Chemostat

10.37 Chemical and Biological Reaction Engineering, Spring 2007 Prof. K.
Dane Wittrup Lecture 13: Biological Reactors- Chemostats This lecture covers: theory of the chemostat, fed batch or semi-continuous fermentor operations
Biological Reactors (Chemostat)

Concentration/Combustion constant Biological CSTR [S]0 F
[S], x
V
F Figure 1. Diagram of a chemostat. F = Volumetric flow rate
x=
biomass volume
[S]0 = Concentration of growth limiting substrate. (for growing cells) At steady-state, biomass balance In Out + Prod = Acc Sterile feed: In=0 Steady state: Acc=0
Fx + rx V = 0 at steady-state Cell growth kinetics rx = x Fx + x V = 0 F Solve = V F 1 = D=Dilution rate V
=D
Biological Mechanical
Cite as: K. Dane Wittrup, course materials for 10.37 Chemical and Biological Reaction Engineering, Spring 2007. MIT OpenCourseWare (http://ocw.mit.edu), Massachusetts Institute of Technology. Downloaded on [DD Month YYYY].
When at steady-state, can control cell mass. Allows precisely reproducible cell states. Not easy to run at steady-state. Material balance on [S] (sugar concentration) In Out + Prod = Acc 0 at steady-state
F [ S ]0 F [ S ]
1 x V = 0 Yx
s
Yield coefficient
mass biomass created mass substrate consumed
Divide by V
x D ([ S ]0 [ S ]) = Yx
change in sugar concentration s
At steady-state
=D
x = Yx ([ S ]0 [ S ])
s
What is the value of
[ S ] ? What more information do we need?
= f ([ S ])
must choose a growth model to connect
and
[S ]
Monod growth model:
max [ S ]
K s + [S ]
at steady-state
D=
max [ S ]
K s + [S ]
[S ] =
Ks D max D
substitute in x equation
10.37 Chemical and Biological Reaction Engineering, Spring 2007 Prof. K. Dane Wittrup
Lecture 13 Page 2 of 6
Ks D x = Yx [ S ]0 max D s
Specifying
max , K s , Yx , D , [ S ]0 , can predict x , [ S ] .

s
x < 0 is non-physical but formally in solution
max D
can go to 0. If you turn knobs incorrectly: if D is too high, the cells cannot
grow fast enough to reach steady-state. Washout will occur. so use x = 0 to find Dmax
Dmax =
max [ S ]0
K s + [ S ]0
For D > Dmax washout, no steady-state.
x, S
mass volume
S
x
washout
Dmax
Figure 2. Biomass/volume versus dilution rate. Beyond the maximum dilution rate, washout occurs. For real systems K s << [ S ]0 . Most cell growth systems reach maximum at fairly low
concentrations; hence x is flat, then drops off sharply. If biomass is the product, is there a best operating condition? What should we consider?
dx optimize x with respect to D? D=0 (no, because this would be batch reactor) dD
Define productivity as biomass
(reactor
volume )( time )
= xD
d ( xD) = 0 for optimum. (is a maximum) dD

x
xD
D
Doptimum
Figure 3. Left: Biomass/volume versus dilution rate. Right: Productivity versus dilution rate.
Ks Doptimum = max 1 K s + [ S ]0
K s << [ S ]0
Doptimum max
Dmax
Close to washout conditions. Operability would be difficult. We would not want to run too close to washout conditions. Fed-batch fermentor (microbes or mammalian cells) -used to achieve very high cell densities (e.g. hundreds of grams cell dry weight (c.d.w)/liter) If you want x final =
100 g L wt 200 g 20% Toxic, sugar content cells will die L volume
If Yx 0.5 , [ S ]0 =
s
Why do we not feed all at once? Cells will die.
Calculate medium feed rate in order to hold
constant.
[S]0
x(t) S(t), very small

Figure 4. Diagram of a fed-batch fermentor.
If
is constant, biomass = biomass t =0 e
There is a dilution term, because as we feed in fresh medium, volume will change. Volume often doubles.
x V = x0 V0 e t
Feed
F [ S ]0 =
sugar feed
x0 V0 e t
sugar consumed
Yx s
Assume all converted into biomass.
F=
x0 V0 e t [ S ]0 Yx
s
Exponential flow rate. Typically Dilution:
specified as small.
dV =F dt
x0 V(t ) = V0 1 + e t 1) ( [ S ]0 Yx s
x=
biomass
V
t
= 1+
x0 e t x0 e t 1) ( Yx [ S ]0
s
x0 V0 e V
Logistic equation
Figure 5. Graph of logistic growth.
t
If product is something cells are making: Product synthesis kinetics 1)
1 dP = x dt
growth associated (e.g. ethanol)
P
2)
product volume not growth associated (e.g. antibiotics, proteins, antibodies) integrate for amount of product.
1 dP = x dt
P = xdt
0 t

Chemostat

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10.37 Chemical and Biological Reaction Engineering, Spring 2007 Prof. K.

Biological Reactors (Chemostat)

Fx + rx V = 0 at steady-state Cell growth kinetics rx = x Fx + x V = 0 F Solve = V F 1 = D=Dilution rate V

mass biomass created mass substrate consumed

What is the value of

[ S ] ? What more information do we need?

must choose a growth model to connect

Monod growth model:

max , K s , Yx , D , [ S ]0 , can predict x , [ S ] .

x < 0 is non-physical but formally in solution

For D > Dmax washout, no steady-state.

d ( xD) = 0 for optimum. (is a maximum) dD

Why do we not feed all at once? Cells will die.

Calculate medium feed rate in order to hold

x(t) S(t), very small

is constant, biomass = biomass t =0 e

Assume all converted into biomass.

Exponential flow rate. Typically Dilution:

Figure 5. Graph of logistic growth.

growth associated (e.g. ethanol)

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