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Bioreactor Modelling-A-Lecture-6
Bioreactor Modelling-A-Lecture-6
Bioreactor
dX
___ vX
dt
The proportionality constant P is called
the specific growth rate. (units are time-1)
Notes:
b) Modified Monod
PMAX S
P = ______________ S0 = Initial Substrate
KS + KS0S0 + S Concentration
PMAX S
_________ 1
c) Contois P= Predicts P __ at low S
KSXX + S X
Predicts P ĺ 0 at high X
PMAX S _______
_______ KI
d) Monod-like “Noncompetitive P=
Inhibition” KS + S KI + I
PMAX S _______
_______ KP
e) Monod-like “Noncompetitive P=
Product Inhibition” KS + S KP + P
or
dS
_ ___ = mSX
dt
Maintenance
2. Cell mass
The substrate may be used for the production
of new cellular components which ultimately
become new cells. The rate of substrate
consumed to produce more cells is
proportional to the rate of new cells produced:
dS
_ ___ dX
v ___
dt dt
Cells
_ Y dS
___ dX
X/S
= ___
dt dt
Cells
or
_ dS
___ 1 dX
= ____ ___
dt YX/S dt
Cells
3. Products
The substrate may be used for the synthesis of
chemical products. These are usually at the
termination of biochemical pathways.
dP
___
Volumetric rate of product formation = QP =
dt
1 ___
___ dP
Specific rate of product formation = qP =
X dt
There are several relationships which can
exist between the rate of product formation
(qP) and the specific rate of substrate
consumption (qS) or specific growth rate (P).
specific product formation rate (qP)
_ dS
___ dP
YP/S = ___
dt dt
Products
_ dS
___ 1 ___
____ dP
=
dt YP/S dt
Products
dP
___ dX
v ___
dt dt
dP
___ dX
___
= YP/X
dt dt
Note: dP 1 ___
___ dX
___ = YP/X X
dt X dt
QP = YP/X P X growth
associated
qP = YP/X P products
b. Non-growth associated
The specific rate of product formation is a
constant.
qP = E
QP = EX
c. Mixed-growth associated
qP = DPE Luedeking-Piret Equation
QP = DPX EX
d. My personal preference, but a lot of work
qP = f(P)
qP = f(qS)
4. Summary
Thus, the total substrate utilization may be
written:
dS
_ ___ dS
_ ___ dS
_ ___ dS
_ ___
=
dt dt dt dt
Total Maint Cells Products
dS
_ ___ PX
____ QP
- rS = = m SX + + ____ = QS
dt YX/S YP/S
Total
QS
qS = ___
X
C. General material balances
1. Derivation
FOUT
b) Limiting substrate:
d(VS)
_____
= FINSIN - FOUTSOUT + rSV
dt
c) Product:
d(VP)
_____
= FINPIN - FOUTPOUT + rPV
dt
d) Water:
d(VW)
_____
= FINWIN - FOUTWOUT + rWV
dt
2. Common simplifications
WIN = WOUT = W
and insignificant water is generated
rW = 0
b) Reactor is well-mixed
POUT = P
SOUT = S
XOUT = X
PIN = 0; XIN = 0
rX = PX
Results of these simplications:
d(VX)
_____
a) Cells: = - FOUTX + PXV
dt
d(VS)
_____
b) Substrate: = FINSIN - FOUTS + rSV
dt
d(VP)
_____
c) Product: = - FOUTP + rPV
dt
dV
____
d) Water: = FIN - FOUT
dt
D. Batch operation
1. Introduction
A batch process is a closed culture which occurs
when FOUT = 0 and FIN = 0.
dX
____
a) Cells: = + PX
dt
dS
____
b) Substrate: = + rS
dt
dP
____
c) Product: = + rP
dt
dX
____ PMAXSX _______
________ N
So, =
dt KS + S KN + N
dS
____
Energy substrate: = + rS
dt
In order to use this equation, we need an expression for
rS, which we have derived previously.
PX
____ QP
- r S = m SX + + ____
YX/S YP/S
We will consider the case where no product is
formed (QP = 0). Incorporating two-substrate Monod
model into substrate equation:
dS
____ PMAXSNX
= – mSX – __________________
dt YX/S(KS + S)(KN + N)
Non-energy substrate: dN
____
= + rN
dt
A “substrate” model may be used for every nutrient.
