STOICHIOMETRY OF
MICROBIAL GROWTH AND
PRODUCT FORMATION
• BATCH GROWTH
– Growth Cycle
MARY ROSE F. PERSINCULA
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
Growth Kinetics: 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 (1/time): net
1 dX
X dt
t: the time
Growth Kinetics
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
net
: during the stationary phase, the cell catabolizes
cellular reserves for new building blocks and
for energy-producing monomers.
Growth Kinetics
Net specific replication rate (1/time):
1 dN
R
N dt
R R
' kd
R
N : Cell number concentration (cell number /L)
' : Gross specific replication rate (1/time)
R
kd : The rate of cell death (1/time)
Growth Kinetics
- Quantifying cell concentration:
- direct: no suspended solid and interference
compounds.
packed cell volume – preferred
dry weight, optical density (turbidity) (600-700nm
Wave Length)
cell or colony count :
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: culture medium contains more than
one carbon source
Diauxic growth
Typical growth curve for a bacterial population
Typical growth curve for a bacterial population
Batch Growth Kinetics
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.
Batch Growth Kinetics
Exponential growth phase
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 μnet 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.
Typical growth curve for a bacterial population
Batch Growth Kinetics
Exponential growth phase
net R μm
μm 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
Typical growth curve for a bacterial population
Batch Growth Kinetics
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
Typical growth curve for a bacterial population
Batch Growth Kinetics
Stationary Phase:
With the exhaustion of nutrients (S≈0) 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.
metabolites are growth-related: ethanol by S.
cerevisae.
metabolites are non-growth-related:
antibiotics, pigments.
Kinetic Pattern of Growth and
Product Formation
Growth-associated Mixed-growth-associated Non growth-associated
Batch Growth Kinetics
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
k d X , k d is the rate constant for endogenous metabolism.
dt
Batch Growth Kinetics
Death Phase:
The living organism population decreases with time,
due to a lack of nutrients and toxic metabolic by-
products.
The rate of death usually follows:
dN '
kd N
dt
'
k d is the first - order death rate constant.
