17-Apr-18

Bioreactor Design and

Configuration

Cell Growth in Continuous Culture

• Fresh nutrient is continually supplied.
• Well stirred culture.
• Products and cells are simultaneously
withdrawn.
• Steady state conditions: constant cell, product
and substrate concentration.
• Optimal environmental conditions.
• Uniform product quality.

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Cell Growth in Continuous Culture…

Turbidostat

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 17-Apr-18

Chemostat as a tool
• Evaluate KS, µmax, YX/S and other system
parameters
• Study changes in environment and effects on
cell physiology
• Select for cells with desired metabolic
capabilities (e.g. selection for cells capable of
degrading a toxic compound)

Chemostat Mass Balance ?

1. Describe dynamics of cell growth, substrate
utilization, and product formation.
2. Useful for control of bioreactors.
3. Evaluate kinetic and yield parameters.
4. Determine the optimum values for bioreactor
operating parameters.

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Ideal Constant-Stirred Tank Reactor

Chemostat

Mass Balance Statement for Cell Mass

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 17-Apr-18

Steady-State and Sterile Feed

Substrate Concentration

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 17-Apr-18

Bioreactor “Washout” Condition

• There is an upper limit on D, or the cells will
washed out of the bioreactor. The rate at
which washout occurs is termed as critical
dilution rate:

crit

With Endogenous Metabolism

• If endogenous metabolism is considered,

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 17-Apr-18

Cell Concentration
• How is X affected by D? A similar mass balance equation for S
in the absence of endogenous metabolism is written to
answer this question.

Cell Concentration (cont.)

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Effects of Endogenous Metabolism

Measurement of Maximum Cell Yield and

Maintenance using a Chemostat

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 17-Apr-18

Using a Chemostat to Determine µmax

and KS
• From data collected using a chemostat, we can
obtain the Monod Equation kinetic parameters.
• Data include S at several Dilution Rates (D), Recall
that,

Productivity of a Chemostat

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A Summary of Chemostat Response to D

Critical dilution

crit

Continuous Culture of Micro-organisms

• As we know,
 Ks 
Dop   m 1  

 K s  S0 

• Optimum biomass production

– Substitute Dop in eqn. of X for CSTF,


X op  YXM S S 0  K s K s  S 0   K s 
• Optimum productivity,

Dop X op   mYXM S 1 

Ks 

 S 0  K s K s  S 0   K s


 K s  S0 

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 17-Apr-18

Continuous Culture of Micro-

organisms
• If S0 >> Ks then, the maximum productivity can be
achieved as,
Dop X op   mYS0
• For batch fermentation, the cell productivity is given
by the ratio of the biomass produced to the batch
cycle time.
– Typically, the batch cycle time is,
tb = tg + tu Unproductive time:
• Vessel cleaning
• Sterilization
1 X 
tg  ln m  •Lag time
m  X 0  •Harvesting time

Continuous Culture of Micro-

organisms
• Thus,
Xm  X0
Productivity for batch fermenter 
tb
YXM S S 0

1 X 
ln m   t u
m  X 0 
• The ratio of the CSTF productivity to that of the batch
fermenter is,
 1 X  
 mY XM S S 0  ln m   t u 

Productivity Ratio   m  X 0    ln X m 
  tu  m
YXM S S 0 X 
 0 

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 17-Apr-18

Continuous Culture of Micro-

organisms
• The CSTF is more productive than batch fermenter if:
– Productivity > 1 (i.e., when Xm/X0 > 2.72) regardless of tu.
– Typical range of Xm/X0 is 8-10.

Problem-1

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Governing Equations

Problem-2

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Solution: problem-2
a)

b)

c)

Stirred Tank Reactor with Recycle of

Biomass
• Ex. Waste-water Treatment
– High dilution rate

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Stirred Tank Reactor with Recycle of

Biomass

Stirred Tank Reactor with Recycle of

Biomass

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Stirred Tank Reactor with Recycle of

Biomass
Assumptions: No biochemical reaction or biomass growth in the
separator (i.e. S0=S)
• The material balance over the fermenter vessel,
material  material in  formation by  output 
       
in with   with recycle  biochemical   to   Accumulation
fresh feed stream  reaction  separator 
       

• For the substrate,

dS
F0 S 0  FR S  RsV   F0  FR S  V
dt
• For the biomass,
0 (sterile feed) 0 (steady state)
dX
F0 X 0  FR X R  RX V   F0  FR  X  V
dt

Stirred Tank Reactor with Recycle of

Biomass
• For biomass balance,
FR V F
X R  X X  R X 0
F0 F0 F0
where,
FR F
 Recycle Ratio, R & 0  Dilution rate, D
F0 V
On substitution,
X
 X 1  R   X R R  0
D
The concentration ability of the separator, ξ, can be defined as,
XR

