Kinetics of Microbial Growth Factors Determining Growth

& Synthesis of Products

Unlimited growth

Assuming td = 0.33 h, • Absolute Factors

in 48 h,
one cell would become
-12
If a cell weighs 10 g,
then the total would be
Nutrients; pH; Temperature; Oxygen
43
2.33 X 10 cells
• Rate-Determining Factors
2.23 X 1031 g Temperature; pH; Mass Transfer; Energy Transfer

This would be 4000 times the weight of the earth!

Dr. Clem Kuek ZIP/Lect+Prac/IndusMicrobiol/Lectures/GrowthKinetics.doc 1 Dr. Clem Kuek ZIP/Lect+Prac/IndusMicrobiol/Lectures/GrowthKinetics.doc 2

Kinetics of Batch Culture Kinetics of Batch Culture 2

Phases of growth in batch culture

Growth Rate, rx

Deceleration

population
Maximum
Δx
=

Exponential

Death
Δt

Ln Population Size
or as t becomes infinitesimally small

Acceleration
dx
= Units e.g. g cells mL-1h-1
dt

Lag
Specific Growth Rate, μ μ Specific Growth Rate μmax
rx
= Units e.g. g g-1 cells mL-1h-1
x

Time
Dr. Clem Kuek ZIP/Lect+Prac/IndusMicrobiol/Lectures/GrowthKinetics.doc 3 Dr. Clem Kuek ZIP/Lect+Prac/IndusMicrobiol/Lectures/GrowthKinetics.doc 4
Kinetics of Batch Culture 3 Kinetics of Batch Culture 4

Integrating equation 2.1

Cell Number
The exponential phase may Exponential
gives
be described by
xt = x0 eμt
Phase Log
10000 Cell Number Exponential
(2.2)
dx 10000 Phase

= μx (2.1) 8000
where
xo = [original biomass]
dt xt = [biomass after time t]
where
e = base of natural log
x = [microbial biomass] 6000 1000

t = time (h) Taking natural log, equation 2.2

μ = specific growth rate (h-1) 4000
becomes

2000 ln xt = ln x0 + μt 100

0 Thus, plot of ln X vs. t gives

1 4 7 10 13 16
Time straight line in the exponential 10
1 4 7 10 13 16
phase, slope of which = μ Time

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Determining x from data r Determining rx from data 2

Growth Rate, rx
Time X S
(h) (g L-1) (g L-1)
0 0.100 40.00
1 0.134 39.93
2 0.180 39.83
3 0.241 39.70 1. Hand-drawn tangent
4 0.323 39.50 y
5 0.433 39.30 rx =
6 0.581 38.97 x
7 0.778 38.50
8 1.040 38.00
9 1.400 37.20
2. Numerical differentiation
Difference between values
10 1.870 36.20
on either side of data point
11 2.500 34.80
12 3.350 32.90
rx = x − x 3 1

t −t
2
13 4.490 30.50 3 1
14 6.000 27.20
15 8.000 22.80
16 10.70 17.10 3. Curve fitting
[
r x = ln x − ln x ]⋅ x
17 14.10 9.60
3 1
18 17.90 1.11
t −t
2 2
19 17.90 1.11 3 1

20 17.90 1.11

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 Determining μmax from data
Determining μ from data
1. By tabulation of values for μ through the exponential phase of the
Using the values for growth determined as described previously,
culture.
Specific Growth Rate may be estimated by the relationship
Time
(h)
x rx μ = rx/x
(g L-1) (h-1)*

rx 0 0.100 - -

μ= μmax 1
2
3
0.134
0.180
0.241
0.040
0.054
0.072
0.298
0.300
0.299
 μmax

x 4 0.323 0.096 0.297

Specific Growth Rate

5 0.433 0.129 0.298
6 0.581 0.172 0.296
7 0.778 0.230 0.296
8 1.040 0.311 0.299
However, in batch culture, 9 1.400 0.415 0.296

μ does not remain constant. 10

11
1.870
2.500
0.550
0.740
0.294
0.296
12 3.350 0.995 0.297
Substrate concentration is 13
14
4.490
6.000
1.325
1.755
0.295
0.293
a major affecting factor. 15 8.000 2.350 0.294
Substrate concentration 16 10.70 3.050 0.285
17 14.10 3.600 0.255
*rx via numerical differentiation.
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Determining μmax from data 3

Determining μmax from data 2
Compare 1
= Ks ⋅ 1 + 1 with y = mx + c
2. Lineweaver-Burke plot μ μ S μ m m

r =
dx Thus, when
1
is plotted against 1 , we get
x μ S
dt
Note large number of
μ ⋅S ⋅x 1
=
m

points in this area

K +S s μ
r μ ⋅S
=
x m

x K +S s

rx Intercept
Taking the reciprocal and since =ì 1
x =
x 1 K +S μmax Ks
= =
s

r μ μ S Slope =
x m ⋅
μmax
K1 S
= +
s

μ ⋅S μ Sm m ⋅

K 1 1 1
= ⋅ +
s

μ S μ m m
S
Dr. Clem Kuek ZIP/Lect+Prac/IndusMicrobiol/Lectures/GrowthKinetics.doc 11 Dr. Clem Kuek ZIP/Lect+Prac/IndusMicrobiol/Lectures/GrowthKinetics.doc 12
Yield factor Y Determination of Yield Factor on carbon substrate, Yx/s 2

rs rx
Thus, when x is plotted against x , we get
Y = the ratio of product or cell quantity resulting
from a certain quantity of input
r Note large number of
e.g. Yx/s Yield of cell weight per unit weight substrate utilized x
Yp/n Yield of product weight per unit weight of nitrogen utilized points in this area

Determination of Yield Factor on carbon substrate, Yx/s

rs = rs + m s
Intercept = ms 1
Slope =
Y x
x
s Yx/s
rs = 1 ⋅ rx + m s
x Y x x
s r
x
where rs = rate of consumption of carbon substrate
ms = maintenance coefficient on carbon substrate
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The Importance of μmax The Importance of the Yield Factor Y

Y indicates the degree of efficiency of the

For processes where maximal growth rates are
conversion of substrates into desired products.
diserable, attainment of μmax in culture is important.
Attainment of efficient Y translates directly into economic
Since μmax is determined by the
efficiency, and thus productivity.
• genetics of the microorganism
• conditions of culture Since Y is determined by the
Attainment of μmax has implications for both determinants. • genetics of the microorganism
• conditions of culture
For other processes, identification of μmax is • nature of the input (substrate)
important so that it can be avoided
e.g. in the production of secondary metabolites. Attainment of an efficient Y has implications for the
determinants.
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