Cell Growth Models

Lecture 6
Fall 2007
Quantifying Growth Kinetics
Structured vs Unstructured -
S - model divides the cell mass into components
U - assumes fixed cell composition - exponential
growth phase of batch or in continuous
Segregated vs Nonsegregated
S - assumes different types of cells exist
N - all cells are the same type

Unstructured, Nonsegregated Models
(Monod)

Assumptions
One limiting substrate
Semi empirical relationship
Single enzyme system with
Michaelis-Menten kinetics is
responsible for the uptake of
substrate
Amount of enzyme is
sufficiently low to be growth
limiting
Low population density
Most commonly used
expression for growth

S K
S
S
m
+
=

m
- maximum growth rate when
S >> K
s
K
s
- saturation constant -
concentration of the rate-limiting
substrate when the specific rate
of growth is equal to one half of
the maximum.
K
s
= S when = 1/2
m

In general =
m
for S >>
K
s
and for S << K
s

S
m
K
S
=
Cell Growth - Monod
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0 5 10 15 20 25
Time (hr)
C
e
l
l

C
o
n
c
e
n
t
r
a
t
i
o
n

(
g
/
L
)
Does not
account for
death phase
Does not
account for
lag phase
Unstructured, Nonsegregated Models
Other models - Section 6.3.2.1 in text
assuming one limiting substrate
Blackman
Tessier
Moser
Contois
Multiple Substrates are growth limiting
Usually do not use unstructured, nonsegregated
model
Inhibition Models
(Very similar to enzyme models)
Substrate Inhibition
High substrate concentration inhibits growth
If a single-substrate enzyme catalyzed reaction is the
rate-limiting step then inhibition of enzyme activity
results in inhibition of microbial growth.
Noncompetitive Substrate Competitive Substrate


|
|
.
|

\
|
+
|
.
|

\
|
+
=
I
S
m
K
S
S
K
1 1

S
K
S
K
S
I
S
m
+
|
|
.
|

\
|
+
=
1

Inhibition Models - cont.

Product Inhibition
High concentrations of product can be inhibitory
Underlying mechanism of product inhibition is unknown
Approximated as exponential or linear decay functions
Noncompetitive Product Competitive Product

|
|
.
|

\
|
+
|
.
|

\
|
+
=
P
S
m
K
P
S
K
1 1

S
K
P
K
S
P
S
m
+
|
|
.
|

\
|
+
=
1

Inhibition Models - cont.

Inhibition by Toxic Compounds
inhibition of growth is analogous to enzyme inhibition
Noncompetitive Competitive



Uncompetitive Cell Death
|
|
.
|

\
|
+
|
.
|

\
|
+
=
I
S
m
K
I
S
K
1 1

S
K
I
K
S
I
S
m
+
|
|
.
|

\
|
+
=
1

|
|
.
|

\
|
+
|
|
.
|

\
|
+
+
=
I I
S
m
K
I
S
K I
K
S
1
) / 1 (

'
d
S
m
k
S K
S

+
=

Batch Reactors
Cell Growth


Substrate Utilization


Product (cometabolic contaminants use negative sign)

X
S K
S
X
dt
dX
r
S
m
X
+
= = =

S X S
m
S X
S
Y
X
S K
S
Y
X
dt
dS
r
/ /
+
= = =

|

o
| o
+
+
= =
+ = = =
S K
S
dt
dP
X
q
Y
dt
dP
X
q
S
m
p
g g X P p
1
1
/
Logistic Equation
Batch Growth
Equation
Combines Batch
growth, Monod and
Yield Coefficients
No maintenance
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0 5 10 15 20
Time (hr)
C
e
l
l

C
o
n
c
e
n
t
r
a
t
i
o
n

(
g
/
L
)
Rate Expression for Growth
(1)

Yield Expression
(2)
Substituting into Eq. 1 for S from Eq 2:


Integrating

Logistic Equation
X
S K
S
X
dt
dX
r
S
m
X
+
= = =

) (
0 / 0
S S Y X X
S X
=
X
X X S Y Y K
X X S Y
dt
dX
S X S X S
S X m
) (
) (
0 0 / /
0 0 /
+ +
+
=

| | t S Y X X S Y
X S Y
Y K
X
X
X S Y
X S Y Y K
m S X S X
S X
S X S
S X
S X S X S
= +
+

|
|
.
|

\
|
+
+ +
0 / 0 0 /
0 0 /
/
0 0 0 /
0 0 / /
/ ) ( ln
) (
ln
) (
) (
Describes
Sigmoidal
shape batch
growth curve
Unstructured, Nonsegregated Models
Disadvantage of Unstructured, Nonsegregated Models
No attempt to utilize or recognize knowledge about cellular
metabolism and regulation
Show no lag phase
give no insight to the variables that influence growth
assume a black box
assume dynamic response of a cell is dominated by an
internal process with a time delay on the order of the
response time
most processes are assumed to be too fast (psuedo ss) or too
slow to influence the observed response.

Filamentous Organisms
Types of Organisms
mold
bacteria or yeast entrapped in a spherical gel particle
formation of microbial pettlets in suspension
Model - no mass transfer limitations

R - radius of the cell floc or pellet or mold colony
Then the growth of the biomass (M)can be written as



const k
dt
dR
= =
3 / 2
2 2
4 4
M
dt
dM
or
R k
dt
dR
R
dt
dM
p

t t
=
= =
3 / 1
) 36 ( t
p
k =
Where:
Filamentous Organisms - cont.
Integrating the equation:



M
0
is usually very small then
Model is supported by experimental data.


3 3
3 / 1
0
3 3
|
.
|

\
|
~
|
.
|

\
|
+ =
t t
M M

3
t M
Chemically Structured Models

Improvement over nonstructured, nonsegregated models
Need less fudge factors, inhibitors, substrate inhibition, high
concentration different rates etc.
Model the kinetic interactions amoung cellular
subcomponents
Try to use Intrinsic variables - concentration per unit cell
mass- Not extrinsic variables - concentration per reactor
volume
More predictive
Incorporate our knowledge of cell biology

In Class Exercise
The results shown below correspond to typical
batch culture dynamics. Calculate the following:
A)
max
assuming Monod growth
B)

Y
X/S

C) o and | assuming mixed growth and
non-growth associated product formation