; 𝜇𝑃 = 𝜇𝑚
𝑁𝑃 3
(𝑃𝑚𝑜 )𝑝 = 105 [𝑃𝑚𝑜 ]𝑚 [ ]
𝑁𝑚
The equality of Reynold’s number requires
𝑁𝑃 = 0.01𝑁𝑚
while the equality of Froude number requires
1
𝑁𝑃 = 𝑁𝑚
√10
which shows two conflicting concepts.
If 𝜌𝑃 ≠ 𝜌𝑚 ; 𝜇𝑃 ≠ 𝜇𝑚 ;
𝜇 1 𝜇
[ ] = [ ]
𝜌 𝑚 31.6 𝜌 𝑃
Therefore, if kinematic viscosity of prototype is similar to water, the kinematic viscosity

of the fluid which needs to be employed for the model should be 1/31.6 of the kinematic
viscosity of water. It is impossible to find the fluid whose kinematic viscosity is that
small. As a conclusion, if all three dimensionless groups are important, it is impossible to
satisfy the dynamic similarity.
The specific oxygen demands and critical oxygen concentrations for typical microbial plants and
animal cell cultures are listed below
Cell culture qo Concentration, mmol/L
Escherichia Coli 0.5 mmol(gdw)-1h-1 0.0082
Vitisvinifera (grape) 0.6 mmol(gdw)-1h-1 0.055
Chinese Hamster Ovary 3.0x10-10 mmol(gdw)-1h-1 0.020
a.Estimate the kLa requirement to achieve cell concentrations of 25 gram dry weight/L for E. Coli
and V. Vinifera and 3.0x109 for CHO cells, while maintaining dissolved oxygen concentration
above critical. The oxygen solubility in the media used for the cultures is 7.2x10-3 kg/m3
𝑃𝑡
b. The relationship between kLa and the power input to a 1-m3 stirred bioreactor is kLa 𝛼 (𝑉𝑙 )0.5,
compare the bioreactor power requirements for culture of the three different cell types under the
conditions described in a.
Solution:
a.) for e coli: q= 8.5mmol/h (25/L)=292.65 mmol/L.h
v.vinifera: q= 0.6 mmol/h (25/L)=15 mmol/L.h
CHO: q= 3x10-10mmol/h (3x109/L)=0.9 mmol/L.h
Oxygen Solubilty: 7.3x10-3 kg/m3
292.5 mmol/L.h= [7.2 x10-3 kg/m3 (1kmol/32kg)(1x106 mmol/kmol)-0.0002)kla
kLa= 980.1661/hr
15 mmol/L.h= [7.2 x10-3 kg/m3 (1kmol/32kg)(1x106 mmol/kmol)-0.055)kla
kLa= 88.2353/hr
0.9 mmol/L.h= [7.2 x10-3 kg/m3 (1kmol/32kg)(1x106 mmol/kmol)-0.02)kla
kLa= 4.3902/hr
𝑃𝑡
b.) kLa= k(𝑉𝑙 )0.5
PT E.C.= 960725.5836 m3/hr2

K2
PT v.v. = 7765.4682 m3/hr2
K2
PT CHO= 19.2739 m3/hr2
K2
PT E.C.> PT v.v.> PT CHO
ESTIMATING KLa USING THE SIMPLE DYNAMIC METHOD
A stirred fermenter is used to haematopoietic cells isolated from umbilical cord blood. The liquid
volume is 15 Liters. The simple dynamic method is used to determine KLa. The air flow is shut off
for a few minutes and the dissolved oxygen level drops; the air supply is then reconnected at a
flow rate of 0.25 L/s. The following results are obtained at a stirrer speed of 50 rpm.
Time, s 5 20
Oxygen tension (%air saturation) 50 66
When steady state is established, the dissolved oxygen tension is 78% air saturation. In
separate test experiments, the electrode response to a step change in oxygen tension did not vary
with stirrer speed above 40 rpm. The probe response time under these conditions was 2.8 seconds.
When the KLa measurement was repeated using nitrogen to deoxygenate the mixture, the results
for oxygen tension as a function of time were similar to those listed. Estimate KLa.
Solution:
C∗ −C
ln(C∗AL− CAL1 )
AL AL2
KLa =
t2 − t1
78−50
ln(78−66)
KLa = = 0.056/s
(20−5)𝑠
STEADY-STATE KLa MEASUREMENTS
A 20-L stirred fermenter containing Bacillus thuringiensis is used to produce a microbial

insecticide. The oxygen balance method is applied to determine KLa. The fermenter operating
pressure is 150 KPa and the culture temperature is 30oC. The oxygen tension in the broth is
measured as 82% using a probe calibrated to 100% in situ using H2O and air at 30oC and 150KPa.
The solubility of oxygen in the culture fluid is the same as in H20. Air is sparged into the vessel;
the inlet gas flow rate measured outside the fermenter at 1 atm pressure and 22 oC is 0.23/s. The
exit gas from the fermenter contains 20.1% oxygen and has a flowrate of 8.91 min-1.
a. Calculate the volumetric rate of oxygen uptake by the culture
b. What is the value of KLa.
Solution:
1 𝐹𝑔 𝑃𝐴𝑔 𝐹𝑔 𝑃𝐴𝑔
a) NA = 𝑅𝑉 [( )𝑖 − ( ) 𝑜]
𝐿 𝑇 𝑇
0.23𝐿 1
1 ( 𝑠 ) (0.2099𝑎𝑡𝑚) (8.91𝑥 60) (0.201𝑥1.48
= [( )−( )]
0.08205𝐿. 𝑎𝑡𝑚 (22 + 273)𝐾 (30 + 273)𝐾
(20𝐿)
𝑚𝑜𝑙. 𝐾
1 𝑚𝑜𝑙
= (1.6357𝑥10−4 − 1.4572𝑥10−4 ) = 1.0878𝑥10−5
0.08205(20) 𝐿. 𝑠
Because of steady state the rate of oxygen transfer is equal to the rate of oxygen uptake
by the cells; the volumetric rate of oxygen uptake by the culture is 1.0878x10-5 mol/ L.s
b) Assume that the gas phase is well-mixed so that the oxygen concentration in the bubbles
containing the liquid is the same as the outlet gas, that is 20.1%. As the difference in the
composition of the gas phase to be constant throughout the fermenter.
The solubility of oxygen in H20 @30oC and 1atm air pressure is 8.05x10-3 kg/m3.
Determine the solubility at the fermenter operating pressure of 1.48 atm and gas phase
oxygen mole fraction of 0.201.
PT2 YAG1 1.48atm(0.201) 8.05x10−3 g

C*AL,2= C*AL,1 = ( )= 0.0114g/L
PT1 YAG2 1atm(0.2099) L
CAL in the fermenter is 82% of the oxygen solubility at 30oC and 1.48atm air pressure
1.48atm 8.05x10−3 g
CAL= 0.820( )( ) = 9.77x10-3 g/L
1atm L
32g
1.1x10−5 mol/L.s( )
mol
KLa = 0.0114g 9.77x10−3 g
KLa = 0.22 s-1
−
L L
DYNAMIC TECHNIQUE
A strain of Azobacter vinelandii is cultured in a 15m3 stirred fermenter for alginate production.
Under current operating conditions, KLa is 0.17 s-1. The solubility of oxygen in the broth is
approximately 8x10-3 kg/m3.
a. The specific rate of oxygen uptake is 12.5 mmol/g-hr. What is the maximum cell
concentration supported by oxygen transfer in the fermenter?
b. The bacteria suffer growth inhibition after copper sulphate is accidentally added to the
fermentation broth just after the start of the culture. This causes a reaction in the oxygen
uptake rate to 3 mmol/g-hr. What maximum cell concentration can now be supported
by oxygen transfer in the fermenter?
Solution:
8x10−3 kg
0.17
( s )( ) 𝟏𝟐𝟐𝟒𝟎𝐠
m3
a) Xmax = mmol 1hr 1mol 32g 1kg =
12.5 g.hr (3600s)(1000mol)(1mol)(1000g) 𝐦𝟑
b) Assume that addition of copper sulphate does not affect C*AL of KLa
8x10−3 kg
0.17
( s )( ) 𝟓𝟏𝟎𝟎𝟎𝐠
m3
Xmax= mmol 1hr 1mol 32g 1kg =
3 g.hr (3600s)(1000mol)(1mol)(1000g) 𝐦𝟑
GAS HANDLING WITH RUSHTON TURBINE
A fermenter of diameter and liquid height 1.4m is fitted with a Rushton impeller of diameter 0.5m
and off-bottom clearance 0.35m operated at 75 rpm. The fermentation broth is sparged with air at
a volumetric flow rate of 0.28m3/min. Half-way through the culture some bearings in the stirrer
drive begin to fail and stirrer speed must be reduced to a maximum of 45 rpm for the remainder of
the process.
a. Under the normal operating conditions, is the gas completely dispersed?
b. After the stirrer speed is reduced, is the impeller flooded or loaded?
Solution:
1min
a) Ni = 75/ min ( 60s ) = 1.25s-1
Ni2 Di (1.25 𝑠 −1 )2 (0.5𝑚)

Fr = = = 0.0796
g 9.81𝑚/𝑠 2
For complete gas dispersion Flg: Flooding- loading transition

D 0.5 0.5𝑚 0.5
Flg = 0.2 (D I ) Fr 0.5 = 0.2 (1.4𝑚) (0.0796)0.5 = 0.0337
T
Fg = FlgNiDi3 = (0.0337)(1.25s-1) (0.5m)3 = 5.27x10 -3 m3/s > 0.28 m3/min

