Kinetics in continuous

Fermentation
Batch culture:
• A fixed volume of culture medium that is continually
being altered by the metabolic activities of growing
organism and is therefore a closed system.
• In the early stage of exponential growth in batch
culture, conditions may remain relatively constant,
• In later stages when cell numbers become quite
large, drastic changes in the chemical composition of
the culture medium occur.
Continuous culture:

• An open system of contrast volume to which fresh medium

added continuously and spent culture medium is removed
continuously both at a constant rate .

• Once such a system is in equilibrium, the chemostate

volume, cell number, and nutrient status remain constant,
and the system is said to be in steady state.
• The chemostat controls both the growth rate and the
population density of the culture simultaneously .
• Two factors are important in such control:
– the dilution rate and
– the concentration of a limiting nutrient, such as a carbon or nitrogen
source.

• Growth rate and growth yield can be controlled independently

of each other,
– Growth rate - by adjusting the dilution rate and
– Growth yield - by varying the concentration of a nutrient present in a
limiting amount.

• A practical advantage to the chemostat is that a population

may be maintained in the steady state for long periods.
A steady state of the growth is regulated by the dilution Rate (D)
Hence, D = F/V
Where F – flow rate (l/h)
V – the volume (l)

The net charge in the cell concentration over time may be

expressed as :

dx/dt = growth – output or dx/dt = µx – Dx

Under steady state conditions, the cell concentration remains
constant,
Therefore

dx/dt= 0 and µx = dx and µ = D

• Continuous culture may be operated at dilution rates
below the maximum specific growth rate and so
within certain limits
• The dilution rate may be used to control the growth
rate of the culture.
• Cell growth in such a continuous culture is controlled
by the availability of the growth limiting substrate and
the system is referred to as a chemostat.
The mechanism underlying the controlling effect of the dilution
rate is expressed in the Monod equation:

µ = µ m s/(Ks +s)
At steady state, µ
=D
Therefore, D = µ mŠ (/Ks +Š)
Š = is the steady state concentration of substrate in the chemostat.

Rearrange this equation:

Š = KsD/(µm-D)

Which predicts that the substrate concentration is determined by the dilution

rate.
 Š = KsD/(µm-D)

• This occurs by the growth of the cells depleting the substrate

to a concentration that supports the growth rate equal to the
dilution rate.
• If the substrate become depleted below the level that
supports the growth rate dictated by the dilution rate, the
following will occur:

1. The growth rate of the cells will be less than the dilution rate and
they will be washed out of the vessel at a rate greater than they are
being produced resulting in a decrease in biomass concentration.
2. The substrate concentration in the vessel will rise because fewer
cells are left in the vessel to consume it.
3. The increased substrate concentration in the vessel will result in the
cells growing at a rate greater than the dilution rate and biomass
concentration will increase.
4. The steady state will be re-established.
 Therefore a chemostat is a nutrient-limited, self-balancing
culture system which may be maintained in a steady state
over a wide range of sub-maximum specific rates.

The cell concentration in a chemostat is defined by:

Ẍ= Y(SR – Š)
Where Ẍ is the steady state cell concentration.
By combining equation of steady state substrate and biomass
concentrations:
Ẍ = Y[SR – {KsD/(μm – D)}]
Therefore biomass concentration at steady state is defined by operational
variables SR and D.
 If SR is increased, Ẍ increases but Š remains the same.
 If D is increased, μ will increase (μ = D), at the new steady state would have
increased to support the elevated growth rate and less substrate will be
available to be converted to biomass resulting in a lower Ẍ
An alternative type of continuous culture to a chemostat is a
turbidostat.
Here the concentration of the cells in the vessel is kept constant by controlling
the flow of medium such that the turbidity of the culture is kept within certain
narrow limits.

 To achieve this, biomass is monitored using a photoelectric cell, signals are

sent to a pump controlling medium flow into the vessel.
 If the biomass exceeds a set point, the pump is switched on and if the
biomass falls below the set point it is switched off.
 Other biomass measurement systems include CO2 concentration or pH -
biostat.
 However, the chemostat is the more commonly used system as it has the
advantage over the biostat of not requiring complex control systems to
maintain steady state.
The kinetic characteristics of an organism are described by the numerical values
of the constants Y, µmax and Ks .

 The value of Y affects the steady-

state biomass concentration;
 The value of µmax affects the
maximum dilution rate
 The value of Ks affects the residual
substrate concentration and also the
maximum dilution rate

A hypothetical bacterium with a low Ks value for the limiting substrate, compared
with the initial limiting substrate concentration. With increasing dilution rate, the
residual substrate concentration increases only slightly until D approaches µmax when s
increases significantly. The dilution rate at which x equals zero (that is, the cells have
been washed out of the system) is termed the critical dilution rate (Dcrit) and is given by
the equation:
Dcrit = µmaxSR/(Ks + SR)
Thus, Dcret is affected by the constants, µmax and Ks., and the variable, SR;
the larger SR the closer is Dcret to µmax .
However, µmax cannot be achieved in a simple steady state chemostat because substrate limited
conditions must always prevail.
A hypothetical bacterium with a high Ks for the limiting substrate
compared with the initial limiting substrate concentration

• With increasing dilution rate, the

residual substrate concentration
increases significantly to support
the increased growth rate.
• Thus, there is a gradual increase
in s and a decrease in x as D
approaches Dcrit'
The effect of increasing the initial limiting substrate concentration
on Ẍ and Š.

As SR is increased, so Ẍ increases,
but the residual substrate
concentration is unaffected.
Also Dcrit increases slightly with
an increase in SR.

Imperfect mixing would increase in the degree of heterogeneity in the fermenter

-some organisms being subject to nutrient excess
-others under severe limitation.
This phenomenon is particularly relevant to very low dilution rate system when
the flow of the medium is likely to be very intermitted. This problem may be
overcome by the Fed-batch system.