Professional Documents
Culture Documents
KINETICS OF GROWTH
IN BATCH AND
CONTINOUS CULTURE
1 dX
net
X dt
Cell growth
The net specific growth rate is the difference between
a gross specific growth rate (g, h-1) and the rate of
loss of cell mass due to cell death (kd, h-1):
net g kd
Microbial growth can also be described in terms of
cell number, N:
1 dN
R
N dt
where R is the net specific replication rate (h-1)
Batch Growth Determining Cell
Number Density
Hemocytometer
Direct microscopic count
Counts all cells present (viable and non-viable)
Immediate result
Agar plates
Counts only living cells
Delayed result
Assumption: each viable cell will yield 1 colony
Results expressed in CFUs(colony-forming units)
Particle counters
Counts all cells present (viable and non-viable)
Suitable for discrete cells in a particulate-free medium
Can distinguish between cells of different sizes
Hemocytometer
Viable Cell Count
Coulter Particle Counter
Determining Cell Mass Concentration Direct
Methods
Growth Phases
1. Lag
2. Exponential
3. Deceleration
4. Stationary
5. Death/Decline
Lag Phase
Occurs immediately after inoculation and is a
period of adaptation for the cells to their new
environment
New enzymes are synthesized, synthesis of other
enzymes is repressed
Intracellular machinery adapts to the new conditions
May be a slight increase in cell mass and volume, but
no increase in cell number
The lag phase can be shortened by high inoculum
volume, good inoculum condition (high % of living cells),
age of inoculum, nutrient-rich medium
Influence of [Mg2+] on Lag Phase Duration in E.
aerogenes Culture
E. aerogenes requires Mg2+ to
activate the enzyme
phosphatase, which is
required for energy
generation by the organism
The concentration of Mg2+ in
the medium is indirectly
proportional to the duration
of the lag phase
Exponential Growth Phase
In this phase, the cells have adjusted to their new
environment
At this point the cells 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 first order exponential growth rate expression is:
dX
net X where X X 0 at t 0
dt
X
ln nett or X X 0 e nett
X0
Exponential Growth Phase (contd)
An important parameter in the exponential phase is
the doubling time (time required to double the
microbial mass)
A graph of ln X versus t produces a straight line on a
semi-logarithmic plot:
ln 2 0.693
d
max max
The doubling time based on cell number is expressed
as: ln 2
d
'
R
Exponential Growth Phase (cont)
t
Deceleration Phase
Very short phase, during which growth decelerates
due to either:
Depletion of one or more essential nutrients, or,
The accumulation of toxic by-products of growth (e.g.
Ethanol in yeast fermentations)
Period of unbalanced growth: td=td
Cells undergo internal restructuring to increase their
chances of survival
Followed quickly by the Stationary Phase
Stationary Phase
Starts at the end of the Deceleration Phase, when the
net growth rate is zero (no cell division, or growth rate
is equal to death rate)
Cells are still metabolically active, and can produce
secondary metabolites
Primary metabolites are growth-related products, while
secondary metabolites are non-growth-related
Many antibiotics and some hormones are produced as
secondary metabolites
Secondary metabolites are produced as a result of
metabolite deregulation
Stationary Phase (contd)
During this phase, one or more of the following
phenomena may occur:
Total cell mass concentration may stay constant, but the
number of viable cells may decrease
Cell lysis may occur, and viable cell mass may drop. A
second growth phase may occur as cells grow on lysis
products from the dead cells (cryptic growth)
Cells may not be growing, but may have active
metabolism to produce secondary metabolites
Stationary Phase (contd)
During the stationary phase, the cell catabolizes cellular
reserves for new building blocks and for energy-
producing monomers
This is called endogenous metabolism
The cell must expend maintenance energy in order to stay
alive
The equation that describes the conversion of cellular mass into
energy, or the loss of cell mass due to lysis during the
stationary phase is:
dX kd t
kd t or X X SOe
dt
Death Phase
The death or decline phase is characterized by the
expression:
dN k d' t
kd t or N N S e
'
dt
Where Ns is the concentration of cells at the end of
the stationary phase, and is the first-order death-rate
constant
A plot of ln N versus t yields a line of slope kd
Death Phase
1. Cell lysis (spillage) may occur
2. Rate of cell decline is first-order
m
dS / dt m
X
However, during the Stationary Phase, where little external
substrate is available, endogenous metabolism of biomass
components is used for maintenance energy
Maintenance energy is the energy required to repair
damaged cellular components, to transfer nutrients and
products in and out of cells, for motility, and to adjust the
osmolarity of the cells interior volume
Microbial Products
Microbial products can be classified into three major
categories
Growth-associated products
Non-growth-associated products
Mixed-growth-associated products
Growth-associated products
These products are produced simultaneously with microbial
growth
Specific rate of product formation is proportional to the
specific growth rate, g
Note that g is not equal to net, the net specific growth rate,
when endogenous metabolism is occurring
Growth-Associated Products
The rate expression for product formation in
growth-associated production is:
1 dP
qp YP / X g
X dt
Where qp is the rate of product formation (h-1)
The production of a constitutive (continuously
produced, as opposed to inducible) enzyme is an
example of a growth-associated product
Non-Growth-Associated Products
Non-growth-associated product formation takes
place during the Stationary Phase, when the growth
rate is zero
Specific rate of product formation is constant:
q p constant
Many secondary metabolites, such as most
antibiotics (e.g. penicillin), are non-growth-
associated products
Mixed-Growth-Associated Products
Mixed-growth-associated product formation takes place during
the Deceleration (slow growth) and Stationary Phases
The specific rate of product formation is given by the
Luedeking-Piret equation:
q p g
If = 0, the product is completely non-growth associated; If =
0, the product is completely growth-associated
Examples: lactic acid fermentation, production of xanthan gum,
some secondary metabolites
Product Yield Coefficients (cont)
Legend:
() Growth on
rich, complex
medium
() Growth on
glucose-mineral
salts medium
Effect of pH on Cell Growth
pH affects the activity of enzymes, and therefore
the microbial growth rate
Acceptable pHs for growth are typically within 1 or
bacteria (most) pH = 3 to 8
yeast pH = 3 to 6
plants pH = 5 to 6