*Fg, volumetric flowrate of gas greater than the operating flow rate, we can conclude that
the air provided is completely dispersed under normal conditions.
1min
b) Ni = 45/min( 60s ) = 0.75s-1
Ni2 Di (0.75 𝑠 −1 )2 (0.5𝑚)

Fr = = = 0.0287
g 9.81𝑚/𝑠 2
*Flooding-loading transition
D 3.5 0.5𝑚 3.5
Flg = 30 (D I ) Fr = 30 (1.4𝑚) = 0.0234
T
Fg = FlgNiDi3 = (0.0234)(0.75s-1) (0.5m)3 = 0.00219 m3/s

*At reduced stirrer speed, maximum air flow rate can be handled without impeller
flooding as operating flow rate (0.28m3/min) is greater than this. The impeller is
FLOODED.
Clostridium acetobutylicum carries out anaerobic fermentation and converts glucose into acetone,
butanol along with smaller concentrations of butyrate, acetate, etc. In fermentation the following
products were obtained from 100 moles of glucose and 11.2 moles of NH3, as nitrogen source.
Products formed:
Cells = 13moles acetic acid= 14 moles
Butanol= 56moles CO2= 221 moles
Acetone= 22moles H2= 135moles
Butyric acid = 0.4 moles Ethanol= 0.7 moles
By performing a carbon, nitrogen, hydrogen, and oxygen balance, determine the chemical
composition of the cells.
Solution:
By performing a carbon, hydrogen, nitrogen and oxygen balance, determine the element
composition of the cells.
100C6H12O6 + 11.2 NH3  13CaHbOcNd + 56C4H10O (butanol) + 22C3H6O(acetone) +
0.4C4H8O2(butyrate) + 14C2H14O2 (acetic acid) + 221CO2 +135H2 + 0.7 C2H6O(ethanol)
* where CaHbOcNd represents elemental composition of clostridium cells
Carbon Balance:
100(6) + 11.2(0) = 13(a) + 56(4) + 22(3) + 0.4(4) + 14(2) + 221(1) + 0.7(2) : a = 4.46
Hydrogen Balance:
100(12) + 11.2(3) = 13(b) + 56(10) + 22(6) + 0.4(8) + 14(14) + 135(2) + 0.7(6) : b = 16.02
Oxygen Balance:
100(6) + 11.2(0) = 13(c) + 56(1) + 22(1) + 0.4(2) + 14(2) + 221(2) + 0.7(1) : c = 3.88
Nitrogen Balance:
11.2(1) = 13(d) : d = 0.86
Chemical Composition of the cell = C4.46H16.02O3.88N0.86

Aerobic Culture of Saccharomyces cerevisae in a synthetic medium produced the following:

µ= 0.2/hr
RQ =1.0
Yy/s = 90 grams dry cell/gmol glucose
Vglutamate = 0.043 grams glutamate/ g cell-hr
Assuming that carbon content of the cell is 45%, check carbon and oxygen balance with
respect to given culture.
aC6H12O6 + bC5H9O4N + cO2  dCHαOβOδ + e CO2 + fH2O

glutamic acid
Solution:
RQ = 1.0 = Q02 = QCO2 = c = e
@ µ = 0.2 hr -1 and 45% C in biomass ; 1mol cell biomass = 12gC
0.2
(0.45)
hr
d= = 7.5x 10-3 mol C/g dry cell. Hr
12g/mol
@100 g cell biomass containing 45g C (45% C)
g cell biomass
C
g Carbon
45% = (100)
MW cell biomass
12(100)
MWcell = 45 = 26.67
MW = 26.67 g/mol = CHαOβNδ
26.67= 1(12) + α(1) + β(16) + δ(14)
14.67= α + 16β + 14δ (Equation 1)
µ 0.2/hr
a= Qglucose = = g dry cell = 2.22x10-3 gmol glucose/ g dry cell. Hr
YX/S 90
g glucose
g glutamate
0.043
g cell.hr
b= Qglutamate = g = 2.925x10-4 gmol glutamate/ g dry cell. Hr
147 glutamic acid
mol
Elemental Balance:
C: 6a + 5b = d + e
H: 12a + 9b = dα + 2f
O: 6a + 4b + 2c = βd + 2e + f : 2c=2e : 6a + 4b = βd + f
N: b = δd
b 2.925 𝑥 10−4
δ= = = 0.0390
d 7.5𝑥 10−3
Using H and O:
12a + 9b = dα + 2f
-2 (6a + 4b = βd + f)
b= αd - 2βd since b = 2.925x10-4 : d = 7.5x10-3
2.925x10-4 = α (7.5x10-3) - 2β (7.5x10-3)

2.925𝑥10−4 + 0.015𝛽
α=
7.5𝑥10−3
from equation 1:
14.67 = α + 16β + 14δ : 14.67 = α +16β + 14(0.0390)
14.124 = α + 16β
Substituting α:
2.925𝑥10−4 + 0.015𝛽
14.124 = + 16β : β = 0.7825, α = 1.604
7.5𝑥 10−3
Using equation of O:
6a + 4b = βd + f ; f = 6a + 4b – βd
f= 6(2.22x10-3) + 4(2.925x10-4) – 0.7825(7.5x10-3)
f = 8.6213x10-3
Using equation of C:
6a + 5b = d + e ; e = 6a + 5b – d
e = 6( 2.22x10-3) + 5( 2.925x10-4) – 7.5x10-3
e = c = 7.2825x10-3
* aC6H12O6 + bC5H9O4N + cO2  dCHαOβOδ + e CO2 + fH2O
Answer:
2.22 C6H12O6 + 2.925C5H9O4N + 7.2825 O27.5 CH1.604O0.7825O0.0390 + 7.2828 CO + 8.6213H2O
A 50 m3 bioreactor (H/DT=2.5; working volume=60%) equipped with two sets of a standard flat
blade turbine is used for yeast growth, the bioreactor is operated continuously at a dilution rate of
0.3 hr-1. The organism obeys the Monod’s equation (μm=0.4 hr-1 and Ks=2 kg/m3). The inlet sugar
feed concentration is 50 kg/m3. The bioreactor is aerated and agitated at 0.5 vvm at 60 rpm. The
yield of biomass based on glucose is 0.5 g cell (dry) per gram glucose consumed. The density and
viscosity of the broth are 1200 kg/m3 and 0.02 Pa•s.
Cell formula: CH1.8O0.5N0.2
State whether the system is mass transfer limited or biochemical reaction limited.
Given:
VT=50 m3 D=0.3 hr-1
H/DT=2.5 μm=0.4 hr-1
working volume=60% Ks=2 kg/m3
two sets of a standard flat blade turbine; Pmo=2 Pmo CSO=50 kg/m3
Q=0.5 vvm 0.5 g cell (dry)
YX/S= 𝑔 𝑔𝑙𝑢𝑐𝑜𝑠𝑒
N=60 rpm
ρ = 1200 kg/m3
μ=0.02 Pa•s
Cell formula: CH1.8O0.5N0.2
Required: Mass transfer limited or Biochemical reaction limited

Sol’n:
𝐻
 = 2.5
𝐷𝑇
H = liquid height
DT = tank diameter
𝜋 𝜋 𝜋
 𝑉𝑇 = 𝐷𝑇 2 𝐻 = 𝐷𝑇 2 (2.5𝐷𝑇 ) = (2.5) (𝐷𝑇 3 )
4 4 4
VT = 50 m3
𝜋 50 𝑚3
50 𝑚3 = (2.5) (𝐷𝑇 3 ) ; 𝐷𝑇 = 3√ 𝜋
4 (2.5)
4
DT = 2.9420 m
𝜇 0.02 𝑃𝑎∙𝑠
 𝜈= = 1200 𝑘𝑔/𝑚3 = 1.67 × 10−5 𝑚3 /𝑠
𝜌
𝐷𝑇
 From Biochemical Engineering by James Lee: = 3 ; DI = impeller diameter
𝐷I
𝐷𝑇 2.9420 𝑚
𝐷I = = = 0.9807 𝑚
3 3
 For an air-electrolyte solution:

𝑃𝑚 0.70
𝑘𝐿 𝑎 = 2.0 × 10−3 [ ] 𝑉𝑠 0.2 (eq. 9.72, p. 267, James Lee)
𝑉𝐿
kLa =volumetric mass- transfer coefficient

Pm = gassed power
VL = volume of the liquid or broth
Vs = superficial velocity
 For the power number, Np,

1 𝑚𝑖𝑛 2 𝑘𝑔
𝑁𝐷 2
𝜌 60𝑟𝑝𝑚 ( ) (0.9807 𝑚) (1200 )
𝑅𝑒𝑦𝑛𝑜𝑙𝑑 ′ 𝑠 𝑁𝑢𝑚𝑏𝑒𝑟, 𝑅𝑒 =
𝐼
=
60 𝑠𝑒𝑐 𝑚3
𝜇 0.02 𝑃𝑎 ∙ 𝑠
= 57706.3494
 Re ≥ 10000; Np = 6 (p. 258, James Lee)
N = (60/60) rps
 For a flat-blade turbine at Re≥ 10000

𝑘𝑔 60
𝑃𝑚𝑜 = 𝑁𝑃 𝜌 𝑁 3 𝐷𝐼 5 = 6 (1200 𝑚3 ) (60 𝑟𝑝𝑠)3 (0.9807 𝑚)5
𝑃𝑚𝑜 = 6531.5066 𝑊 (𝑢𝑛𝑔𝑎𝑠𝑠𝑒𝑑 𝑝𝑜𝑤𝑒𝑟)
 For the gassed power

