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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
MichaelisMenten 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
+
=
µ
µ
µ
m
 maximum growth rate when
S >> K
s
• K
s
 saturation constant 
concentration of the ratelimiting
substrate when the specific rate
of growth is equal to one half of
the maximum.
• K
s
= S when µ = 1/2µ
m
• In general µ = µ
m
for S >>
K
s
and for S << K
s
S
m
K
S µ
µ =
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 singlesubstrate enzyme catalyzed reaction is the
ratelimiting 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
µ
µ
S
K
S
K
S
I
S
m
+


.

\

+
=
1
µ
µ
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
µ
µ
S
K
P
K
S
P
S
m
+


.

\

+
=
1
µ
µ
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
µ
µ
S
K
I
K
S
I
S
m
+


.

\

+
=
1
µ
µ


.

\

+


.

\

+
+
=
I I
S
m
K
I
S
K I
K
S
1
) / 1 (
µ
µ
'
d
S
m
k
S K
S
÷
+
=
µ
µ
Batch Reactors
• Cell Growth
• Substrate Utilization
• Product (cometabolic contaminants use negative sign)
X
S K
S
X
dt
dX
r
S
m
X
+
= = =
µ
µ
S X S
m
S X
S
Y
X
S K
S
Y
X
dt
dS
r
/ /
+
÷ = ÷ = =
µ µ

µ
o
 oµ µ
+
+
= =
+ = = =
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
+
= = =
µ
µ
) (
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 /
÷ + +
÷ +
=
µ
  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
µ = ÷ +
+
÷


.

\

+
+ +
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
¸
µ t t µ
=
= =
3 / 1
) 36 ( tµ ¸
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) µ
max
assuming Monod growth
• B)
Y
X/S
• C) o and  assuming mixed growth and
• nongrowth associated product formation
Quantifying Growth Kinetics • Structured vs Unstructured – S .exponential growth phase of batch or in continuous • Segregated vs Nonsegregated – S .model divides the cell mass into components – U .assumes fixed cell composition .all cells are the same type .assumes different types of cells exist – N .
Unstructured.saturation constant concentration of the ratelimiting substrate when the specific rate of growth is equal to one half of the maximum. • Ks = S when = 1/2m • In general = m for S >> Ks and m S for S << Ks KS . Nonsegregated Models (Monod) mS • Assumptions – One limiting substrate – Semi empirical relationship – Single enzyme system with MichaelisMenten kinetics is responsible for the uptake of substrate – Amount of enzyme is sufficiently low to be growth limiting – Low population density KS S • Most commonly used expression for growth m .maximum growth rate when S >> Ks • Ks .
2 0.3 0.Cell Growth .6 0.5 0.1 0 0 5 10 15 20 Does not account for death phase Does not account for lag phase 25 Time (hr) .7 Cell Concentration (g/L) 0.4 0.Monod 0.
2.Unstructured. Nonsegregated Models • Other models .1 in text assuming one limiting substrate – – – – Blackman Tessier Moser Contois • Multiple Substrates are growth limiting – Usually do not use unstructured.Section 6. nonsegregated model .3.
Inhibition Models (Very similar to enzyme models) • Substrate Inhibition – High substrate concentration inhibits growth – If a singlesubstrate enzyme catalyzed reaction is the ratelimiting step then inhibition of enzyme activity results in inhibition of microbial growth. Noncompetitive Substrate m KS 1 S S 1 K I Competitive Substrate m S S K S 1 K I S .
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 m KS 1 S P 1 K P Competitive Product m S P K S 1 K S P .
cont.Inhibition Models . • Inhibition by Toxic Compounds – inhibition of growth is analogous to enzyme inhibition Noncompetitive m KS 1 S I 1 K I Competitive m S I K S 1 K I S Uncompetitive m S Cell Death KS I (1 I / K ) S 1 K I I mS KS S ' kd .
Batch Reactors • Cell Growth rX S dX X m X dt KS S • Substrate Utilization m S X dS X rS dt YX / S K S S YX / S • Product (cometabolic contaminants use negative sign) 1 qp X 1 qp X dP YP / X g g dt m S dP dt KS S .
2 0.3 0.4 0.Logistic Equation 0.1 0 0 5 10 15 20 • Batch Growth Equation • Combines Batch growth.7 0. Monod and Yield Coefficients • No maintenance Time (hr) Cell Concentration (g/L) .6 0.5 0.
Logistic Equation Rate Expression for Growth mS dX rX X X dt KS S (1) Yield Expression X X 0 YX / S ( S 0 S ) Substituting into Eq. 1 for S from Eq 2: (2) m (YX / S S 0 X 0 X ) dX X dt ( K S YX / S YX / S S 0 X 0 X ) Integrating Describes Sigmoidal shape batch growth curve ( K S YX / S YX / S S0 X 0 ) X K S YX / S ln (Y S X ) ln (YX / S S0 X 0 X ) / YX / S S0 mt (YX / S S0 X 0 ) X /S 0 0 X0 .
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. Nonsegregated Models • Disadvantage of Unstructured.
no mass transfer limitations dR k const dt – R .radius of the cell floc or pellet or mold colony dM dR 4R 2 k p 4R 2 dt dt or dM M 2 / 3 dt Then the growth of the biomass (M)can be written as Where: k p (36 ) 1/ 3 .Filamentous Organisms • Types of Organisms – mold – bacteria or yeast entrapped in a spherical gel particle – formation of microbial pettlets in suspension • Model .
3 . • Integrating the equation: 1/ 3 t t M M0 3 3 3 3 • M0 is usually very small then M t • Model is supported by experimental data.Filamentous Organisms .cont.
Not extrinsic variables . high concentration different rates etc. nonsegregated models • Need less fudge factors.Chemically Structured Models • Improvement over nonstructured. inhibitors.concentration per reactor volume • More predictive • Incorporate our knowledge of cell biology . substrate inhibition. • Model the kinetic interactions amoung cellular subcomponents • Try to use Intrinsic variables .concentration per unit cell mass.
Calculate the following: • A) max assuming Monod growth • B) YX/S • C) and assuming mixed growth and • nongrowth associated product formation .In Class Exercise • The results shown below correspond to typical batch culture dynamics.
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