However, typically no other nutrient (like N) is directly
incorporated into products (QP = 0), and no other
nutrient except oxygen is used for maintenance (mN =
0). Thus,
PX
____
- rN =
YX/N
dN
____ PMAXSNX
= – __________________
dt YX/N(KS + S)(KN + N)
In summary
dX
____ PMAXSNX
= _______________
dt (KS + S)(KN + N)
dS
____ PMAXSNX
= – mSX – __________________
dt YX/S(KS + S)(KN + N)
dN
____ PMAXSNX
= – __________________ Note
dt YX/N(KS + S)(KN + N) dX dN
___ v ___
dt dt
3. Simulation
Parameters:
mS = 0.05 g/gh *
KS = 10 mg/L = 0.01 g/L* (S = glucose)
KN = 0.01 g/L ‡
PMAX = 0.80 h-1
YX/S = 0.44 g/g *
YX/N = 8.0 g/g †
Simulated Cases:
Case I: “Carbon-Limited”
X0 = 0.005 g/L S0 = 30 g/L N0 = 5 g/L
† unpublished data
‡ yeast; Cramer et al. 2002
* see subsequent pages
Case I: “Carbon-Limited”
S 0.6
120 20 10
P (h-1)
100 Oxygen Limited
8
15 0.4
80
6
60 10
4
40 N 0.2
5 2
20
0 0 0 0.0
0 2 4 6 8 10 12 14
Time (h)
30 0.8
140 14
S
Substrate and N (g/L)
120 25 12
Cell Density (g/L)
OUR (mmol/Lh)
80 8
15 0.4
60 6
10
40 4 0.2
20 5 2
N
0 0 0 0.0
0 2 4 6 8 10 12 14
Time (h)
4. Conclusions:
E. Chemostat operation
F
___
Define: Dilution rate = D = [time-1]
V
The material balances become….
dX
____
a) Cells: = - DX + PX
dt
dS
____
b) Substrate: = D(SIN - S) + rS
dt
dP
____
c) Product: = - DP + rP
dt
a) Cells: DX = PX
b) Substrate: D(SIN - S) = - rS
c) Product: DP = rP
or D=P
!
This equation states that the cells’ specific
growth rate is determined by the dilution rate.
qPX
Commonly the term ____ is taken to be small:
YP/S
PX
- rS = mSX + ____
YX/S
The material balance becomes:
DX
____
D(SIN – S) = mSX +
YX/S
D(SIN – S)
__________ D
or = mS + ____
X YX/S
X
________ OBS
Define: = YX/S
(SIN – S)
D
_____ D
____
So: OBS = mS +
YX/S YX/S
3. Biomass yield
D
_____ D
____
OBS = mS +
YX/S YX/S
OBS
This equation really describes YX/S = f(D).
mS and YX/S are parameters. Note that the
observed biomass yield is a function of dilution
rate, but the true biomass yield is not. The
equation may be written:
YX/S
OBS
YX/S
OBS
At high dilution rate YX/S Æ YX/S
4. Calculation of maintenance coefficient and true
biomass yield
y = mx + b
1 1
_____
Plot ___ (“x”) versus OBS (“y”)
D YX/S
1
Intercept = _____
YX/S
Slope = mS
2) Hofstee Plot
Rewrite equation as
OBS
OBS – mSYX/SYX/S
______________
YX/S = + YX/S
D
y = mx + b
OBS
YX/S
Plot _____
OBS
(“x”) versus YX/S (“y”)
D
Intercept = YX/S
This plot is
almost never
Slope = – mSYX/S used
y = mx + b
D
____
Plot D (“x”) versus OBS (“y”)
YX/S
Intercept = mS
1
_____
Slope =
YX/S
Comments:
YX/S is sometimes referred as maximum yield
coefficient or true yield coefficient.
5. More on maintenance
D
_____ D
____
Becomes: OBS = 0.05 +
YX/S 0.44
D YX/S
0.05 0.306 Assuming
0.10 0.361 maintenance is
constant…
0.20 0.396
0.40 0.417
0.80 0.428
What is maintenance?