𝐷
0.115 1.96 ( 𝐼 )
𝑃𝑚 𝐷𝐼 4.38 𝐷𝐼 2 𝑁 𝐷𝐼 𝑁 2 𝐷𝑇 𝑄
log10 𝑃 = −192 [𝐷 ] [ ] [ ] [𝑁 𝐷 3 ] (eq. 9.53, p. 258,
𝑚𝑜 𝑇 𝜈 𝑔 𝐼
James Lee)
0.5 𝑚3 𝑂2
𝑄= (50 𝑚3 × 0.60) = 0.25 𝑚3 /𝑠
𝑚3𝑏𝑟𝑜𝑡ℎ∙𝑚𝑖𝑛
𝑃𝑚
log10 2 (6531.5066 =
𝑊)
0.9807
2 60
0.115 60 1.96 ( )
0.9807 𝑚 4.38 (0.9807 ) (60 𝑟𝑝𝑠) (0.9807 ) ( )2 2.9420
0.25 𝑚3 /𝑠
60
−192 [2.9420 𝑚] [ 1.67 ×10−5
] [ 9.81 𝑚/𝑠2
] [ 60 ]
( ) (0.9807 )3
60
Pm = 6191. 1993 W
 For computation of kLa, superficial velocity
1 𝑚𝑖𝑛
0.5 𝑚3 𝑎𝑖𝑟 ( )
60 𝑠𝑒𝑐
𝑄 𝑚3 𝑚𝑒𝑑𝑖𝑎 ∙𝑚𝑖𝑛
𝑉𝑠 = = 𝜋 (50 𝑚3 × 0.60) = 0.0368 𝑚/𝑠
𝑆 (2.9420 𝑚)2
4
6191.1993 𝑊 0.70 𝑚
𝑘𝐿 𝑎 = 2.0 × 10−3 [ 50 𝑚3 (0.60) ] (0.0368 𝑠 )0.20 = 0.0431 𝑠 −1
 Assume: Solution of H2SO4 @ 2.0 mols/L, find the corresponding oxygen solubility from
Table 9.2 of Biochemical Engineering by James Lee, p. 262
CO2 = 1.02 mmol/L
 Use the solubility of O2 from Table 9.2 to determine the Henry’s constant and the
equilibrium concentration of O2 at that condition.
1 𝑎𝑡𝑚 𝑎𝑡𝑚 ∙𝐿
𝐻𝑂2 = 1.02 𝑚𝑚𝑜𝑙/𝐿 = 0.9804 𝑚𝑚𝑜𝑙
0.21 𝑎𝑡𝑚 𝑚𝑚𝑜𝑙
𝐶𝐿 ∗ = 𝑎𝑡𝑚 ∙𝐿 = 0.2142
0.9804 𝐿
𝑚𝑚𝑜𝑙
 For kLa,CL*, the volumetric mass-transfer coefficient at the equilibrium:
0.0431 3600 𝑠 𝑚𝑚𝑜𝑙 1 𝑚𝑜𝑙 1 𝑘𝑚𝑜𝑙 32 𝑘𝑔 1000 𝐿
𝑘𝐿 𝑎,𝐶𝐿∗ = 𝑠 [ 1 ℎ𝑟 ] (0.2142 𝐿 ) [1000 𝑚𝑚𝑜𝑙] [1000 𝑚𝑜𝑙] [1 𝑘𝑚𝑜𝑙] [ 1 𝑚3 ] =
𝑘𝑔
1.0635 𝑚3 ∙ ℎ𝑟
 Biochemical reaction limited: Using Monod Equation

𝐶
𝐷 = 𝜇 = 𝜇𝑚 (𝐾 +𝑠 𝐶 )
𝑠 𝑠
 Determine the Exit substrate concentration, Cs
0.3 0.4 𝐶𝑠
= [ 𝑘𝑔 ] ; 𝐶𝑠 = 6.0 𝑘𝑔/𝑚3
ℎ𝑟 ℎ𝑟 2 3 + 𝐶𝑠
𝑚
 Then, determine the cell concentration that was produced

𝑔 𝑑𝑟𝑦 𝑐𝑒𝑙𝑙 𝑘𝑔 𝑘𝑔
𝐶𝑥 − 𝐶𝑥𝑜 = 𝑥 = 𝛾𝑥 ( 𝐶𝑠𝑜 − 𝐶𝑠 ) = 0.5 𝑔 𝑔𝑙𝑢𝑐𝑜𝑠𝑒 𝑐𝑜𝑛𝑠𝑢𝑚𝑒𝑑 (50 − 6 ), 𝑚3 = 22 𝑚3
𝑠
 For ZO2, grams of oxygen required per gram dry cell produced (Maleles, 1971)
32𝐶+8𝐻−16𝐶
𝑍𝑂2 = 𝛾 ∙ 𝑀𝑊 + 0.01𝑂′ − 0.0276𝐶 ′ + 0.01714𝑁 ′ − 0.08𝐻 ′
𝑥
𝑠
Where:
C, H, O – is the number of atoms present in the carbon source
C’, H’, O’, N’, - is the percent carbon, hydrogen, oxygen and nitrogen respectively in the
cell
*From C6H12O6 (carbon source) *From CH1.8O0.5N0.2 (cell formula)
C= 6 C= 1(12) C’= (1(12)/ 24.6) x100 =

48.78
H=12 H= 1.8(1) H’= (1.8(1)/ 24.6) x100 =
7.32
O=6 O= 0.5(16) O’= (0.5(16)/ 24.6) x100 =

32.52
MW= 6(12) + 12(1) + 6(16) N= 0.2(14) N’= (0.2(14)/ 24.6) x100 =

11.38
∑= 24.6
32(6)+8(12)−16(16)
𝑍𝑂2 = + 0.01(32.52) − 0.0276(48.78) + 0.01714(11.38) −
0.5 (180)
0.08(7.32)
𝑔 𝑂2
𝑍𝑂2 = 0.7656 𝑔 𝑑𝑟𝑦 𝑐𝑒𝑙𝑙
𝑔𝑂 𝑘𝑔 0.4 𝑘𝑔
𝑍𝑂2 ∙ 𝑥 ∙ 𝜇𝑚 = 0.7658 𝑔 𝑑𝑟𝑦 2𝑐𝑒𝑙𝑙 (22 𝑚3 ) ( ℎ𝑟 ) = 6.73904 𝑚3 ∙ℎ𝑟
ZO2(x)(μm) > kLa, Cl*

Biochemical reaction > Mass Transfer
Thus, it is Mass transfer limited because there’s very little oxygen that is being
transferred or dissolved in the broth.
In an oxygen absorption study, the following data were obtained on the oxygen transfer capacity
of an air diffusion unit.
Q= 9.439 m3/s Air bubble diameter: 2.5 x 10-3 m
T= 12°C Air bubble velocity: 0.3 m/s
HL= 4.27 m DT= 99.5 m
The dissolved oxygen concentration measurement was tabulated as:

Time, min Cs, kg/m3
3 0.6 x10-3
6 1.6 x10-3
9 3.1 x10-3
12 4.3 x10-3
15 5.4 x10-3
18 6.0 x10-3
21 7.0 x10-3
From the information, compute the:

a. KLa and kL
b. Mass of oxygen per hour transferred per 28.317 m3 and zero dissolved oxygen
concentration and the oxygen transfer efficiency
c. How much oxygen will be transferred to waste with α=0.80 at temperature of 32°C and an
operating dissolved oxygen of 15 x10-3 kg/m3? Assume the saturation concentration of
oxygen in the liquid at 12°C to be 1.08 x10-2 kg/m3, at 20°C to be 1.07 x10-2 kg/m3 and at
32°C to be 8.673 x10-2 kg/m3
Given:
Cs= 1.08 x10-2 kg/m3
Solution:
𝑃 𝑂
A. 𝐶𝑠𝑚 = 𝐶𝑠 [29.4𝑏
+ 42𝑡 ]
Pb - absolute pressure at the depth of air release, psi
Ot – concentration of O2 in air leaving the tank, %
Cs - saturation concentration of O2 @ experimental temperature
Csm – mean oxygen saturation concentration
 @ 10% absorption
𝑚𝑜𝑙𝑒𝑠 𝑜𝑓 𝑢𝑛𝑎𝑏𝑠𝑜𝑟𝑏𝑒𝑑 𝑂2 21 (1−0.1)
𝑂𝑡 = 𝑡𝑜𝑡𝑎𝑙 𝑚𝑜𝑙𝑒𝑠 𝑜𝑓 𝑙𝑒𝑎𝑣𝑖𝑛𝑔 𝑎𝑖𝑟 × 100 = × 100 = 19.3 %
21 (1−0.1)+79
 For Csm,
𝑘𝑔 20.2 𝑝𝑠𝑖 19.3 𝑘𝑔
𝐶𝑠𝑚 = 1.08 x 10−2 𝑚3 [ 29.4 + ] = 0.01238 𝑚3
42
Time, min Csm – Cs, kg/m3
3 0.01238 - 0.6 x10-3 0.01178
6 0.01238 - 1.6 x10-3 0.01078
9 0.01238 - 3.1 x10-3 9.28 x10-3
12 0.01238 - 4.3 x10-3 8.08 x10-3
15 0.01238 - 5.4 x10-3 6.98 x10-3
18 0.01238 - 6.0 x10-3 6.38 x10-3
21 0.01238 - 7.0 x10-3 5.38 x10-3
 Plot Csm – Cs vs. Time

