Growth Kinetics Calculation

Cell Growth Kinetics
- Introduction
- Growth patterns and kinetics in batch
culture
- growth phases
- effect of factors: oxygen supply
- heat generation
- Growth kinetics (Monod Equation)
- Growth in continuous culture (ideal
chemostat)
Growth Kinetics
Introduction
- Autocatalytic reaction: The rate of growth is directly related to
cell concentration
substrates + cells extracellular products + more cells
S + X P + nX
S: substrate concentration (g/L); X: cell mass concentration (g/L);
P: product concentration (g/L); n: increased number of biomass.
Net specific growth rate (1/time):
t: the time
1 dX
net
X dt
Growth Kinetics
Introduction
Net specific growth rate (1/time):
net g k d
g :
Gross specific growth rate (1/time)
kd :
The rate of loss of cell mass due to cell death

or endogenous metabolism
Endogenous metabolism: during the stationary phase,

the cell catabolizes cellular reserves for new building blocks and
for energy-producing monomers.
Growth Kinetics
Introduction
Net specific replication rate (1/time):
1 dN
N dt
R
R
N
'
kd :
'
kd
: Cell number concentration (cell number /L)

: Gross specific replication rate (1/time)
The rate of cell death (1/time)
Growth Kinetics
Introduction
- Quantifying cell concentration:
- direct: no suspended solid and interference
compounds.
cell mass concentration preferred
dry weight, optical density (turbidity) (600-700nm
Wave Length)
cell number density:
Petroff-Hausser slide (hemocytometer), plate counts,
etc.
Growth Kinetics
- Quantifying cell concentration:
- indirect: direct method is inapplicable. (mold solid
state fermentation)
Cell mass can be determined by measurement
of protein, DNA or ATP. e.g. 1mg ATP/g dry weight
bacterial cell.
If 100 mg ATP/L is measured, then the cell mass:
100 mg (ATP/L)/1 mg ATP/g dry cells=100 (g dry
weight cells/L)
Growth Kinetics
- Growth patterns and kinetics in batch culture
- growth phases
In batch culture:
- lag phase
- logrithmic or exponential growth phase
- deceleration phase
- stationary phase
- death phase
Typical growth curve for a bacterial population
Batch Growth Kinetics

Lag phase
A period of adaptation for the cells to their
new environment
New enzymes are synthesized.
A slight increase in cell mass and volume, but no
increase in cell number
Prolonged by low inoculum volume, poor
inoculum condition (high % of dead cells), age of
inoculum, nutrient-poor medium
Multiple lag phases: (diauxic growth) medium
contains more than one carbon source
Diauxic growth

Exponential growth phase
In this phase, the cells have adjusted to
their new environment and multiply
rapidly (exponentially)
Balanced growth all components of a cell
grow at the same rate.
Growth rate is independent of nutrient
concentration, as nutrients are in excess.

The balance of cell mass in a batch culture
gives:
dX
net X , X X 0 at t 0
dt
Integration of the above equation yields :
X
ln
net t , or X X 0e net t
X0
X and X 0 are cell concentrations at time t and t 0
The slope net is constant.

net R m
m is the maximum specific growth rate (1/time)
Doubling time of cell mass: the time required to double
the microbial mass:
ln X / X 0
ln 2 0.693
d
net
net
net

Deceleration growth phase
Very short phase, during which growth decelerates
due to either:
Depletion of one or more essential nutrients
The accumulation of toxic by-products of growth
(e.g. Ethanol in yeast fermentations)
Period of unbalanced growth: Cells undergo
internal restructuring to increase their chances of
survival

Stationary Phase:
With the exhaustion of nutrients (S0) and build-up
of waste and secondary metabolic products
- The growth rate equals the death rate.
- There is no net growth in the organism population.
- Cells may have active metabolism to produce secondary
metabolites.
Primary metabolites are growth-related: ethanol by S.
cerevisae.
Secondary metabolites are non-growth-related:
antibiotics, pigments.
Kinetic Pattern of Growth and

Product Formation
Growth-associated Mixed-growth-associated Non growth-associated

Stationary phase
- Cell lysis may occur and viable cell mass may drop.
A second growth phase may occur and cells may grow on
lysis products of lysed cells (cryptic growth)
- Endogenous metabolism occurs by catabolizing cellular
reserves for new building blocks and energy-producing
monomer (maintenance energy).
The rate describing the conversion of cell mass into
maintenance energy or the loss of cell mass due to cell
lysis:
dX
kd X
dt
k d is the rate constant for endogenous metabolism.

Death Phase:
The living organism population decreases with time,
due to a lack of nutrients and toxic metabolic byproducts.
The rate of death usually follows:
dN
'
k d N
dt
'
k d is the first - order death rate constant.

Yield coefficients: defined based on the amount of
consumption of another material.
X
Growth yield : Y
X /S
S
P
Product yield : Y
P/S
S
Growth yield based on consumption of oxygen :
YX /O
O2
S S assimilation S assimilation S growth energy S maintenance

into biomass
into an
energy
extracellular
product

Yield coefficients: defined based on the amount of
consumption of another material.
For most bacteria and yeast:
Yx/s =0.4-0.6 g/g glucose
Yx/O =0.9 1.4 g/g O2
2
At the end of the batch growth period, the measured yields are
apparent as of endogenous metabolism occurring, Kd > 0,
which changes the metabolic pathways of the substrate.
M
App
e.g.
YX / S YX / S

Growth Kinetics Calculation

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What are the different phases of bacterial growth in a batch culture?

What are the different phases of bacterial growth in a batch culture?

What factors can prolong the lag phase?

What factors can prolong the lag phase?

Cell Growth Kinetics

Gross specific growth rate (1/time)

The rate of loss of cell mass due to cell death

Endogenous metabolism: during the stationary phase,

: Cell number concentration (cell number /L)

Typical growth curve for a bacterial population

Batch Growth Kinetics

Typical growth curve for a bacterial population

Typical growth curve for a bacterial population

Batch Growth Kinetics

Batch Growth Kinetics

The slope net is constant.

Typical growth curve for a bacterial population

Batch Growth Kinetics

Typical growth curve for a bacterial population

Batch Growth Kinetics

Typical growth curve for a bacterial population

Batch Growth Kinetics

Kinetic Pattern of Growth and

Growth-associated Mixed-growth-associated Non growth-associated

Batch Growth Kinetics

Batch Growth Kinetics

Batch Growth Kinetics

S S assimilation S assimilation S growth energy S maintenance

Batch Growth Kinetics

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