X

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Stirred Tank Reactor with Recycle of

Biomass
Division by X gives,

 1  R  R   0
D
on rearranging,

D
1  R   1
• If the separator has any concentration ability, then, D  
• If R = 0 or   1 , the equation corresponds to CSTF at steady
state with no recycle.
• Assuming Monod kinetics applies, then,
m S
D
1  R  1K s  S 

Stirred Tank Reactor with Recycle of

Biomass
• The condition for washout with recycle of biomass
can be obtained by putting SS0 as D  Dcrit,
Dcrit S0

m 1  R  1K s  S 0 

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 17-Apr-18

Problem-1

Two CSTF in Series

cannot be a
sterile feed

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 17-Apr-18

Two CSTF in Series

• For sterile feed in tank 1 X0=0,
D1 K s
S1 
 m  D1
• From the definition of yield,
 DK 
X 1  Y S 0  1 s 
  m  D1 
• Making material balance for the second vessel,
dX 2
FX 1   2V2 X 2  FX 2  V2
dt

Two CSTF in Series

• At steady state this balance gives,
2 X 2
D2 
X 2  X1
• Assuming Monod kinetics follows,
m S2 X2
D2 
K s  S2 X 2  X1
• Based on the definition of yield,

X 2  Y S 0  S 2 

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 17-Apr-18

Two CSTF in Series

• Substituting X2 in eqn. of D2 gives,

D2 
m S2 S 0  S 2 
K s  S 2  D1 K s 
  S 2 
  m  D1 
• Solving this for S2 gives,
  2
 m  D2 S 22   D1D2 K s  D2 K s   m S 0 S 22  D1D2 K s  0
  m  D1    m  D1 
Consider only the root of the eqn. with positive S2
value.

N-CSTF in Series

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 17-Apr-18

N-CSTF in Series
• In general the material balance for biomass over the
ith tank will give:
dX i
FX i 1  FX i  Vi X i  V
dt
• At steady state, the above eqn. can be rearranged to
give, DX i 1
Xi 
D  i
• For the last tank in series, where i=N, the biomass
concentration is given by,
DX N 1
XN 
D  N

N-CSTF in Series
• Similarly for the penultimate tank, where, i=N-1,
DX N  2
X N 1 
D   N 1
• Substituting XN-1 from above in eqn. of XN gives,
D 2 X N 2
XN 
D   N D   N 1 

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N-CSTF in Series
• With repeated substitution, the effluent
concentration for N growth tanks may be expressed
as, D N 1 X 1
XN  N
 D   k 
k 2

• Note that the above expression does not involve the

feed to the fermenter, which may be operated under
sterile feed condition.

N-CSTF in Series
• For sterile feed, with equal size tanks in series, the
washout condition is analogous to the single
fermenter,
Dcrit S0

m K s  S0
• Washout the first tank in the series will inevitably
result in washout in each successive vessel.

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 17-Apr-18

Stirred-tank Fermenters in Series with

Recycle of Biomass

Stirred-tank Fermenters in Series with

Recycle of Biomass
• The material balance for the biomass over the ith
fermenter gives:
dX i
F0 1  R  X i 1  F0 1  R  X i   i X iV  V
dt

• At steady state, the eqn. reduces to,

D1  R 
 X i  X i 1   
i
Xi
• Rearranging for Xi gives,
DXi 1
Xi 
D1  R  i

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 17-Apr-18

Stirred-tank Fermenters in Series with

Recycle of Biomass
• Similarly the biomass concentration for Nth tank,
D N 1 X1
XN  N
D1  R   i 
2

• In this case, even if the feed is sterile , the biomass

concentration entering the first tank is not zero and
is represented by,
FR R
X 0'  X R  XR
FR  F0 R 1

Stirred-tank Fermenters in Series with

Recycle of Biomass
• The biomass concentration for Nth tank, can be
developed further, as,
R
DN X R
XN 
R  1
N

 D1  R    
k 1
k

• For incipient washout in any tank, k  crit as D  Dcrit

• Therefore,
N R
Dcrit XR
XN 
R  1
Dcrit 1  R   crit N

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Stirred-tank Fermenters in Series with

Recycle of Biomass
• Or using, XR and rearranging to give D:

XN
 crit
Dcrit 
  R 1 N 
1  R 1    
 1  R  

• The washout criteria is typified by Sk  S0 then, if

Monod kinetics are assumed, then,
m S0
 crit 
K s  S0

Stirred-tank Fermenters in Series with

Recycle of Biomass
• The critical dilution rate can then be given as,
 m S0
K s  S0
Dcrit 
  R 1 N 
1  R 1    
  1  R  