𝑘𝑔 60 𝑚𝑖𝑛 𝑘𝑔
𝑚 = 𝑠𝑙𝑜𝑝𝑒 = −3.6071 × 10−4 𝑚3 ∙𝑚𝑖𝑛 [ 1 ℎ𝑟 ] = −0.0216 𝑚3 ∙ℎ𝑟
 Determine the kLa
𝑘𝑔
𝑠𝑙𝑜𝑝𝑒 −0.0216 3
𝑘𝐿 𝑎 = 𝑚𝑒𝑎𝑛 ∆ 𝑐𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 𝑜𝑓 𝑂 = 𝑚 ∙ℎ𝑟
𝑘𝑔 = 2.7 ℎ𝑟 −1
2 (0−8×10−3 ) 3
𝑚
 Determine the interfacial area for the calculation of the mass transfer coefficient, kL
𝐴 6𝑄𝐻 𝜋
= 𝑑 𝑉 𝑉𝐿 ; 𝑉𝑇 = 4 𝐷𝑇 2 𝐻𝐿
𝑉 𝐵 𝐵 𝑇
Where :
A/V – is the interfacial area
dB – is the air bubble diameter
VB - is the air bubble velocity
𝜋
𝑉𝑇 = (99.5 𝑚)2 (4.27𝑚) = 33,201.97 𝑚3
4
𝐴 6 (9.439 𝑚3 )(4.27 𝑚)
= 𝑚 = 9.7114 𝑚−1
𝑉 (2.5 ×10−3 𝑚)(0.3 )(33,201.97 𝑚3 )
𝑠
 For kL,
𝑘𝐿 𝑎 2.7/ℎ𝑟 𝑚
𝑘𝐿 = 𝐴/𝑉 = = 0.2677 ℎ𝑟
9.7114/𝑚
B. 𝑘𝐿 𝑎(𝑇) = 𝑘𝐿 𝑎(20℃) (1.02)𝑇−20

𝑘𝐿 𝑎(12℃) 2.6/ℎ𝑟 3.046
𝑘𝐿 𝑎(20℃) = 1.0212−20
= 1.0212−20
= ℎ𝑟
𝑞
 (𝑣 ) = 𝑘𝐿 𝑎(20℃) (𝐶𝑠,20℃ − 𝐶𝐿,20℃ ) (eq.1)
(20℃)
𝑞
 (𝑣 ) = 𝑘𝐿 𝑎(32℃) (𝐶𝑠,32℃ − 𝐶𝐿,32℃ ) (eq.2)
(32℃)
𝑘𝐿 𝑎(32℃) = 𝑘𝐿 𝑎(20℃) (1.02)𝑇−20
 Determine the amount of oxygen transferred per hour at 20°C.

𝑞 3.046 𝑘𝑔
(𝑣 ) = 𝑘𝐿 𝑎(20℃) (𝐶𝑠 ∗ − 𝐶𝐿,20℃ ) = ℎ𝑟 (1.07 × 10−2 − 0) 𝑚3
(20℃)
3.046 𝑘𝑔 𝑘𝑔
𝑞(20℃) = (1.07 × 10−2 − 0) (28.317 𝑚3 ) = 0.923
ℎ𝑟 𝑚3 ℎ𝑟
C. From eq.1 and 2, in terms of equal working volume,

𝑘𝐿 𝑎(32℃) (𝐶𝑠,32℃ −𝐶𝐿,32℃ )
𝑞(32℃) = 𝑘 𝑞
𝐿 𝑎(20℃) (𝐶𝑠,20℃ −𝐶𝐿,20℃ ) (20℃)
𝑘𝐿 𝑎(20℃) (1.02)32−20 (𝐶𝑠,32℃ −𝐶𝐿,32℃ )
𝑞(32℃)@ 80% 𝑒𝑓𝑓 = 0.80 [ ] 𝑞(20℃)
𝑘𝐿 𝑎(20℃) (𝐶𝑠,20℃ −𝐶𝐿,20℃ )
𝑘𝑔 𝑘𝑔
(0.923 )(8.673×10−2 −1.5×10−2 ) 3 (1.02)32−20 𝑘𝑔
ℎ 𝑚
𝑞(32℃)@ 80% 𝑒𝑓𝑓 = 0.80 [ ] = 6.2778 ℎ𝑟
(1.07×10−2 −0)
The usual procedure of scale up of fermenter is to fix one of several criteria involving Reynolds
Number, power consumption per unit volume of liquid, tip velocity of an impeller, the liquid
circulation time and the volumetric oxygen transfer coefficient. The choice of criterion will
depend on the fermentation being studied. Estimate, using two methods, the required speed of an
impeller and the power requirements of a production scale fermenter of 60 m3, to match the
volumetric mass transfer coefficient. Following optimum conditions were obtained with a 0.03 m3
fermenter:
Density of broth: 1200 kg/m3 Liquid volume: 0.018 m3
Aeration rate: 1vvm Oxygen transfer rate: 0.25 kmol/m3•hr
Liquid Height: 1.2 DT
Two sets of standard flat blade turbine impellers were installed.

Given:
ρL= 1200 kg/m3 HL= 1.2 DT VL= 0.018 m3
Q= 1.0 vvm OTR= 0.25 kmol/m3•hr
V1= 0.03 m3 V2= 60 m3
Solution:
 Determine the tank diameter, DT, and impeller diameter, DI, for each conditions.
𝜋 𝜋
𝑉1 = 𝐷𝑇1 2 𝐻𝐿 = (𝐷𝑇1 )2 (1.2 𝐷𝑇1 ) = 0.3𝜋𝐷𝑇1 3
4 4
1 1
𝑉1 3 0.018𝑚3 3
𝐷𝑇1 = (0.3𝜋) = [ ] = 0.267 𝑚
0.3𝜋
1 1
𝐷𝐼1 = 3 𝐷𝑇1 = 3 (0.267 𝑚) = 0.089 𝑚 (Biochemical Engineering by James Lee,
pg.274)
𝐻𝐿1 = 1.2 𝐷𝑇1 = 1.2 (0.267 𝑚) = 0.32 𝑚
1
1 0.018 3
𝑉2 3 60 𝑚3 ×
0.03
𝐷𝑇2 = (0.3𝜋) = [ ] = 3.36 𝑚
0.3𝜋
1 1
𝐷𝐼2 = 3 𝐷𝑇2 = 3 (3.36 𝑚) = 1.12 𝑚
𝐻𝐿2 = 1.2 𝐷𝑇2 = 1.2 (3.36 𝑚) = 4.03 𝑚
 Aeration flow rate:

1 𝑚3 𝑂 1𝑚𝑖𝑛
2
𝑄 = 𝑚3 𝑏𝑟𝑜𝑡ℎ ∙𝑚𝑖𝑛 (0.018 𝑚3 ) [ ] = 3 × 10−4 𝑚3 /𝑠𝑒𝑐
60𝑠𝑒𝑐
 For superficial velocity

𝑚3 3600𝑠𝑒𝑐
𝑄 3×10−4 [ ] 𝑚
𝑠𝑒𝑐 1ℎ𝑟
𝑉𝑠 = 𝐴 = 𝜋 2
= 19.3 ℎ𝑟
(0.267𝑚)
4
 Determine the partial pressure of oxygen in the system

𝐻𝐿 0.32 𝑚 𝐻2 𝑂
1 𝑎𝑡𝑚+(1 𝑎𝑡𝑚+ ) 1 𝑎𝑡𝑚+[1𝑎𝑡𝑚+ ]
10.3𝑚 𝐻2 𝑂 10.3 𝑚 𝐻2 𝑂
𝑃𝑂2 = (0.21) = (0.21) = 0.213 𝑎𝑡𝑚
2 𝑎𝑡𝑚 2 𝑎𝑡𝑚
 Then determine the kLa

𝑘𝑚𝑜𝑙
𝑂𝑇𝑅 0.25 3 𝑘𝑚𝑜𝑙
𝑚 ∙ℎ𝑟
𝑘𝐿 𝑎 = = = 1.17
𝑃 𝑂2 0.213 𝑎𝑡𝑚 𝑚3 ∙ℎ𝑟∙𝑎𝑡𝑚
 For the determination of the power requirement

 Cooper et. al. (1944) correlated kLa, gassed power per unit volume (Pm/V)and superficial
velocity (Vs) for Varied Disk Impellers as:
𝑃 0.95 𝑘𝑚𝑜𝑙
𝑘𝐿 𝑎 = 0.0635 ( 𝑉𝑚 ) (𝑉𝑠 )0.67 , 𝑚3 ∙ℎ𝑟∙𝑎𝑡𝑚
 For Standard-Flat Blade Turbine, Aiba et.al. (1965)
𝑃 0.95 𝑘𝑚𝑜𝑙
𝑘𝐿 𝑎 = 0.0318 ( 𝑉𝑚 ) (𝑉𝑠 )0.67 , 𝑚3 ∙ℎ𝑟∙𝑎𝑡𝑚
 For Turbine Type Impellers, Van’t Riet (1979)
𝑃 0.4 𝑘𝑔
𝑘𝐿 𝑎 = 0.026 ( 𝑉𝑚 ) (𝑉𝑠 )0.5 , 𝑚3 ∙ℎ𝑟
 For the gassed power,Pm

𝑘𝑚𝑜𝑙 𝑃 0.95 𝑚
1.17 𝑚3 ∙ℎ𝑟∙𝑎𝑡𝑚 = 0.0318 (0.018𝑚𝑚3 ) (19.3 ℎ𝑟)0.67 = 0.1 ℎ𝑃
 Then determine the ungassed power, Pmo, for two sets of impeller, Np= 2x6=12
𝑃 𝑔 𝑁𝑝 𝜌 𝑁𝑖 3 𝐷𝑖 5
𝑁𝑝 = 𝜌 𝑁𝑚𝑜3 𝐷𝑐 5 ; 𝑃𝑚𝑜 =
𝑖 𝑖 𝑔𝑐
𝑘𝑔
𝑁𝑝 𝜌 𝑁𝑖 𝐷𝑖 3 5 12 (1200 3 ) (𝑁𝑖 )3 (0.089 𝑚)5 𝑘𝑔∙𝑚
𝑃𝑚𝑜 = = 𝑚
𝑚 = 8.197 × 10−3 𝑁𝑖 3 , ≈ 0.108 ×
𝑔𝑐 9.81 2 𝑠
𝑠
10−3 𝑁𝑖 3
 Using Michaelis correlation:

2
(𝑃𝑚𝑜1 ) 𝐷𝑖1 3
𝑃𝑚1 = 0.5 [ ]
𝑄1 0.56
2
(0.108 ×10−3 𝑁𝑖 3 ) (0.089 𝑚)3
0.1ℎ𝑃 = 0.5 [ ]
(3×10−4 𝑚3 /𝑠𝑒𝑐)0.56
60𝑠𝑒𝑐
𝑁𝑖 = 18 𝑟𝑝𝑠 [ 1𝑚𝑖𝑛 ] = 1080 𝑟𝑝𝑚
𝑃𝑚𝑜1 = 0.108 × 10−3 𝑁𝑖 3 , ℎ𝑃 = 0.108 × 10−3 (1080)3 = 0.630 ℎ𝑃
𝑃 0.10
[𝑃 𝑚 ] = 0.630 = 0.1587
𝑚𝑜 1
 For Constant Power Input in a Baffled Vessel

𝑃𝑚𝑜 𝑃
= 𝜌∙𝑁 𝑚𝑜
3 ∙𝐷 5 ; 𝑤ℎ𝑒𝑟𝑒 𝑃𝑚𝑜 = 𝑐𝑜𝑛𝑠𝑡𝑎𝑛𝑡 𝑎𝑛𝑑 𝜌 = 𝑀/𝑉
𝜌∙𝑁 3 ∙𝐷 5
1 𝐼1 2 𝐼2
1 1
[𝑀 3 ∙𝐷 5
] = [𝑀 ]
∙𝑁1 𝐼1 ∙𝑁2 3 ∙𝐷𝐼2 5
𝑉1 𝑚𝑜𝑑𝑒𝑙 𝑉2 𝑝𝑟𝑜𝑡𝑜𝑡𝑦𝑝𝑒
 For constant power in geometrically similar vessels

𝜌∙𝑁1 3 ∙𝐷𝐼1 5 𝜌∙𝑁2 3 ∙𝐷𝐼2 5
=
𝑉1 𝑉2
*Accdg. To Ghose *Accdg to James Lee; Pm/V= constant

1 5
2 2/3
𝑁1 3 ∙𝐷𝐼1 5 𝑉2 3 𝐷𝐼 3 𝑁 3 𝐷𝐼2 𝐷𝐼2
𝑁2 3 = 𝑉1 ; 𝑁2 = 𝑁1 [𝑉 ] [𝐷 1 ] [ 1] = [ ] ; 𝑁2 = 𝑁1 [ ]
𝐷 5 1 𝐼2 𝑁2 𝐷𝐼1 𝐷𝐼1
𝑉2 𝐼2
−2/3
1 5 𝐷𝐼 0.089 −2/3
𝑁2 = 1080
60 3 0.089 3
[0.03] [ 1.12 ] , 𝑟𝑝𝑚 𝑁2 = 𝑁1 [𝐷 2 ] = 720 [ 1.12 ]
𝐼1
𝑁2 = 199.8575 𝑟𝑝𝑚 𝑁2 = 3895.39 𝑟𝑝𝑚
*for constant impeller tip velocity *for constant impeller speed

𝑁1 𝐷𝐼1 = 𝑁2 𝐷𝐼2 2/3
𝐷𝐼
𝑁1 = 𝑁2 [𝐷 2 ] ; 𝑁𝑝 =
𝐷𝐼1 0.089 𝐼1
𝑁2 = 𝑁1 [𝐷 ] = 720 [ 1.12 ] 2/3
𝐼2 𝐷𝐼
𝑁2 = 57.2143 𝑟𝑝𝑚 𝑁𝑚 [ 𝐷 𝑚 ]
𝐼𝑝
𝑁2 = 3895.39 𝑟𝑝𝑚
Or
(𝑁𝐷𝐼 )𝑝 = 1.7 (𝑁𝐷𝐼 )𝑚
CHAPTER 9: AGITATION AND AERATION ; ADDITIONAL PROBLEM
A strain of azobacter vinelandii is cultured in a 15 m3 stirred fermenter for production of alginate.

Under conditions the mass transfer coefficient, KLa, is 0.25 s-1. Oxygen solubility in a fermenter
10−3 𝑘𝑔
broth is approximately 8.5 × . The specific oxygen uptake is 15 mmol g/h. What is the max
𝑚3
cell density in the broth?

REQ’D: Xmax
Sol’n:
KLa×𝐶 ∗ 𝑎𝑙
Xmax = 𝑞𝑜2
0.25 𝑘𝑔
( )(8.5∗10−3 3 )
𝑠 𝑚
= 𝑚𝑚𝑜𝑙 1 1 32𝑔 1𝑘𝑔

(15 )( )( )( )( )
𝑔∗ℎ 3600𝑠 1000𝑚𝑚𝑜𝑙 1𝑚𝑜𝑙 1000𝑔
𝑔 𝑔
Xmax = 15937.5 𝑚3 = 15.94 𝐿
Biochemical Engineering and Biotechnology by Ghasem D. Najafpour
Calculate the gas hold-up for an agitated and aerated system with power input of 18 hp in an 80
m3 vessel with gas superficial velocity of 2.6 m.min-1
Given:
P = 18 hp
V = 80 m3
Vs = 2.6 m/min
Required:
Gas Hold-up, Ho
Solution:
From equation;
𝑃
(𝑉)0.4 (𝑉𝑠)0.5 = 7036𝐻 + 2.37
Where; P = power in hp
V = ungassed liquid volume in m3
Vs = gas superficial velocity in m/h
Solving for H:
18
(80)0.4 (2.6 𝑥 60 𝑚𝑖𝑛/ℎ)0.5 = 7.36𝐻 + 2.37
H = 0.6 m
The gas hold up can be defined by the above definition using the gas height per volume,
where H = 0.6 m for aeration
Vg 0.6
Ho = Vg+VL = 0.6+6.5 = 0.085
Ho = 8.5%
Biochemical Engineering and Biotechnology by Ghasem D. Najafpour
Calculate the speed of an impeller and the power requirements of a production-scale bioreactor
with 60m3 using two different methods. Also, match the volumetric mass transfer coefficient. The
following optimum conditions were obtained with a small scale fermenter of volume 0.3m3 and
60% of the vessel working. The density of the broth, ρbroth, 1200 kg.m-3, working volume 0.18m3,
aeration rate of one volume of gas per volume of liquid (vvm), oxygen transfer rate 0.25 kmol.m -
3
.h-1, liquid height inside the vessel, HL, 1.2Dt. two sets of standard, flat-blade turbine impellers
were installed.
Given:
@ production-scale, V2 = 60m3
@ small-scale, V = 0.3m3
60% of vessel working
ρbroth, 1200 kg.m-3
working volume, V1 = 0.18m3
Q = 1vvm
OTR = 0.25 kmol.m-3.h-1
HL = 1.2Dt
Two sets of standard flat-blade turbine impellers
Required: impeller speed, power requirement, kla
Solution:
V1 = (π/4)(Dt)2(1.2Dt) = 0.3πDt3
Diameter of the vessel:
Dt = (V1/0.3π)1/3 = (0.18/3π)1/3 = 0.576m
Diameter of the impeller:
Di,1 = (1/3)(Dt) = 0.576/3 = 0.192m
Height of liquid media was assumed to be 1.2 times the diameter of the fermenter vessel.
HL,1 = 1.2 x 0.576 = 0.691m

Diameter of the larger vessel:
Dt2 = (V1/0.3π)1/3 = (60/0.3π)1/3 = 3.36m
The impeller size for the larger vessel is:
Di2 = 3.36/3 = 1.12m
And the liquid media height in the second fermenter is:
HL2 = 1.2 x 3.36 = 4.03m
Assume the fermentation broth has the same viscosity as water:
µ1 = 1cp
Aeration rate for 1vvm is:

𝜋
Us = (0.18 x 60)/( 4 x 0.5762) = 41.45 m/h
Let us take average values for the partial pressure of oxygen
PO2 = ({1 atm + [1 + (HL m/10.3m) x atm]}/2) x 0.21 = 0.213 atm
The oxygen transfer rate is
OTR = 0.25 kmol/m3.h
The mass transfer coefficient is
kL = 0.25/0.213 = 1.174 kmol/m3.h.atm
Use imperial correlation based on the following equation for mass transfer in the bioreactors.
The general equation for the evaluation of Kla is
Kla = x(Pg/V)Y (Us)Z
where x,y, and z are empirical constants. For Newtonian fluids, mnon-coalescing broth and gas
bubbles, the following correlation is valid for a working volumeof less than 4 m3 and a power per
unit volume of 500 – 10,000 W/m3
kL = 0.002(Pg/V)0.7(Us)0.5
1.174 = 0.002 (Pg/V)0.7(41.45)0.5
For the gassed power per unit volume (Pg/V) is 630.7 W; thegpassed power, Pg, was 0.15hp.
Since the flow regime is turbulent, the power number obtained from figure 6.6, Power number
versus Reynolds number, Re, reads Pno = 6. For two sets of impellers, Np = 2(6) = 12
` Np = (P1 gc)/(ρN13Di5)
P1 = (12 x 1200 x N13 x 0.1925)/9.81 = 0.383 N13 W = 5x10-4 N13 hp
Using Michel and Miller’s correction factor for power calculation:
Pg1 = 0.5(P12η1Di13/µ10.56)0.45
Knowing the power input, we can calculate the rotational speed:
0.15 = 0.5(5x10-4 x N13 x 0.1923 / (3x10-3)0.56)0.45
N1 = 10 rps
N1 = 600 rpm
The power input for ungassed system is
P1 = 5x10-4 N13 = 5x10-4 (10)3 = 0.5hp
Pg/P = 0.15/0.5 = 0.3
For constant power input based on geometric similarity of the vessels, agitation rate is calculated.
(ρN13Di15)/(V1) = (ρN23Di25)/(V2)
(N2/N1)3 = (V2/V1)(Di1/Di2)5
N2 = N1 (V2/V1)1/3(Di1/Di2)5/3 = 600(60/0.3)1/3(0.192/1.12)5/3 = 185 rpm
For constant input velocity for a large system:
N2 = N1(Di1/Di2) = 600(0.192/1.12) = 103 rpm