Protein turnover in E. coli during stationary phase is
5% per hour (Mandelstam & McQuillen, 1968). On
the basis of a cell composition of 50% protein, 4 ATP
needed per amino acid residue, an average
molecular mass of an amino acid of 100, and 24
ATP generated per mole glucose, the glucose
consumption rate necessary to sustain protein
turnover is:
0.05 0.5
_____ 4 mol ATP
g protein _____________
__________ mol AA
_________
h g cells mol Amino Acid 100 g AA
180 g glucose
mol glucose ___________
___________
= 0.0076 g glucose/gh
24 mol ATP mol glucose
What is maintenance?
6. Product DP = rP
So, DP = qPX
7. Calculating substrate concentration in chemostat
PMAXS
________
P=
KS + S
Thus, at steady-state (“SS”) we would expect:
P S
________
D = MAX SS
KS + SSS
Or: DKS
________
SSS =
PMAX - D
DKS
________ (0.40 h-1) (10 mg/L)
__________________
SSS = = = 10 mg/L
PMAX - D (0.80 h-1 – 0.40 h-1)
D(SIN – S)
__________ D
= mS + ____
X YX/S
DKS
________
SSS =
PMAX - D
We can therefore directly estimate the steady-
state biomass concentration:
[ DKS
D SIN – ________
PMAX - D
XSS = _____________________
]
D
mS + ____
YX/S
SIN
XSS § ______________
mS
____ 1
____
+
D YX/S
SIN
XSS § ______________
mS
____ 1
____
+
D YX/S
DKS
________
SSS =
PMAX - D
1
____ KS ____
____ 1 1
= + ____
D PMAX SSS PMAX
y = mx + b
1 1
Plot ___ (“x”) versus ___ (“y”)
SSS D
1
Intercept = _____
PMAX
KS
_____ The slope is actually
Slope = close to zero
PMAX because KS is so
small.
10. Industrial use of a chemostat
Disadvantages:
Specifically:
1. Wastewater Treatment
2. Biomining
11. Why conduct a laboratory chemostat experiment?
a) Dilution Rate
F
D = ___
V
0.155 L/h
D = ________ = 0.155 h-1
1.00 L
b) Residence Time
1 1
_______
T = ___ = = 6.45 h
D 0.155 h-1
X 2.45 g/L
YX/G = ________ = ___________________
OBS
X 2.45 g/L
YX/A = ________ = _______________
OBS
= 3.90 g glucose/Lh
QG 3.90 g/Lh
qG = _____ = ___________
X 2.45 g/L
= 1.59 g glucose/g cells h
Note: An interesting calculation can be made from the
value of qG:
2 u 10-13 g cells h
u ________________ u ______
cell 3600 s
= 2.77 g pyruvate/Lh
QP 2.77 g/Lh
qP = _____ = ___________
X 2.45 g/L
= 1.13 g pyruvate/g cells h
i) Oxygen
Inlet:
xN + xO + xCO2 = 1
xN + 0.2092 + 0 = 1
xN = 0.7908
IN PQGAS
______ (1000)(1 atm)(1.00 L/min)
__________________________
nTOTAL = =
RT (0.08206 L atm/molK)(273.15K)
i) Oxygen (cont’d)
IN
nTOTAL = 44.61 mmol/min
IN
nN = (0.7908)(44.61) = 35.28 mmol/min
IN
nO = (0.2092)(44.61) = 9.33 mmol/min
Outlet:
xN + xO + xCO2 = 1
xN + 0.2013 + 0.0067 = 1
IN OUT
xN = 0.7920 nN = nN = 35.28 mmol/min
i) Oxygen (cont’d)
OUT OUT
nTOTAL = nN / xN = 35.28/0.7920 = 44.55 mmol/min
OUT
nO = (0.2013)(44.55) = 8.97 mmol/min
OUT
nCO2 = (0.0067)(44.55) = 0.30 mmol/min
IN OUT
