Continuous Bioreactors –
Chemostat with Recycle
ChEE 481a
Batch Reactors
• Cell Growth
dX S
rX X m X
dt KS S
• Substrate Utilization
dS X m S X
rS
dt YX / S K S S YX / S
Continuous Reactors
• Chemostat - CSTR - continuous stirred tank reactor for
the cultivation of cells.
– mixing supplied by impellers and rising gas bubbles
– assume complete mixing - composition of any phases do not
vary with position
– liquid effluent has the same composition as the reactor
contents
Mass Balance on Chemostat
Acc = in - out + gen - cons
dci
VR Fcif Fci VR r fi
dt
– VR - reactor volume
– F - volumetric flow rate of feed and effluent streams (they are
equal)
– ci - concentration of component i in the reactor
– cif - concentration of component i in the influent or feed stream
• If we have a steady state reactor - no changes in
composition with time
then dcand
i
0
dt
F
rfi (ci cif )
VR
• Define as the dilution rate - F 1
D
VR
– reciprocal of the mean holding or residence time
– detention time
• For cell mass, if we assume a sterile feed:
ci = X and Xf = 0 and
D=m at SS
rx = mX then mX = DX
Chemostat with Monod Kinetics
mS KS
D S
KS S mm 1
• The above equations only holds if tmmmax >1
• If tmmmax < 1 or D>m
– washout of the cells occurs
– Cells leave the reactor faster than they are dividing.
mS f
Dmax Near washout the reactor is very sensitive to
KS S f variations in D
• Small change in D large shifts in X and/or S
• If mmax = 0.5 hr-1 then D< 0.4 hr-1
Substrate Balance on Chemostat -
Intracellular Product
dci
VR Fcif Fci VR r fi
dt
• If ci = S
1 dS
FS 0 FS VR X VR
YX / S dt
• At Steady State
X
D S 0 S X YX / S S 0 S
YX / S
• With Monod
Ks D
X YX / S S 0
m D
Class Exercise
• Problem 6.17
• E. coli is cultivated in continuous culture under
aerobic conditions with glucose limitation. When
the system is operated at D= 0.2 hr-1, determine the
effluent glucose and biomass concentrations
assuming Monod kinetics (S0 = 5 g/l, mm= 0.25 hr-1 ,
KS = 100 mg/L, Y x/s = 0.4 g/g)
Chemostat with Recycle
FR, XR
F, X0 F, X2
V, X1
F+FR, X1
• F - nutrient flow rate
• V - reactor volume
• X1 - x concentration in reactor
• X2 - X concentration in effluent
• XR - X concentration in recycle
•
Chemostat with Recycle Cell
mass equation
Acc = in - out + gen
dX 1
F X0 + FR XR - (F+ FR) X1 + VmX1 = V
dt
FR, XR
F, X0 F, X2
V, X1
F+FR, X1
Chemostat with Recycle cont.
Define Substitutions
a = FR/F • F + FR = (1 + a)F
recycle ratio • FRXR term
C = XR /X1 FR = Fa
concentration factor
XR = CX1
FRXR = aCFX1
dX 1
F X0 + FR XR - (F+ FR) X1 + VmX1 = V
dt
dX 1
F X0 + aCFX1- (1 + a)F X1 + VmX1 = V
dt
Recycle cont
• Assume
dX 1 Chemostat can be
– steady state =0
dt operated at higher
dilution rates than the
– sterile feed X0 = 0
specific growth rate
Then when cell recycle is
(aC - 1 -a)F + Vm = 0 used.
If D = F/V for recycle
m = D(1+ a(1 -C))
if C > 1 (concentration of cells) then a(1 - C) < 0
then m < D
Substrate balance - Recycle
X 1 dS
FS 0 FS V (1 ) FS V
YX / S dt
• At Steady state and substituting for m
D YX / S ( S 0 S )
X 1 YX / S ( S 0 S )
(1 C )
Recycle Substrate cont.