http://www.chem.mtu.edu/~drshonna/cm4710f07/lectures/chapter10.pdf
A 10,000 liter (of liquid) bioreactor contains 5g/L of growing cells, QO2 = 20 mmol O2 / (g
cells.hr),DT = 2 m, Di = 1 m, (6 - blade turbine agitator) x 3 blades. For 1 liquid volume per minute
aeration rate (air), can the OTR(oxygen transfer rate) = OUR(oxygen uptake rate) for N = 100
rpm?
GIVEN:
VL=10,000L
QO2 = 20 mmol O2 / (g cells.hr)
DT = 2 m
Di = 1 m
(6 - blade turbine agitator) x 3 blades
N = 100 rpm
REQ’D: Is OTR = OUR?
SOL’N:
Re = Reynold's Number= ρLNDi2 / µL
ρL= 1000 kg/m3
µL =10-3 N.s/m2
Re = (1000 kg/m3)(100/60rps)(1m)2(1N/kg.m/s2) / 10-3 N.s/m2
Re = 1.67x10-6
*Np = 4(from Blanch and Clark Pmo Correlation)
Pmo = 4 (ρLN3Di5) for 1 impeller
Pmo = 4(1000 kg/m3) (100/60rps)3(1m)5
Pmo = 1.852x104kg.m2.s2/s (watts) x 3(impellers)
Pmo = 5.62x104 watts = 74.5hP
Pm: NA (aeration no.) = Qa / NDi3
NA = (10,000Lpm)(10-3m3/L) / (100min-1)(1m)3
NA = 0.10
*Pm/Pmo = 0.42(from Blanch and Clark NA Correlation)
Pm = (0.42)(5.62x104 Watts)
Pm = 2.335x104 Watts = 31.3hP
kLa (mmol O2 / ( hr.atm) = 0.60(Pm/VL(hP/103 liters))0.4 (Vs)0.5 (N,rpm)0.5
Pm/VL =31.3hP/(10)(103liters) = 3.13hP/103liters
𝜋
Vs =104 Lpm(103 cm3/L)/ 4 (2m)2(10cm/m)2 = 318.3 cm/min
kLa = 0.60(3.13)0.4 (318.3)0.5 (200)0.5

kLa = 169 (mmol O2 / (hr .atm)
OUR = X qO2 = (5g cells/ L)(20 mmol O2/g cells.hr)

OUR = 100 mmol O2/L.hr
OTR = kLa(PO2 - P*)

P * for CL = 1 mg O2/liter = HO2 CL
= (0.21 atm/(8mg O2/liter))(1 mg O2/liter)
=0.0263 atm
OTR = 169 mmoles O2/liter.hr.atm (0.21 − 0.0263) atm
OTR = 31.05 mmol O2/liter.hr
*Since OUR > OTR, we must modify the bioreactor operation in order to bring them into
balance
• increase N
• use pure O2 rather than air.
Biochemical Engineering, Second Edition by Aiba, S. et.al.
Dimensions of a fermenter equipped with two sets of standard flat-blade turbines and four baffle
plates are:
Fermenter diameter, Dt=3 m
Impeller diameter, Di=1.5 mBaffle plate width, Wb=0.3 m
Liquid depth, HL=5 m
The fermenter is used for a specific fermentation. The viscosity, µ, and the density, ρ, of the
broth are:
ρ = 1,200 kg/m3, µ = 0.02 kg/m sec
Rotation speed of impellers and aeration rate are N=60 rpm and 0.4 vvm, respectively.
Calculate:
a. Power requirements, P, for ungassed system,
b. Power requirements, Pg , when aerated,
c. Volumetric coefficient, Kv , of oxygen transfer, and
d. Hold-up, H, of bubbles.
Solution
a. Dt/Di = 3/1.5 = 2.0
HL/Di = 5/1.5 =3.33
N = 1.0 rps
𝑛𝐷𝑖2 𝜌 1 × 1.52 × 1.2 × 103
𝑁𝑅𝑒 = = = 1.35 × 105
𝜇 2 × 10−2
From Fig. 6.5,
Np = 6
𝜌𝑁 3 𝐷𝑖5 𝑁𝑃 1.2×103 ×13 ×1.55 ×6
𝑃= =
𝑔𝑐 9.81
4
= 5.57 × 10 𝑘𝑔 𝑚/ sec = 73.3 𝐻𝑝
Since the geometrical ratios, (Dt/Di)* and (HL/Di)*, of this problem deviate from Dt/Di and HL/Di
in Fig. 6.5, a correction factor, fc, which is approximately expressed as shown below will be
calculated.
𝐷 𝐻
( 𝐷𝐿 ) ∗ (𝐷𝐿 ) ∗ 2.0 × 3.33
𝑖 𝐿
𝑓𝑐= √ =√ = 0.86
𝐷 𝐻 3.0 × 3.0
( 𝐷𝐿 ) (𝐷𝐿 )
𝑖 𝐿
Then,
𝑃 ∗ = 𝑃𝑓𝑐 = 73.3 × 0.86 = 63 𝐻𝑝
If power requirements, P, with two sets of impellers can be estimated by multiplying the value of P for
one set of impellers by 2 (cf. 6.2.2.1.), the total power requirement, P** will be:
𝑃 ∗∗ = 𝑃 ∗× 2 = 63 × 2 = 126 𝐻𝑝
P**=126 Hp
b. The aeration number, Na, is calculated as follows:

𝜋 1
𝐹 0.4 × ( 4) × 32 × 5 × (60)
𝑁𝑎 = =
𝑛𝐷𝑖2 1 × 1.53
= 6.95 × 10−2
Assuming that curve A in Fig. 6.6 can be used,

Pg/P** = 0.65, Pg = P** × 0.65 = 126 × 0.65 = 82 Hp
Pg=82 Hp
c. F = 0.4 × (π/4) × 32 × 5 = 14.1 m3/min

14.1×60
𝑣𝑠 = 𝜋 = 119.7 𝑚/ℎ𝑟
( )×32
4
From Eq. (6.37).

82
𝐾𝑣 = 0.0635 × { 𝜋 }0.95 × 119.70.07
( )×32 ×5
4
= 3.45 kg mole/m3 hr atm
Supposing that the coefficient, 0.0635 be halved in the case of a flat-blade turbine (cf. Section
7.4., Chapter 7),
Kv = 1.72 kg mole/m3 hr atm
HL/Dt = 5/3 = 1.67
The correction factor, fc, is assumed as follows: (cf. Section 6.3.2.)

fc = 1.3
Then, the volumetric coefficient, Kv*, of oxygen transfer is:

Kv* = Kvfc = 1.72 × 1.3 = 2.24 kg mole/m3 hr atm
Kv* = 2.24 kg mole/m3 hr atm
It must be remembered that the above value of volumetric coefficient is maximum in terms of
oxygen transfer, because Eq. (6.37) based on the sulfite-oxidation experiment is applied in the
calculation.
d. Fig. 6.7 is used to calculate the hold-up, H.

From Fig. 6.7,
𝑃 126
( )0.4 𝑣𝑠 0.5 = { 𝜋 }0.4 × 119.70.5
𝑉 2
( 4) × 3 × 5
From an extrapolation of the solid line in Fig. 6.7,