OUR = (nO - nO )/V = [(9.33 – 8.97) mmol/min]/1.00 L
OUT IN
CER = (nCO2 - nCO2)/V = [(0.30 – 0.00) mmol/min]/1.00 L
CER
______
RQ =
OUR
17.91 mmol/Lh
______________
RQ = = 0.816 mol/mol
21.96 mmol/Lh
l) Carbon Balance
Carbon Generated
CH1.595N0.263O0.374P0.023K0.013S0.006
FW = 24.70 g DCW/mol
Notes:
Carbon Consumed
Carbon Balance
= 95.1%
dX
____
a) Cells: = - DX + PX
dt
dS
____
b) Substrate: = D(SIN - S) + rS
dt
Simplifications/Assumptions:
PX qPX
- rS = mSX + ____ + ____
YX/S YP/S
qPX
Again take the term ____ to be small:
YP/S
PX
- rS = mSX + ____
YX/S
dX
____ PMAXSX
________
= – DX +
dt KS + S
dS
____ PMAXSX
= D(SIN – S) – mSX – ___________
dt YX/S(KS + S)
Experimental Conditions:
SIN = 10 g/L
D = 0.30 h-1
Simulated Initial Conditions: (Note XSS = 4.1 g/L and SSS = 6 mg/L)
0.40
0.35
Specific Growth Rate (h )
-1
0.30
0.25
0.20
Case I
0.15 Case II
Case III
Case IV
0.10
0 1 2 3 4 5 6
2 Case I
Case II
Case III
Case IV
0
0 1 2 3 4 5 6
Case I
Case II
Case III
Biomass Concentration (g/L)
6 Case IV
0 1 2 3 4 5 6
Residence Time
to reach within 2%
of steady-state
X S P
Case I 3.33 3.87 3.42
Case II 3.06 3.54 3.09
Case III 2.22 2.73 2.28
Case IV 3.90 4.38 3.93
Conclusions:
dXƍ
____ PMAXƍSXƍ
________ Note:
= – DXƍ + A change in either KSƍ or PMAXƍ (to KSƎ or
dt KSƍ + S PMAXƎ) will affect the values of Xƍ and XƎ.
The parameters mSƍ or YX/Sƍ appear in the
dXƎ
____ PMAXƎSXƎ
________
differential equation for the substrate
concentration (S). These parameters do
= – DXƎ +
dt KSƎ + S not separately, directly affect values of Xƍ
and XƎ.
dS
____ PMAXƍSXƍ
___________
= D(SIN – S) – mSƍXƍ –
dt YX/Sƍ(KSƍ + S)
PMAXƎSXƎ
– mSƎXƎ – ___________
YX/SƎ(KSƎ + S)
There is only one substrate concentration (S), but two strains
contribute to its consumption.
Parameters:
mS = 0.05 g/gh
KS = 10 mg/L = 0.01 g/L
PMAX = 0.80 h-1
YX/S = 0.44 g/g
Experimental Conditions:
SIN = 5 g/L
Initial Conditions:
D0 = 0.30 h-1
X0 = 2.047 g/L (steady-state at D = 0.30 h-1)
S0 = 6.00 mg/L (steady-state at D = 0.30 h-1)
Simulated Cases:
Case I:
at t = 5 h (1.5 RT), a new strain appears with
10% lower KS.
original
strain competitor
(ƍ) (Ǝ)
X 2.046 0.001 g/L
mS 0.05 0.05 g/gh
KS 0.010 0.009 g/L
PMAX 0.80 0.80 h-1
YX/S 0.44 0.44 g/g
PMAXS
________ (0.80)(6.0)
________ (0.80)(6.0)
________
P= Pƍ = = 0.30 h-1 PƎ = = 0.32 h-1
KS + S 10.0 + 6.0 9.0 + 6.0
Case I: 10% lower KS.
2.5 0.33
6.0
0.32
2.0
P (h-1)
5.6
0.30
1.0
5.4 0.29
5.2 0.5
0.28
Case II:
at t = 5 h (1.5 RT), a new strain appears with
6.7% greater PMAX.
original
strain competitor
(ƍ) (Ǝ)
X 2.046 0.001 g/L
mS 0.05 0.05 g/gh
KS 0.010 0.010 g/L
PMAX 0.80 0.853 h-1
YX/S 0.44 0.44 g/g
PMAXS
________ (0.80)(6.0)
________ (0.853)(6.0)
__________
P= Pƍ = = 0.30 h-1 PƎ = = 0.32 h-1
KS + S 10.0 + 6.0 10.0 + 6.0
Case II: 6.7% greater PMAX.
