Microbial Growth kinetics

Cells growth
• Microbial growth can be defined as an orderly increase in
cellular components, resulting in cell enlargement and
eventually leading to cell division.
• This definition is not strictly accurate as it implies that a
consequence of growth is always an increase in cell numbers.
However, under certain conditions growth can occur without
cell division, for example, when cells are synthesizing storage
compounds, e.g. glycogen or poly b-hydroxybutyrate.
• In this situation the cell numbers remain constant, but the
concentration of biomass continues to increase. This is also
true for coenocytic organisms, such as some fungi, that are
not divided into separate cells. Their growth results only in
increased size.
• The cell concentration is measured by direct and
indirect methods.
• Direct methods include measuring the cell mass
concentration and cell number density by its dry
weight, turbidity (optical density), plate counts
etc.
• Whereas, indirect methods of measuring cell
density are done by measuring the
concentration of proteins, ATP or DNA content.
 Microbial fermentations

• Microbial fermentations in liquid media can be carried out

under different operating conditions, i.e. batch growth, fed-
batch growth or continuous growth.
• Batch growth involves a closed system where all nutrients are
present at the start of the fermentation within a fixed volume.
The only further additions may be acids or bases for pH control,
or gases (e.g. aeration, if required).
• In fed-batch systems fresh medium or medium components are
fed continuously, intermittently or are added as a single
supplement and the volume of the batch increases with time.
• Continuous fermentations are open systems where fresh
medium is continuously fed into the fermentation vessel, but
the volume remains constant as spent medium and cells are
removed at the same rate.
Batch growth
• In the lag phase virtually no growth occurs and
the microbial population remains relatively
constant. Nevertheless, it is a period of intense
metabolic activity as the microbial inoculum
adapts to the new environment.
• It is usually longer if the inoculum was grown up
using a carbon source different from that in the
fresh medium, because the cells must synthesize
enzymes required to catabolize the new
substrate.
• Other factors influencing the length of the lag
phase are the age, concentration, viability and
morphology of the inoculum.
• Once the cells have adapted to their new
environment they enter the acceleration phase.
Cell division occurs with increasing frequency
until the maximum growth rate (µmax) for the
specific conditions of the batch fermentation is
reached.
• At this point exponential growth begins and
cell numbers/biomass increase at a constant
rate.
• Mathematically, this exponential growth can be
described by two methods; one is related to
biomass (x) and the other to cell numbers (N). For
cell biomass, growth can be considered as an
autocatalytic reaction.
• Therefore, the rate of growth is dependent on the
biomass concentration, i.e. catalyst, that is present
at any given time. This can be described as follows:
rate of change of biomass is dx/dt=µx eq. 1
where x = concentration of biomass (g/L),
µ = specific growth rate (per hour) and
t = time (h)
Equation 1 can also be rearranged to estimate the
specific growth rate (µ):
µ= 1/x * dx/dt eq 2
During any period of true exponential growth,
equation 1 can be integrated to provide the following
equation:
xt=xoeµt eq 3
where xt = biomass concentration after time t,
xo = biomass concentration at the start exponential
growth,
and e = base of the natural logarithm
Taking natural logarithms, loge (ln), gives
ln xt= ln xo+ µ t eq 4
• For cells in exponential phase, a plot of natural
log of biomass concentration against time, a
semilog plot, should yield a straight line with the
slope (gradient) equal to µ.
µ = (ln xt – ln xo)/t
• This equation is of the form y = c (intercept on y
axis) + mx where m= gradient, which is the
general equation for a straight-line graph.
Fig. Exponential growth of a unicellular
microorganisms with a doubling time of 20 min
(a) arithmetic plot (b) Semilog plot using
logrithms
• A second approach is to examine growth in relation to
cell number, where the number of cells at the start of
exponential growth is No.
• If we consider a hypothetical case where the microbial
cell population at this point of exponential growth is
one (No=1), we can describe binary fission as follows:
Generation time for selected bacteria