• Assuming Monod
K S D(1 C )
S
max D(1 C )
YX / S K S D(1 C )
X1 S0
(1 C ) max D(1 C )
Fed Batch Reactor
• Reactor Design Equation
V dN A
FA0 FA rA dV
dt
• No outflow FA = 0
• Good Mixing rA dV
term out of the integral
dN A d C A V
FA0 rA V
dt dt
Fed Batch Continued
• Convert the mass (NA) to concentration. Applying
integration by parts yields
dC A dV
FA0 rAV V CA
• Since dt dt
dV
FA0
• Then dt
dC A
FA0 rAV V C A FA0
• Rearranging dt
dC A FA0 C A FA0
rA
dt V V
Fed Batch Continued
dC A FA0
• Or 1 C A rA
dt V
• Used when there is substrate inhibition and
for bioreactors with cells.
Fed-batch Reactors
d (Vci )
Vr fi F (t )c fi
dt
dV
F (t )
dt
Differentiation the above equation using chain rule, and substitute
for dV/dt
dci F (t )
rfi [cif ci ]
dt V
Fed-batch cont.
• Cell balance – sterile feed
rfi X
KS D
dX S
( D) X m D
dt
• This can be a steady state reactor if substrate is
consumed as fast as it enters (quasi-steady-state).
Then dX/dt = 0 and m = D, like in a chemostat.
Recall, D = F / V
Fed batch cont
• Substrate balance – no outflow (Fcout = 0), sterile feed
• St = SV and Xt = XV (mass of substrate or cells in reactor at a
given time)
• S0 = substrate in feed stream
substrate substrate
in consumed
dS t
X t
no substrate out
Substrate FS 0 (Flow out = 0)
balance dt YX / S
t
Cell dX
balance X t
dt
Fed batch cont.
• Quasi steady state for St – change in substrate
is very small in reactor and is consumed as
rapidly as fed
X t
FS 0
YX / S
t
dX
X FYX / S S 0
t
Integrating from t=0 to t
dt
X X 0 FYX / S S 0t
t t
Fed batch cont.
• Quasi steady state for St – change in substrate
is very small in reactor and is consumed as
rapidly as fed (then, dSt/dt = 0)
X t
FS0
YX / S
YX / S FS0
X
t
t
dX
X t
FY X S
/S 0 Integrating from t=0 to t
dt
XX
t
0
t
FYX S
/S 0t Note, "t" was missing.
Fed batch cont.
X X FYX / SS0 t
t t
0
• What this means
– the total amount of cells in the reactor increases
with time
– dilution rate and m decrease with time in fed
batch culture
– Since m = D, the growth rate is controlled by
the dilution rate.
Product profiles in fed batch
• Product profiles can be obtained by using the definitions
of Yp/s or qp.
• When Yp/s is constant (at quasi-steady-state with S <<S0):
P ≈ Yp/s S0, and
FP ≈ FYp/s S0,
• When qp is constant,
t
dP
q pX t
dt
• where Pt is the total amount of product in culture
• Substituting
X X FYX / SS0 t
t t
0
• yields t
dP
q p X m V0 Ft
dt
Ft
P P0 q p X m V0 t
t t
2
• in terms of product concentration
V0 V0 Dt
P P0 q pX m t
V V 2
Class Exercise – 9.4
• Penicillin is produced in a fed-batch culture with the
intermittent addition of glucose solution to the culture
medium. The initial culture volume at quasi-steady state
is V0= 500 L, and the glucose containing nutrient
solution is added with a flow rate of F = 50 L/h. X0 = 20
g/L, S0 = 300 g/L, mm = 0.2 h-1, Ks = 0.5 g/L and Y x/s=
0.3 g/g
• Determine culture volume at t = 10 h
• Determine concentration of glucose at t = 10 h
• Determine the concentration and total amount of cells at
t = 10 h
• If qp = 0.05 g product.g cells h and P0 = 0.1 g/L,
