OXYGEN TRANSFER

IN BIOPROCESSES

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 Topic Outline
1. Define oxygen transfer
2. Importance of oxygen transfer in biological systems
3. Oxygen transfer steps
4. Two film theory applications in bioprocesses
5. Oxygen transfer limitations in bioprocesses
6. Oxygen transfer measurement methods

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 Importance of Aeration
• Oxygen supply to cells
• Removal of gaseous products (CO2,N2)
• Low solubility of oxygen in water
 subsequent supply is required
• Solubility of oxygen depends on temperature
and pressure

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Factors influencing oxygen supply

(a) Process factors

(b) Transfer through an interface (KLa)
(c) Determination of KLa;
(d) Factors affecting bubble size
(e) Gas hold-up
(f) Economics of oxygen transfer

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 Oxygen supply
• Oxygen is normally supplied to microbial cultures in the form of air, i.e.
cheapest source

• Provision of oxygen varies with the scale of the process, compare lab
and industrial process design

• Oxygen mass balance depends on rate at which O 2 can be delivered to

the biological system (OTR) and the rate at which it is utilised (COD)

• OTR can be increased by elevating the pressure, enriching the inlet air
with O2 and increasing agitation and aeration

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 Principles of Oxygen Supply
• Movement of oxygen in bioprocessing is as a
result of Molecular Diffusion
• This is the movement of components of
molecules in a mixture from a region of higher
concentration to a region of lower
concentration.
• By constantly supplying material in the high
concentration region we ensure its continuous
supply.
 Principles of Oxygen Supply
• The rule of movement or flux is given by Fick’s
Law.

Flux =
 Importance of Diffusion
1. Mixing – Bulk mixing of reactor contents
results in formation of eddies, however fluid
flow within the smaller eddies is largely laminar,
diffusion can therefore ensure that at a
molecular level there is adequate mixing
 Importance of Diffusion
2. Bioprocessing systems are characterized by
Clumps,flocs and films are of cells, immobilised
enzymes etc. As the reaction proceeds diffusion
ensures that product moves away from site of
reaction and reactant eg Oxygen moves to site
of reaction. Diffusion allows for intra-particle
mass transfer.
 Importance of Diffusion
3. Diffusion allows for mass transfer across
phase boundaries. The oxygen can be
transferred from the gas bubbles to
fermentation broths.
When different phases come into contact, the
fluid velocity near the phase interface is
significantly decreased and diffusion becomes
crucial at that stage.
 How to improve the oxygen transport?
• