Prof. José A.

Pérez
Cell Culture Reactor Design
Fed-Batch Operation
Applications Figure 1. Fermenter in fed-batch operation
Control of substrate concentration
Overcome of catabolic repression F, Si, Xi, Pi
Substrateinhibition due to high concentrations
High growth by oxygen demand
Examples :bakers ’ yeast and penicillin production. Fermenter

V
Total mass balance
d (ρ V )
ρ i F i − ρ o F o=
dt V0
ρi ≈ρ and F o =0
dV
F= Mass balance of substrate
dt
Mass balance of cells d (V S )
F i S i −F o S o −r S V =
d (V X ) dt
F i X i −F o X o +r X V =
dt q dS dV
F i X i +( μ− α ) X V =
d (V X)
dt
[
F i Si −
μ
Y X /S Y P/S ]
+m S + P X V =V
dt
+S
dt
qP dS
F i X i +(μ − α) X V =V
dX
dt
+X
dV
dt
D( Si −S)−
[ μ
Y X/S
+m S +
Y P/ S]X=
dt
dX
D X i +(μ− α ) X= +D X Mass balance of product
dt
d (V P)
F i P i − F o P o +r P V =
X i =0 and μ ≫ α dt
dX dP
μ X= +D X D( P i −P)+q P X=
dt dt
1
 Fed-Batch Operation

dV dX qP dS dP
F=
dt
; D( X i − X )+(μ− α ) X=
dt
; D (S i −S)−
μ
[
Y X/S
+m S +
Y P/ S]X=
dt
; D ( Pi − P)+q P X =
dt

One can imagine the situation where the fermenter initially is operated batch until reaching a high cell concentration
and the substrate is virtually exhausted rapidily. Under this conditions, intermitent fed is started with fresh medium
and volumetic flow F. Then, biomass concentration almost unswitched and all fed substrate is drained inmmediately,
therefore:

dX dS dP
≈ 0 ; S≪S i ⇒ ≈0; ≈0 ; that is known as pseudo−steady state or quasi− steady state .
dt dt dt

D( X i − X)+(μ− α ) X≈0; If μ ≫ α and X i =0

μ≈ D
μ S
Monod model : D≈μ = m
KS+ S
KS D
S≈
μ m− D
Operative constraint : μm− D≠0⇒ μm > D

Only cells production or product directly linked with energy generationand maintenance requirements negleted :
μ dS
D( Si −S)− X = ≈0 ⇒ X≈Y X / S S i
Y X/S dt
dP
D ( Pi − P)+q P X = ≈0 ⇒ P≈Y P / S S i
dt
2
 Fed-Batch Operation

Cellular mass balance

d X̄ dV dX
X̄= X V ; =X +V
dt dt dt
dX d X̄
≈0 ; = X F ; X ≈Y X /S Si
dt dt
d X̄= F Y X / S S i d t
X̄= X̄ 0 + F Y X / S S i t fb ; t fb : time for fed−batch operation.

Product mass balance

d P̄
=q P X̄ ; P̄=P V
dt
d P̄
=q P X̄ 0 +q P F Y X / S S i t
dt
2
t fb
P̄− P̄ 0 =q P X̄ 0 t fb +q P F Y X /S Si
2
X̄ P̄
X 0= 0 ; P 0 = 0
V0 V0
2
V0 V0 t fb
P=
V( ) ( )
P0 + q X t +q D X
V p 0 fb p 2
V
( ) γ= 0
V
2
t fb
P= γ P 0 + γ q p X 0 t fb +q p D X
2
3
Problem 2.
Penicillin is produced by Penicillium chrysogenum in a fed-batch culture with
the intermittent addition of glucose solution to the culture medium. The initial
culture volume is V0 = 200 L, and glucose-containing nutrient solution is added
with a flow rate of F = 30 L/h. Glucose concentration in the feed solution and
the initial concentrations in the reactor are S0 = 300 g/L, Si = 1 g/L, and X0 = 20
g/L. The kinetic and yield factors of the organism are µm = 0.2 h-1, KS = 0.5 g/L,
and YX/S = 0.3 g-dw/g-glucose. Using the pseudo-steady state approximation,
we determine:

1. The culture volume at t = 24 h.

2. The concentration of glucose at t = 24 h.
3. The concentration and total amount of cells when t = 24 h.
4. The product (penicillin) concentration in the vessel at 24 h, if m qP = 0.05 g-
product/(g-cells h) P0 = 0.1 g/L.