2.5 0.33
6.0
0.32
2.0
P (h-1)
5.6
0.30
1.0
5.4 0.29
5.2 0.5
0.28
Case III:
at t = 5 h (1.5 RT), a new strain appears with
10% lower mS.
original
strain competitor
(ƍ) (Ǝ)
X 2.046 0.001 g/L
mS 0.05 0.045 g/gh
KS 0.010 0.010 g/L
PMAX 0.80 0.80 h-1
YX/S 0.44 0.44 g/g
P (h-1)
5.6 0.315
1.0 0.310
5.4
0.305
5.2 0.5
0.300
Case IV:
at t = 5 h (1.5 RT), a new strain appears with
10% greater YX/S.
original
strain competitor
(ƍ) (Ǝ)
X 2.046 0.001 g/L
mS 0.05 0.05 g/gh
KS 0.010 0.010 g/L
PMAX 0.80 0.80 h-1
YX/S 0.44 0.484 g/g
P (h-1)
5.6 0.315
1.0 0.310
5.4
0.305
5.2 0.5
0.300
dS
____ PMAXƍSXƍ
___________
= D(SIN – S) – mSƍXƍ –
dt YX/Sƍ(KSƍ + S)
PMAXƎSXƎ
– mSƎXƎ – ___________
YX/SƎ(KSƎ + S)
Conclusions:
FOUT
FIN FIN
Rigid
FOUT
“pipe”
Load Cell
By mass By volume
FIN FOUT
FpH FAF
FIN
FOUT
Nutrient Feed (FIN) + Feed of base/acid (FpH) + Feed of anti-foam (FAF) + etc.
FIN
FOUT
a) Lowersubstrate concentration?
• less cost of (all) nutrients
• lower O2 demand
• less cells
• less O2 consumed and CO2 generated, so
measurement of O2 and CO2 must be more sensitive.
Note that, according to Monod model, changing inlet
concentration SIN changes XSS but not SSS
FIN FOUT
F. Accelerostat
1. Motivations
2. Simulation
dS
____ PMAXSX
= D(SIN – S) – mSX – ___________
dt YX/S(KS + S)
dD
____
=K
dt
We will start this simulation at a steady-state of
D = 0.05 h-1 which is our initial value for the third
differential equation.
Parameters: Experimental Conditions:
mS = 0.05 g/gh SIN = 5 g/L
KS = 10 mg/L = 0.01 g/L
PMAX = 0.80 h-1
YX/S = 0.44 g/g
Initial Conditions:
D0 = 0.05 h-1
X0 = 1.528 g/L (steady-state at D = 0.05 h-1)
S0 = 0.67 mg/L (steady-state at D = 0.05 h-1)
Simulated Cases:
0.05 h-1 Æ 0.40 h-1
Case I: K = 0.002 h-2 175 h
Case II: K = 0.01 h-2 35 h
Case III: K = 0.02 h-2 18 h
Conclusions:
1. Introduction
The growth limiting substrate is the substrate which is
consumed according to the intrinsic biological kinetics of
the cells, and therefore it controls the cell growth. Simply
put, the stoichiometry of cell growth dictates the quantity of
energy and elements needed, and the growth limiting
substrate is the nutrient exhausted (first) in the formation of
cells. Note that other, non-limiting substrates are
consumed at their maximal specific rate.
N-Limited Chemostat
C-Limited Chemostat
Batch
qacetate
q S*
Specific glucose consumption rate (qS)
Steady-state (chemostat)
Steady-state (chemostat)
YX/S
NADH/NAD
{ qA { qA
Steady-state (chemostat)
glucose-6P
fructose-6P
PEP
pyruvate
acetyl CoA
Steady-state (chemostat)
¡ YX/S NADH/NAD
z qA z qA
glucose-6P
fructose-6P
PEP
pyruvate
acetyl CoA
2 1.5 2
gltA 0.8
1
acnA 1.0
1
0.4 0.5
0 0.0 0
0.0
-0.5
-1 -0.4 -1
-1.0 acnB icd
-2 -0.8 -1.5 -2
1.5 1.5 2 2
1.0 sucA 1.0 sucB sucC 1
sucD
1
Expression Ratio
0.5
0.5
0.0 0 0
0.0
-0.5
-0.5 -1 -1
-1.0
-1.0 -1.5 -2 -2
wild-type
3 3 3 3
2
1
sdhA 2 sdhB 2
1
sdhC 2
1
sdhD With NADH oxidase
1
0 0 0
0 -1
-1 -1
-2 -1 -2 -2
-3 -2 -3 -3
3 0.5 2 1.5
fumA fumB 1 fumC 1.0 mdh
2
0.0 0.5
0
1
-1 0.0
-0.5
0 -2 -0.5
-1 -1.0 -3 -1.0
0.3 0.6 0.9 1.2 1.5 0.3 0.6 0.9 1.2 1.5 0.3 0.6 0.9 1.2 1.5 0.3 0.6 0.9 1.2 1.5
3. Nitrogen limitation
Nitrogen-limited growth in a chemostat alters metabolism
compared to carbon-limited growth. The obvious
difference between these two experimental conditions is
that the steady-state (carbon) substrate concentration is
much greater in an N-limited culture.