H = 21%
Biochemical Engineering, Shigeo Katoh and Fumitake Yoshida
In an aerated stirred tank, air is bubbled into degassed water. The oxygen concentration in water
was continuously measured using an oxygen electrode, such that the data in Table 1.0 were
obtained. Evaluate the overall volumetric mass transfer coefficient of oxygen k Lɑ (in unit of per
hour). The equilibrium concentration of oxygen in equilibrium with air under atmospheric pressure
is 8.0 mg/L; the delay in response of the oxygen electrode may be neglected.
Table 1.0 Oxygen concentration in water.
Time (s) O2 concentration (mg/L)
0 0
20 2.84
40 4.63
60 5.87
80 6.62
100 7.10
120 7.40
Given:
Table 1.0
CL* = 8.0 mg/L
Required: kLɑ
Solution:
From the oxygen balance, the following equation is obtained:
dCL/dt = kLɑ (CL*- CL)
Upon integration with the initial condition CL = 0 at t = 0,
ln [CL*/ (CL*- CL)] = kLɑ (t)
Substituting values from the table; from t = 0 to t = 120
kLɑ = 0.0219/s
Calculate the power requirements, with and without aeration, of a 1.5 m-diameter stirred tank,
containing water 1.5 m deep, equipped with a six blade Rushton Turbine that is 0.5 m in diameter
d, with blades 0.25 d long amd 0.2 d wide, operating at a rotational speed of 180 r.p.m. Air is
supplied from the tank bottom at a rate of 0.6 m3/min. Operation is at room temperature. Values
of water viscosity μ = 0.001 kg/m.s and water density ρ = 1000 kg/m3; hence μ/ρ = ν = 10-6 m2/s
can be used.
Given:
Dtank = 1.5 m μ = 0.001 kg/m.s
HLiquid = 1.5 m ρ = 1000 kg/m3
N = 180 r.p.m μ/ρ = ν = 10-6 m2/s
Q = 0.6 m3/min
Required: Power requirement without aeration, Pmo; Power requirement with aeration, Pm
Solution:
The Power requirement without aeration can be obtained using Figure 1.0.
Figure 1.0 Correlation between Reynolds number (Re) and Power number (Np).
(Re) = (d2 N)/ν = (0.52 x 3)/ 10-6 = 7.5 x 105
This is in the turbulent regime. Then from Figure 1.0:
Np = 6
Pmo = 6 ρN3d5 =6(1000)33(0.5)5 = 5060 kg.m2/s3
Pmo = 5060 W
log (Pm/Pmo) = -192 (1/3)4.38(0.52 x 3/10-6)0.115(0.5 x 32/9.8)1.96/3(0.01/3 x 0.53) = -0.119
Pm/Pmo = 0.760
Hence,
Pm = 5060 x 0.760
Pm = 3850 W
A stirred-tank reactor equipped with a standard Rushton turbine of the following dimensions
contains a liquid with density ρ = 1.000 g/cm3 and viscosity μ = 0.013 g/cm.s. The tank diameter
D = 2.4 m, liquid depth HL = 2.4 m, the impeller diameter d = 0.8 m, and liquid volume = 10.85
m3. Estimate the stirred power required and the mixing time, when the rotational stirrer speed N is
90 r.p.m., that is 1.5/s.
Given:
ρ = 1.000 g/cm3 μ = 0.013 g/cm.s
D = 2.4 m HL = 2.4 m
d = 0.8 m liquid volume = 10.85 m3
N = 90 r.p.m.
Required: Pm and tm
Solution:
The Reynolds number:
Re = Nd2 ρ/ μ = (1.5 x 802 x 1)/0.013 = 7.38 x 105
Figure 1.0 Correlation between Reynolds number (Re) and Power number (Np).
From Figure 1.0, Np = 6
The power required Pm = (6 x 1.53 x 0.85 x 1000) kg m2/s3
Pm = 6650 W or 6.65 kW
Figure 1.1 Correlations for mixing times (using a standard Rushton turbine).
From figure 1.1, values of N,tm for the above Reynolds number should be about 30. Then,
tm = 30/1.5
tm = 20 seconds
A fermenter of diameter 3.6m and liquid height of 6.1 m is used for production of ustilagic acid
by Ustilagozeae. The pressure at the top of the fermenter is 1.4 atm. The vessel is stirred using
dual Rushton turbines and the fermentation temperature is 29°C. The DO tension is measured
using two electrodes: one is located near the top of the tank, the other is located near the bottom.
Both electrodes are calibrated in situ in sterile culture medium. The DO reading at the top of the
fermenter is 50% air saturation; the reading at the bottom is 65% air saturation. The fermenter is
sparged with air at 20°C at flow rate of 30 m3 min-1 measured at atmospheric pressure. Off-gas
leaving the vessel at a rate of 20.5 m3 min-1 contains 17.2% oxygen. The solubility of oxygen in
the fermentation broth is not significantly different from that in water. The density of the culture
broth is 100 kg/m3.
a. What is the oxygen transfer rate?
b. Estimate the pressure at the bottom of the tank.
c. The gas phase in large fermenters ia assumed to exhibit plug flow. Under these conditions,
no gas mixing occurs so that the gas phase composition at the bottom of the tank is equal to that
in the inlet gas stream, while the gas composition at the top of the tank is equal to that in the
outlet gas stream. For the gas phase in plug flow, estimate the oxygen solubility at the top and
bottom of the tank.
d. What is the value of kLa?
e. If the cell concentration is 16 g/L, what is the specific oxygen demand?
f. Industrial fermentation vessels are rated for operation at elevated pressures so they can
withstand steam sterilization. Accordingly, the fermenter used for ustilagic acid production can
be operated safely at a maximum pressure of 2.7 atm abs. assuming that respiration by U. zeae
and the value of kLa are relatively insensitive to pressure, what maximum cell concentration can
be supported by oxygen transfer in the fermenter after pressure is raised?
Given: Required:
DT = 3.6m a. OTR
HL= 6.1 m b. Pbottom
Qin=30 m3 min-1 c. CL*bottom, CL*top
Qout=20.5 m3 min-1 d. kLa
Ρ = 1000 kg/m3 e. Specific O2 demand
CO2,in = 50% f. CL*at Pmax = 2.7 atm abs
CO2,out = 65%
Solution:
qA
a. OTR = = Qin CO2,in - Qout CO2,out
𝑉
𝜋 𝜋
V= 4 DT2 HL = 4 (3 .6m) (6.1 m) = 62. 0904 m3
qA 1 Qin P (0.21) 𝑄𝑜𝑢𝑡 𝑃 (%𝑂2)

= 𝑅𝑉 [ − ]
𝑉 𝑇𝑖𝑛 𝑇𝑜𝑢𝑡
30 𝑚3 𝑚3
qA 1 ( )(1 atm)(0.21) (20.5 )(1.4 𝑎𝑡𝑚)(0.172)
min min
= 𝑎𝑡𝑚 [ − ]
𝑉 (0.08205 𝐿. .𝐾)(60.0904 𝑚3) (20+273) (29+273)
𝑚𝑜𝑙
𝐪𝐀 𝒎𝒐𝒍
= 𝟏. 𝟎𝟒𝟓𝟖 𝒙𝟏𝟎−𝟑
𝑽 𝑳 · 𝒎𝒊𝒏
b. PT = 1.4 atm + PL
1 𝑎𝑡𝑚
PL = ρgh = (1000 kg/m3) (9.81 m/s2) (6.1m) [101325 𝑃𝑎] = 0.5906 atm
PT = 1.4 atm + 0.5906 atm
PT = 1.9906 atm
c. Inlet at 20°C & 1 atm
𝑃02 𝑃02
[ ] = [ ]
𝐶𝐿∗ 𝑖𝑛𝑙𝑒𝑡 𝐶𝐿∗ 𝑏𝑜𝑡𝑡𝑜𝑚
1 𝑎𝑡𝑚 1 𝑎𝑡𝑚 (𝑜. 21)

[ ]= [ ]
1.38 𝑚𝑜𝑙/𝐿 𝐶𝐿∗
𝑪𝑳∗ = 𝟎. 𝟐𝟖𝟗𝟖 𝒎𝒐𝒍/𝑳
Outlet at 29°C & 1.4 atm
1 𝑎𝑡𝑚 1 𝑎𝑡𝑚 (𝑜. 21)

[ ]= [ ]
1.38 𝑚𝑜𝑙/𝐿 𝐶𝐿∗
𝑪𝑳∗ = 𝟎. 𝟐𝟖𝟒𝟏 𝒎𝒐𝒍/𝑳
𝑞𝐴
𝑉
d. kLa = 𝐶𝐿∗ − 𝐶𝐿
𝑚𝑜𝑙 1000 𝑚𝑚𝑜𝑙
1.0458 𝑥10−3 ( )
𝐿·𝑚𝑖𝑛 1 𝑚𝑜𝑙
kLa = 𝑚𝑚𝑜𝑙
(0.15)(0.2841 )− 0
𝐿
kLa = 24.5407 /min
e. Q if V=16 g/L , MW O2 = 32 g/mol
𝑚𝑜𝑙 32 𝑔
1.0458 𝑥10−3 ( )
𝐿∙𝑚𝑖𝑛 𝑚𝑜𝑙
Q= = 2.0916 x 10-3 /min
16 𝑔/𝐿
𝑔 𝑘𝑔
𝑃𝑀 (1 𝑎𝑡𝑚)(32 )( )
𝑚𝑜𝑙 1000 𝑔
ρ= = 𝐿∙𝑎𝑡𝑚 1 𝑚3
= 1.2914 kg/m3
𝑅𝑇 (0.08205 )(29+273)𝐾 ( )
𝑚𝑜𝑙∙𝐾 1000 𝐿
Sp. O2 demand= (2.0916 x 10-3/min) (1.2914 kg/m3)

Sp. O2 demand = 2.7011 x 10-3 kg/m3·min
f. Pmax = 2.7 atm abs

2.7 𝑎𝑡𝑚 (0.172) 1 𝑎𝑡𝑚
[ ]= [ 𝑚𝑚𝑜𝑙 ]
𝐶𝐿∗ 1.18
𝐿
𝑪𝑳∗ = 𝟎. 𝟓𝟒𝟖𝟎 𝒎𝒎𝒐𝒍/𝑳
Calculate the maximum possible rate of oxygen uptake at 37°C of microorganisms having a
diameter of 2/3 μm suspended in an agitated aqueous solution. It is assumed that the surrounding
liquid is saturated with 02 from air at 1 atm pressure. It will be assumed that the microorganism
can utilize the oxygen much faster that it can diffuse it. The microorganism has a density very
close to that of water.
Given:
T = 37 °C
Dp = 2/3 μm
PO2 = 1 atm
ρ= 999.9999 kg/m3
Required: Maximum O2 uptake

Solution:
Additional info: CO2 surface = 2.26 x10-4 kmol/m3 @ saturation

DAB @ PO2, inH20 = 3.25 x 10 -9 m2/s
Convective Mass Transfer over a spherical surface
ℎ𝑚 𝐷𝑝
= 2 + 0.43 (𝐺𝑇𝐴𝐵 𝑆𝐶 )0.25
𝐷𝐴𝐵
ℎ𝑚 𝐷𝑝
=2+0
𝐷𝐴𝐵
2 𝐷𝐴𝐵 2 (3.25 𝑥10−9 𝑚2 /𝑠)

