Microbial Growth Kinetics

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X is cell mass concentration (g/l),

t is time (h),
µnet is net specific growth rate (h-1)
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Doubling time of cell mass - The time required to

double the microbial mass
Effect of substrate concentration

• Structured and segregated,

• Structured and nonsegregated,
• Unstructured and segregated, and
• Unstructured and nonsegregated

S (substrate) - is growth-rate limiting (i.e., an increase

in S influences growth rate, while changes in other
nutrient concentrations have no effect). Monod equation:

µmax is the maximum specific growth rate

The constant Ks is known as the saturation constant and is equal to the concentration of the rate-limiting
substrate when the specific rate of growth is equal to one-half of the maximum.
Ks = S when µg = ½ µmax. µg =µmax when S > > Ks
To better describe growth kinetics, lets define some stoichiometrically related parameters.
Yield coefficients are defined based on the amount of consumption of another material.

Growth yield

Y x/s= = - Y P/s= = -

At the end of the batch growth period, we have an apparent growth yield (or observed growth yield). Because
culture conditions can alter patterns of substrate utilization. For example, with a compound (such as glucose)
that is both a carbon and energy source, substrate may be consumed as:
 Microbial products can be classified in three major categories

• Growth-associated products - produced simultaneously with microbial growth. The specific rate of product
formation (qp) is proportional to the specific rate of growth, µnet.
The production of a constitutive enzyme, ethanol are examples of a growth-associated product.
qp = µnet =
dp/dx µnet = dp/dt dt/dx µnet = dp/dt 1/x

• Nongrowth-associated product formation takes place during the stationary phase when the growth rate
is zero. The specific rate of product formation is constant.
e.g. Many secondary metabolites, such as antibiotics (for example, penicillin)

qp = = constant
• Mixed-growth-associated product formation takes place during the slow growth and stationary phases. In
this case, the specific rate of product formation is given by the following equation
qp = α µnet + Luedeking–Piret equation
Lactic acid fermentation, xanthan gum, and some secondary metabolites
Models with growth inhibitors

• At high concentrations of substrate or product and in the presence of inhibitory substances in the medium, growth
becomes inhibited, and growth rate depends on inhibitor concentration.

Substrate inhibition:
• At high substrate concentrations, microbial growth rate is inhibited by the substrate.
• Substrate inhibition of growth may be competitive or noncompetitive.

KI is the inhibition constant

• Under single-substrate inhibiting conditions, the Monod equation is no longer suitable, and the most common Monod
derivative is in the form of the Haldane equation.
• Reasons for substrate inhibition in bioreactor cell growth includes osmotic issues, viscosity, or inefficient oxygen
transport due to overly concentrated substrate in the bioreactor medium
• Substrates that are known to cause inhibition include glucose, NaCl, and phenols, among others.
• Substrate inhibition is also a concern in wastewater treatment, where one of the most studied biodegradation
substrates are the toxic phenols. Due to their toxicity, there is a large interest in bioremediation of phenols, and it is well
known that phenol inhibition can be modeled by the following Haldane equation:

(Haldane equation for single-substrate inhibition of cell growth)

• Substrate inhibition may be alleviated by slow, intermittent addition of the substrate to the growth medium.
• Competitive and non-competitive inhibitors can be told
apart by how they affect activity at different substrate
concentrations.
• If an inhibitor is competitive, it will decrease reaction rate
when there's not much substrate, but can be "out-
competed" by adding lots of substrate.
• If an inhibitor is noncompetitive, the reaction will never
reach its normal maximum rate even with a lot of
substrate. This is because the molecules with the
noncompetitive inhibitor bound are "poisoned" and can't
do their job, regardless of how much substrate is available.
Product inhibition:
• High concentrations of product can be inhibitory for microbial growth.
• Product inhibition may be competitive or noncompetitive.

• Ethanol fermentation from glucose by yeasts is a good example of noncompetitive product inhibition, and ethanol is
the inhibitor at concentrations above about 5%
Inhibition by toxic compounds:
• The following rate expressions are used for competitive, noncompetitive, and uncompetitive inhibition of growth in
analogy to enzyme inhibition.
CELLS GROWTH IN CONTINUOUS CULTURE

In batch culture - Growth, product formation, and substrate utilization terminate after a certain time interval,

We have already talked

about it in class, hope you
Everything fixed. So, bacteria keep consuming food, remember
but there is not constant supply of food.
Continuous In
Continuous Out

In continuous culture - fresh nutrient medium is

continually supplied to a well-stirred culture, and
products and cells are simultaneously withdrawn

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CELLS GROWTH IN CONTINUOUS CULTURE

• Fresh nutrient medium is continually supplied to a well-stirred culture, and products and cells are
simultaneously withdrawn

• Growth and product formation can be maintained for prolonged periods in continuous culture

• After a certain period of time, the system usually reaches a steady state where cell, product, and substrate
concentrations remain constant.

• Continuous culture provides constant environmental conditions for growth and product formation and
supplies uniform-quality product.

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The Ideal Chemostat - The primary types of continuous cultivation devices
• An ideal chemostat is the same as a perfectly mixed Continuous Stirred-Tank Reactor (CSTR).

• Fresh sterile medium is fed to the completely mixed and aerated reactor, and cell suspension is removed at the
same rate.

• Liquid volume in the reactor is kept constant.

A material balance on the cell concentration around the chemostat yields:

F - the flow rate of nutrient solution (l/h),

VR - is the culture volume (l) (assumed constant),
X - the cell concentration (g/l),
µg and kd - growth and endogenous (or death) rate constants,
respectively (h-1)
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 under steady-state conditions, the cell
concentration remains constant or vice-versa,
when dX/dt = 0, then reactor reach steady state
and the change in biomass concentration (X)
So, now if you rearranged this to over time (t) is zero i.e. X remain constant

dX/dt = X - kd X

Then,

Usually, the feed media are sterile (isn’t it?, that’s the 1st step if you remember from your lab, we sterilize the media
first by autoclaving), X0 = 0,

if the endogenous metabolism or death rate is negligible compared to the growth rate ( kd << mg or kd= 0) and if the system is
at steady state (dX/dt = 0), then

=D

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 =D So, 2 importants can be derived from this eqn

• In a chemostat, cells are removed at a rate equal to their growth rate, and the growth rate of cells is equal to the
dilution rate.

• This property allows the investigator to manipulate growth rate as an independent parameter and makes the
chemostat a powerful experimental tool

Now recall your memory, what was ? Remember the Monod eqn?

where S is the steady-state limiting substrate concentration (g/l).

If D is set at a value greater than mm, the culture cannot reproduce quickly enough to maintain itself and is washed out
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 Rate of change in Biomass concentration = Rate of Production – Rate of removal

dX/dt = mX - DX

Steady state Dilution rate decreased Dilution rate increased

D S D S
dX/dt = 0
m m
m = D