N (residual N)
C-limited N-limited
C (residual)
Regime Regime
Ratio C:N
No
Increasing C Æ
Transition zone
The transition zone shifts to higher C:N ratio with lower dilution rate
because of higher relative maintenance (necessitating more C).
Egli and Zinn, 2003
a) Oxygen Transfer
OUR = OTR
ȝX
____
mOX + = kLa(c*O2 – clO2)
YX/O
ȝX
_____
We examined our
0.014X + = 180(8.0 – 0.0)(0.001) lab’s 2.0 L fermenters,
(1.87) and the kLa obtained
is about 180 h-1
b) Heat Transfer
c) Products
d(VX)
_____
= – FOUTX + PXV
dt
d(VS)
_____
= FINSIN – FOUTS + rSV
dt
d(VP)
_____
= – FOUTP + rPV
dt
dV
____
= FIN – FOUT
dt
d(VX)
_____
= PXV
dt
d(VS)
_____
= F(t) SIN + rSV
dt
d(VP)
_____
= rPV
dt
dV
____ Obviously, volume
= F(t) is not constant
dt
How should process be operated to achieve a
constant P = PC?
a) Find F(t)
d(VX)
_____
= PCXV From cell balance
dt
XV t
d(VX)
_____
= PC dt
(VX)
X0V0 0
XV = X0V0exp(PCt) EQN. A
Note that X = f1(t)
and V = f2(t)
d(VS)
_____
= F(t) SIN + rSV From substrate balance
dt
VdS SdV
or - rSV = F(t) SIN - ____ - ____
dt dt
( )
PC
or for P = PC - rS = X mS + ____
YX/S
( )
PC
F(t) SIN = XV mS + ____
YX/S
( )
PC
F(t) SIN = X0V0exp(PCt) mS + ____
YX/S
X0V0
F(t) = _____
SIN ( PC
mS + ____
YX/S ) exp(PCt)
dV
____
= F(t) From ‘water’ balance
dt
dV
____
or = Dexp(PCt)
dt
1
___
V - V0 = [ Dexp(PCt) - Dexp(PCt0) ]
PC
}
1
D
___
V - V0 = [ exp(PCt) - 1 ]
PC
X0V0
_____
SIN (
mS +
PC
____
YX/S
V = V0 + __________________
)
[ exp(PCt) - 1 ]
PC
X0
where E ____
SIN ( mS
____
PC
1
+ ____
YX/S ) Dimensionless
c) Find X(t)
Recall XV = X0V0exp(PCt)
X0V0exp(PCt)
X(t) = ___________
V(t)
X0exp(PCt)
X(t) = ________________
1 + E[exp(PCt) - 1@
X0
where E ____
SIN ( mS
____
PC
1
+ ____
YX/S )
3. Example calculations
Data:
V0 = 1.5 L
X0 = 2.0 g/L
SIN = 500 g/L
mS = 0.05 g/gh
YX/S = 0.44 g/g
E = 0.0104
3. Example calculations (cont’d)
2 exp(0.15t)
X(t) = _______________________ [g/L]
0.9896 + 0.0104 exp(0.15t)
60
2.4
50
Cell Density (g/L)
2.2
X
Volume (L)
40
2.0
30
1.8
V 20
1.6 10
1.4 0
0 5 10 15 20 25
Time (h)
2. Volume
V(t) = V0 + FCt
3. Biomass
d(VX)
_____
= PXV
dt
dV dX
X ___ + V ___ = PXV
dt dt
dX dV
V ___ = PXV – X ___
dt dt
dX dV
V ___ = PXV – X ___
dt dt
dX
___
dt
( dV
1 ___
= X P – __
V dt
)
dX
___
dt
( FC
= X P – _______
V0 + FCt )
Using Monod model to relate specific growth rate
and substrate concentration:
dX
___
dt
(PMAXS
= X ________
KS + S
FC
– _______
V0 + FCt ) Solve
numerically
4. Substrate
d(VS)
_____
= FCSIN + rSV
dt
P
dV dS
S ___ + V ___ = FCSIN – X
dt dt (
mS + ____
YX/S )
dS
___
dt
1
[
= __ FCSIN – X
V (
P
mS + ____
YX/S )]
Solve
numerically
Comments:
5. Example calculations
V0 = 1.5 L
X0 = 2.0 g/L
SIN = 500 g/L
mS = 0.05 g/gh
YX/S = 0.44 g/g
PC = 0.15 h-1
2 exp(0.15t)
X(t) = _______________________ [g/L]
0.9896 + 0.0104 exp(0.15t)
X0 = 20 g/L
t0 = 16.01 h (“time = 0” for the constant phase)
FC = 12.97 mL/h = 0.01297 L/h
V0 = 1.657 L
PCKS
________ (0.15h-1)(0.01 g/L)
_________________
S0 = = = 0.0023 g/L
PMAX – PC (0.80 h-1 – 0.15 h-1)
Coupled equations to solve numerically
V(t) = V0 + FCt
dS
___
dt
1
[
= __ FC(SIN – S) – X
V (
P
mS + ____
YX/S )]
dX
___
dt
PMAXS
= X ________
KS + S
( FC
– _______
V0 + FCt )
PMAXS
________
P=
KS + S
1.9 50 0.14
80
Cell Density (g/L)
OUR (mmol/Lh)
0.12
Volume (L)
1.8 40
60
P (h-1)
0.10
1.7 30
40 0.08
1.6 20
0.06
20
1.5 10 0.04
0 1.4 0 0.02
0 5 10 15 20 25 30
Time (h)
Practical Matters:
Process OURMAX X
C-limited Batch 170 mmol/gh 12.8 g/L
N-limited Batch 91 mmol/gh 8.0 g/L
Exp/Const Fed-Batch 59 mmol/Lh 46 g/L
Comments:
E. H. Battley (2003) Absorbed heat and heat of formation of dried microbial biomass. J. Therm
Anal Calorim 74:709-721.
M. Dauner, T. Storni, U. Sauer (2001) Bacillus subtilis metabolism and energetics in carbon-
limited and excess-carbon chemostat culture. J. Bacteriol. 183:7308-7317.
T. Egli, M. Zinn (2003) The concept of multiple-nutrient-limited growth of microorganisms and its
application in biotechnological processes. Biotechnol. Adv. 22:5-43.
I. S. Farmer, C. W. Jones (1976) The energetics of Escherichia coli during aerobic growth in
continuous culture. Eur. J. Biochem. 67:115-122.
W. Harder, L. Dijkhiuzen (1976) In: Continuous Culture 6: Applications and New Fields, A. C.
R. Dean, D. C. Ellwood, C. G. T. Evans, J. Melling eds. Ellis Horwood Ltd., Chichester.
References
A. Kayser, J. Weber, V. Hecht, U. Rinas (2005) Metabolic flux analysis of Escherichia coli in
glucose-limited continuous culture. I. Growth-rate-dependent metabolic efficiency at steady
state. Microbiology 151:693-706.
J. Mandelstam, K. McQuillen. 1968. Biochemistry of bacterial growth. 2nd ed. Blackwell Sci.
Publications Ltd. Oxford.
S. J. Pirt (1965) The maintenance energy of bacteria in growing cultures. Proc. Royal Soc.
London Ser. B. 163:224-231.
P. van Hoek, J. P. Van Dijken, J. T. Pronk (1998) Effect of specific growth rate on fermentative
capacity of Baker’s yeast. Appl. Environ. Microbiol. 64:4226-4233.