ℎ𝑚 = = 2 = 9.7695 𝑥10−3 𝑚/𝑠
𝐷𝑃 x10−6 m
3
Flux O2 :
nO2 =hm (CO2; α – CO2 surface)
= (9.7695 𝑥10−3 𝑚/𝑠) (2.26 x10-4 kmol/m3 – 0)
nO2 = 2.2034 x10-6 kmol/ m2·s

A 200-L stirred fermenter contains a batch culture of Bacillus subtilis bacteria at 28°C. Air at 20°C
is pumped into the vessel at a rate of 1 vvm. The average pressure in the fermenter is 1 atm. The
volumetric flow rate of off-gas from the fermenter is measured as 189 L/min. The exit gas stream
is analyzed for oxygen and is found to contain 20.1% O2. The dissolved oxygen concentration in
the broth is measured using an oxygen electrode as 52% saturation. The solubility of oxygen in
the fermentation broth at 28°C and 1 atm air pressure is 7.8x10-3 kg/m3.
a. Calculate the Oxygen Transfer Rate
b. Determine the kLa for the system
Given: Required:
Vmed=200L a. OTR
Tmed=28°C b. kLa
Tair = 20°C
Qair = 1 vvm
P=1 atm
Qgas = 189 L/min
O2 out= 20.1% O2
Solubility of O2 @ 1 atm, 28°C = 7.8x10-3 kg/m3
Solution:
1
a. NA= 𝑉 [𝐹𝐶𝑖𝑛𝑙𝑒𝑡 − 𝐹𝐶𝑜𝑢𝑡𝑙𝑒𝑡 ]
𝐿
0.21 (1 𝑎𝑡𝑚)
CO2,inlet = 𝐿∙𝑎𝑡𝑚 = 8.7352 x10-3mol/L
(0.08205 )(20+273.15)𝐾
𝑚𝑜𝑙∙𝐾
0.201 𝑎𝑡𝑚
CO2,outlet = 𝐿∙𝑎𝑡𝑚 = 8.1386 x10-3mol/L
(0.08205 )(28+273.15)𝐾
𝑚𝑜𝑙∙𝐾
1 mol L mol
NA = 200𝐿 [(200𝐿)(1 𝑣𝑣𝑚) ( 8.7352 x10 − 3 ) − (189 min) (8.1386 x10 − 3 )]
L L
NA= 1.0442 x 10-3 mol/L.min
𝑁𝐴
b. kLa = 𝐶
𝐿∗ −𝐶𝐿
𝑚𝑜𝑙 1000𝐿 32 𝑘𝑔 𝑘𝑚𝑜𝑙 1 𝑚𝑖𝑛

(1.0442𝑥10−3 )( 3 )( )( )( )
𝐿.𝑚𝑖𝑛 𝑚 𝑘𝑚𝑜𝑙 1000 𝑚𝑜𝑙 60 𝑠
kLa = 7.8 𝑥 10−3 − (0.52)(7.8 𝑥 10−3 )
kLa = 0.1487 s-1

A bioreactor (DT = 3 m) contains 1000 L of liquid (water) and 15 g / L of growing cells whose
respiration rate is 25 mmole O2 / g cells-hr. It is being agitated by three turbine-type impellers at
25 OC and 1 atm. State where the reaction is biochemically limited or mass transfer limited for N
= 60 rpm and 1 vvm. Assume Pm/Pmo = 0.5; ρL = 1,000 kg / m3; µL = 1 x 10-3 kg/m-s. It has also
been determined experimentally that the diameter of air bubbles is 5 x 10-2 m and its velocity is
0.5 m/s.
Given:
VR = 1000 L; Cx = 15g/L; qO2 = 25 mmole O2 / g cells-hr; DT = 3 m; Pm/Pmo = 0.5; ρL = 1,000 kg /
m3; µL = 1 x 10-3 kg/m-s; Dbubbles = 5 x 10-2 m; Velocitybubbles = 0.5 m/s
Required:
Is the reaction biochemically limited or mass transfer limited?
Solution:
From James Lee Chapter 6, Di= (1/3)DT = (1/3) (3m) = 1 m
Re = Reynold's Number = ρ L N Di2 / µL
Re = (1,000 kg/m3)(60/60s)(1m)2 / (1 x 10-3 kg/m-s) = 1 x 106
From figure 9.8 of James M. Lee: Power number = 6 = Pmo / (ρL N3 Di5)
Pmo = 6 (ρL N3 Di5)
= 6 [(1,000 kg/m3)(60/60s-1)3(1m)5]
= 6000 (Watts)
Pmo (3 impellers) = 3 (6000 Watts) = 18000 Watts
Pm = 0.5 (Pmo) = (0.5)(18000 Watts) = 9000 Watts
From equation 9.71 of James Lee: kLa (s-1) = 0.026 [(Pm/VR)]0.4 (vs)0.5
Pm/VR = 9000 Watts/1m3 = 9000 Watts/m3
HT = (1m3)/ [(π/4)(3m)2] = 0.1415 m
vs = [(6)(1000 L/ min)(1min/60s)(1 m3/1000 L)( 0.1415 m)] / [(5 x 10-2 m)(0.5 m/s)(1 m3)]
= 0.566 m-1
kLa = 0.026 [9000]0.4 (0.566)0.5= 0.7466 s-1
Oxygen Transfer Rate, OTR = kLa(CL* - CL)
From table 9.1 of James Lee: CL*@pure oxygen = 1.26 mmoles O2 / L
CL* = (0.21atm)/ [(1atm)/(1.26 mmoles O2 / L)] = 0.1667 mmoles O2 / L
OTR = (0.7466 s-1)(3600s/hr) (0.1667-0) mmoles O2 / L
OTR = 448.0398 mmoles O2 / L-hr
Oxygen Uptake Rate = Cx qO2 = (15 g cells/L) (25 mmoles O2/g cells-hr)
OUR = 375 mmoles O2 / L-hr
Therefore, since OUR < OTR, the reaction is BIOCHEMICALLY LIMITED!

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PROBLEMS WITH SOLUTIONS

K1Cs ( Ceo-Ces) = k2Ces

K2Ces + k1CsCes = k1CsCeo

Rate determining equation:

−dcs dcp k3CsCeo

Substrate Concentration Initial Reaction Rate

Given: *refer to table

a. What is the maximum reaction rate?

b. What is the concentration of the substrate after 15 minutes?

Required: a.) rmax

𝑟𝑚𝑎𝑥 = 3.6254x10-6 kmol/m3 s

Cs= 2.4762 x10-4 mol/ L

a. What should be the size of a steady-state CSTR to convert 95 percent of incoming

At the second reactor ; cso =38.8650g/L

Which is more efficient

%conversion = 42.9976 % (for 2 CSTR IN SERIES)

For 1 CSTR with volume =2L

Cs= 29.1517 g/L

Conversion = 41.6969 %% ( for 1 CSTR with 2L volume)

a. Is prostigmine competitive or noncompetitive inhibitor?

Series1 Series2 Linear (Series1) Linear (Series2)

a. Prostigmine is a competitive inhibitor.

Without Inhibitor With inhibitor

r = 0.9968 μmol/L-min r = 0.9916 μmol/L-min

r = 0.9961 μmol/L-min r = 0.9876 μmol/L-min

r = 0.9781 μmol/L-min r = 0.9564 μmol/L-min

2.18 The enzyme, cathepsin, hydrolyzes L-glutamyl-L-tyrosine to carbobenzoxy-L-glutamic acid

Substrate Inhibitor Rate of CO2 Generation

Reaction [S] µM (x) 1/[S] µM-1 ν(µM.min-1) (y) 1/ν (min-1/

y-int = 1/vmax = 0.06

Kcat = Vmax / [E]T = (16 µM/min) / 0.02 µM

[I=0]: Line 1: y=4.3200×10-9x+5.0033×10-6

𝑉𝑚𝑎𝑥 = 199,868.0871 M/min 𝐾𝑚 = 8.6343×10-4 M

[I=20nM]: Line 2: y=7.4605×10-9x+5.1690×10-6

𝑉𝑚𝑎𝑥 = 193,461.0176 M/min 𝐾𝑚 𝑎𝑝𝑝 = 1.4433×10-3 M

Type of Inhibition: COMPETITIVE

REQUIRED: @T2=100 ºC ; K100 ºC = ?

Line 1: without inhibition

FOR PHENYL BUTYRATE ION:

FOR BENZOATE ION:

a. Find the rate equation for cell growth.

a. Assume Monod equation for cell growth

Applying Linear Regression

Yx/p ave= 0.2679

7 𝐿/ℎ𝑟 0.935𝐶𝑠2 𝐶𝑥2

Solve equations (1) 𝑎𝑛𝑑 (2) simultaneously

CS2 = 0.1214 g/L CX2 =5.9272 g/L

For the second condition:

𝜏𝑚 𝜇𝑚𝑎𝑥 = (1ℎ𝑟 −1 )(0.935) = 0.935

Given: rx = μmax (1 – e -CS/KS)CX

6.9 Suppose you have an organism that obeys Monod equation:

where µmax = 0.5 hr -1 and Ks = 2 g/L

𝑉 = (𝐹)( 𝜏𝑚,𝑜𝑝𝑡 ) = (100 𝐿⁄ℎ𝑟 )(2.4879 ℎ𝑟) = 248.7922 𝐿 V = 248.7922 L

Cx, opt = 20.9010 g/L

Cs2 = 0.2932 g/L

(b) For 95% substrate conversion Cs= 0.05Cso = 4.75g/l.

and the cell growth is given by;

Integrating both sides leads to;

Given: E. coli Req’d: a) µ

E. coli grows from 0.10 kg dry cell/m3 to 0.50 kg dry cell/m3 in 1 h.

Steady state Concentration in a Chemostat

F= DV= 0.26/h (60m3) = 15.6 m3h-1.