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Problem-1
Two continuous stirred-tank fermenters are placed 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 l/h and the
substrate concentration in the fresh speed 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: Problem-1
Part-A Part-B
• S1=0.309 g/l • Dcrit=0.244 h-1< D=0.36 h-1
• X1=1.97 g/l • Final conc. same as if only
• X2=2.1+0.42S2 100 lit vessel existed,
• 0.128S22+1.379S2- S=0.309 g/l
0.01555=0 X=1.97 g/l
• S2=0.0113 g/l
• X2=2.1 g/l

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Problem-2

Plug-Flow Fermenters

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Plug-Flow Fermenters
• The material balance over the elementary volume
gives:
material  Formation  material
entering  by  leaving 
     
     Accumulation
by bulk  biochemical by bulk 
flow  reaction  flow 

• The biomass balance over elemental volume,

acdZ FA 0
F0  FI X  RX dV  F0  FI  X  dX   dX dV
dt

Plug-Flow Fermenters
• The eqn. then becomes,
R X ac dZ  FA dX  0
• The performance of the fermenter can be given by the
integral of, X Z
e
dX ac ac Residence
X RX FA 0
 dZ 
F A
Z  v time of the
liquid in the
A
fermenter
• The equation is same as for the batch fermenter but
with replacing t with v and altered boundary
conditions.

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 17-Apr-18

Plug-Flow Fermenters
• The boundary condition at the entry point is,
F X  FI X I
XA  0 0
F0  FI
• At the exit point of the reactor X=Xe. Assuming
Monod kinetics the eqn. results in,

K sY  S AY  X A  ln X e   K sY 
ln
S AY 
  v
 m S AY  X A   
 X A   m S AY  X A   S AY  X A  X e 

• The similar expression can be obtained for the

substrate concentration.

Plug-Flow Fermenters
• The material balance for the substrate results in,
Rs ac dZ  FA dS  0
and integrating,
Se
dX
R
SA s
v

where SA is given by:

F0 S 0  FI S I
SA 
F0  FI

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 17-Apr-18

Plug-Flow Fermenters
• Assuming Monod growth kinetics and
constant yield,

K sY  S AY  X A  ln1  Y S A  Se    K sY  Se 
 m S AY  X A   X    X  S Y   S   v
ln
 A  m A A  A

Plug-flow Fermenter with Recycle of

Biomass

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Plug-flow Fermenter with Recycle of

Biomass
• The biomass balance gives,
acdZ 0
F0  FR X  RX dV  F0  FR  X  dX   dX V
dt
• With substitution the equation becomes,

R X ac dZ  F0  FR dX  0
or

R X ac dZ  F0 1  R dX  0
where, FR
R
F0

Plug-flow Fermenter with Recycle of

Biomass
• The performance of the reactor will then be
expressed as,
Xe Z
dX a dZ
X RX  0 F0 1c  R 
A

• Similarly for the substrate,

Se Z
dS a dZ
S Rs  0 F0 1c  R 
A

• The substrate concentration at the point of mixing

can be given as,
S 0 F0  S R FR  S A F0  FR 

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Plug-flow Fermenter with Recycle of

Biomass
• Thus, the exit concentration of the substrate is,
S0  Se R
SA 
1  R 
• Similarly for the biomass,
X 0 F0  X R FR  X A F0  FR 
• Or if, X R  X e , then the boundary condition can be
generated as,
R
X A  Xe
1 R

Plug-flow Fermenter with Recycle of

Biomass
• The set of equations obtained are nonlinear and can
only be solved analytically for a special case. i.e.
assuming the average biomass concentration along
the length.
• The material balance for the substrate over the
reactor element may then be written as,
F0  FR S  Rs dV  F0  FR S  dS   dS V
dt
• Defining, recycle ratio R = FR/FO ; then at steady state
the above eqn. becomes.
 F0 1  R dS  Rs dV  0

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Plug-flow Fermenter with Recycle of

Biomass
• Since, dV=acdZ, then:
 F0 1  R dS  Rs ac dZ  0
• For Monod kinetics and constant yield the reaction
rate with respect to the substrate Rs may be replaced
by,  SX
Rs   m A

Y K s  S 
to give,
  m SX A ac
F0 1  R dS  dZ
Y K s  S 

Plug-flow Fermenter with Recycle of

Biomass
• Since S=SI when Z=0 and S=Se at the exit, then for a
reactor of length Z,
Se
K s  S dS    m X Aac Z dZ
 S
SA
YF0 1  R  0
which becomes,
 
S I  Se   K s ln S I    m X Aac Z
 S e  YF0 1  R 
Substituting by the definition of yield,
Xe  XI
Y
S I  Se

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Plug-flow Fermenter with Recycle of

Biomass
It may be shown that,
 
 X e  X I    m X A ac Z  YK s ln S I 
F0 1  R   Se 
For the validity of the assumption, the value of Xe and
XI should be small.

// Thanks //

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