Kinetics o f B i o l o g i c a l

Processes
 Learning Outcomes

 Kinetics of cell/biomass growth

 Kinetics of product formation
 Kinetics of substrate utilization

 To propose a bioreactor system for plant,

animal cell cultures
 To select the bioreactor operation modes for
fermentation and cell growth
 What do you
understand about
growth of cells?

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Growth: basic concepts

Precursors

Anabolism = biosynthesis
Catabolism = reactions to
recover energy (often ATP)
 Growth of Cells
1. Replication and change in cell size
2. Cells grow under physical, chemical and
nutrional conditions
3. Cells extract nutrients from the medium and
convert them into biological compounds
4. Nutrients used for - energy, biosynthesis, product
formation
5. Result of nutrient utilization, microbial mass
increases with time:

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 a) Direct methods
1. Dry weight (filtration or centrifugation)
2. Optical density (OD) (light scattering, nm)

1. Dry weight 2. OD (turbidity)

• More applicable for mixed culture
application
•Using method of Volatile
Suspended Solid (VSS). Dried at
550oC for 24 hours; dry weight is
measured
• More applicable for pure culture
• Using a spectrometer
• Fast, inexpensive, simple method
•Using calibration curve to relates OD
to dry-weight
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b) Indirect methods
Measure biomolecule concentration and correlate to dry
cell mass concentration.

• Intracellular components of cells – RNA, DNA, protein, ATP

• Measurement of substrate consumption and/or product

formation during the course of growth
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• Adaptation of cells to a new environment

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• Known as late log phase
• Slowed growth due to
1. Nutrient depletion – 1 or more
2. Waste accumulation – toxic byproducts
3. Unbalanced growth and metabolism
shifts for survival 14
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• Known as resting phase

• no net growth of cell numbers or

cell mass (no cell division)

• cell growth rate = cell death rate

• secondary metabolites (products) produced

• endogenous metabolism of energy stores can result

in maintaining cell viability

• removal of inhibitory compounds will result in

further growth if additional substrate is provided 15
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Known as decline phase

Cell lysis (spillage) may occur – intracellular nutrients released into

the medium – used by living organisms during stationary phase

Growth can be re-established by transferring to fresh media

At the end of the stationary phase:

• Total nutrient depletion and toxic product
accumulation
• Death phase begin
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 Stoichiometric Coefficients for Growth

Yield coefficients, Y, are defined based on the amount of consumption of another

material

• For organisms growing aerobically on glucose,

Unit = g dry cells/g substrate consumed

Kinetics of product formation

Growth Non-growth
Associated Associated
Products Product

Mixed Growth
Associated
Product
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 Kinetics of product formation
1. Growth associated products: products appear
simultaneously with cells in culture

e.g., production of a enzyme kinetics 19

 Kinetics of product formation

2. Non growth-associated: product formation occurs during

Stationary Phase when the growth rate is zero
• Specific rate of product formation is constant

Secondary metabolites –antibiotics (e.g., penicillin) 20

 Kinetics of product formation

3. Mixed-growth associated products: products

appear during slow growth and stationary phase

Specific rate of product formation:

e.g., lactic acid fermentation, xanthan
gum, & some secondary metabolites. 21
 Growth Kinetics MODEL : Monod Equation

Monod equation is a kinetic model which

describes microbial growth as a functional
relationship between the specific growth
rate and an essential substrate
concentration

Similar to Michaelis-Menten, Langmuir-Hinshelwood (Hougen-

Watson)
• This is the most common kinetic model for cell growth.
 Growth Kinetics MODEL : Monod Equation

Similar to Michaelis-Menten, Langmuir-Hinshelwood

(Hougen-Watson)
• Assumes that a single enzyme system is responsible for the uptake of
substrate (S), and that S is limited (growth-dependent variable).
• This is the most common kinetic model for cell growth.

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 Monod equation
1 𝑑𝑋 𝜇 𝑚𝑎𝑥 𝑆
× =𝜇 =
𝑋 𝑑𝑡 𝐾𝑠 + 𝑆

Where:
X = concentration of bacteria degrading the substrate.
µ = Specific growth rate of bacteria
µmax = Maximum value of specific growth rate
S = Concentration of limiting substrate or nutrients
Ks = Saturation constant, equal to the concentration of substrate giving
growth rate of µmax
 Application and importance
If the concentration of S is
reduced, the population
growth rate will decrease. If
concentration of S increases
to a specific limit where
growth rate is maximum, then
S is no longer regarded as a
limiting factor.

When Ks = S the term S/(Ks +

S) becomes half (1/2) and the
growth rate becomes equal to
Figure 1 is a graphic representation of Monod’s equation ½ maximum rate.
(Von Sperling and De Lemos Chernicharo 2005).

(Von Sperling and De Lemos Chernicharo 2005).

 The aim of Monod was to establish that bacterial growth
rate was a function of the substrate concentration increase
to a certain level where the rate of growth becomes
constant with increased substrate concentrations.
Thank you