Module 1.2 - Growth Kinetics

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1 Module 1 – Basics of Bioprocess Technology

GROWTH CURVE KINETICS
Microbial growth is the cell division, of a parent cell into two daughter cells, in a process called binary fission. The rate of
growth of a cell population vary with time after the inoculation of cells into culture medium. After a small population of
microbial has been inoculated into fresh culture medium, the population of living cells is counted at regular intervals. The
curve may be drawn by comparing the total number of living bacteria present in the population over a period of time. The
population growth curve has four major phases:
1. Lag Phase
When an organism is inoculated in fresh culture medium, the population does not double immediately. The total number
of living cells remains fairly constant for a long period. This is known as lag phase. The organisms adjust to its new
environment and starts to synthesize molecules that are required for its survival i.e. enzymes, coenzymes etc. In short, the
organisms are metabolizing and growing, but there is a lag in cell division. The duration of lag phase varies with the
conditions and species. The lag phase follows Zero Order Kinetics i.e. they are independent of substrate present as they
are trying to adjust to the environment.
2. Log Phase
Cells in the log phase are constantly reproducing at the exponential rate and demonstrate most of the typical features of
the species and the curve shows a straight line. Also, the log phase cultures are highly sensitive to various physical and
chemical antimicrobial agents. The log phase follows First Order Kinetics of Exponential Growth i.e. they are dependent
on the substrate for growth. The log phase is the time when the cells are metabolically most active, and in industrial
production, this is the peak period of activity and efficiency. So, this phase is most useful for fermentation industries.
3. Stationary Phase
The decreasing essential nutrients and accumulating toxic waste products discourage or reduce the growth rate of the log
phase. The factor that inhibits the microbial growth is called growth limiting factor. The cells in stationary phase have a
chemical composition different from the cells in exponential phase and the cells are small since division continues after
increase in mass has stopped. They are more resistant to adverse physical and chemical agents. This phase again follows
Zero Order Kinetics i.e. they become independent of the substrate present in the environment. The no of cells produced in
this phase is equal to no of cells dying.
4. Death Phase
The death phase of the growth curve is basically the reverse of log phase. So, it follows the First Order Decay. Cells die
and are destroyed by lysis at an exponential rate. Their death and lysis are primarily the result of an ever increasing
amounts of acids and other harmful wastes in the environment and depletion of essential nutrients from culture medium.
Under ideal conditions for growth, when a batch growth or batch culture is carried out, it can be observed experimentally
that the quantity of biomass, and therefore also the concentration, increases exponentially with respect to time. This
phenomenon can be explained by the fact that all cells have the same probability to multiply.
Overall rate of biomass formation is proportional to the biomass itself. This leads to an autocatalytic reaction, which is
described by a first order rate expression.
dX
r x= =μX (1)
dt
Where
rx = Rate of cell growth,
X = Cell concentration
µ = Kinetic Growth Constant (1/s) or Specific Growth Rate.
dX
= Rate of change of cell concentration with respect to time.
dt
The Microbial Growth Curve Equation

Let ‘X’ be the no of cells at time ‘t’. The cells, as found experimentally, have
been found to increase by a factor of 2 as a single parent cell divides into 2
daughter cells and so, the value comes in existence. So, the no of cells (X) after a
particular period of time is:
X = 2Z (2)
But this is applicable only when the initial no. of cells is equal to 1. When it is more than one, which is always the
practical case, the initial no of cells has to be taken into consideration and thus comes a new constant in the equation (2)-
X = X0. 2z (3)
z
Where, 2 = Increase in the no of microbes
Z = Generation number
X0 = No of cells at‘t=0’.
Taking natural log on both sides,

ln X = ln X0 + z.ln2 (4)
Which gives
ln X−ln X 0
z=
¿2
(5)
But also,
t
z= (6)
T
Where, t = Time period of exponential growth
T = Mean generation time or Doubling time
Therefore, taking equation 5 and 6 into consideration, the following equation rises-
t ln X−ln X 0
=
T ln 2
(7)
But, as per the graph obtained experimentally, it was found that the equation does not consider the time taken by the cells
while undergoing lag phase. So equation 7 was modified as-
t−L ¿ X−¿ X 0
=
T ¿2
(8)
Where, L = lag phase time of the overall biomass production time.
Equation no 8 gives the mathematical interpretation of the sinusoidal graph obtained from a microbial growth system.
Relation between Mean Generation time or Doubling time and Specific Growth Rate
From equation 6, it is understood that the generation no is related to the Doubling time as well as time period of the
microbial growth batch. But also, it is found that the reciprocal of Doubling time (T) gives what is called the Doubling
Rate (v)-
1
=v (9)
T
Also, with respect to exponential function,
2 = e ln2 (10)
Taking Eq. 10 into Eq. 3,
X = X 0 (eln2) Z = X0 (eln2) t/T = X0 (eln2/T) t = X0. e µt (11)
Where, µ = Specific Growth rate of the cell
Now, relating the newly found Specific Growth Rate ‘µ’ with Doubling Time T -
¿2 ¿ 2 0.693
µ= T= = (12)
T μ μ
Microbial death rate equation w.r.t rate of growth and rate of death
In many processes cells begin to die (after t s), because of lack of nutrients, toxic effects or cell aging. This process can
typically be described by a first order decay. But since there is some growth still happening that is sustaining the no of
cells, the mathematical equation for all cells at death phase is
dN
=¿ (µ-kd)X (13)
dt
The equation can be used to describe the whole microbial growth curve –
 When the cells are in lag phase, both µ and kd are insignificant in value but are present.
 When the cells start with acceleration phase, k d is still insignificant but µ suddenly increases drastically.
 When cells are at exponential phase, µ is very large as compared to k d and hence only µ is considered in the
equation.
 As the stationary phase starts, rate of death of cells comes approximately near to the rate of microbial cell growth.
 As the death phase starts, µ becomes significantly less as compared to k d and hence the equation becomes
r =−k d X (14)
Where
r = Death Rate and
kd = Specific Death Rate, with the same dimensions as the specific growth rate.

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1 Module 1 – Basics of Bioprocess Technology

The Microbial Growth Curve Equation

Taking natural log on both sides,

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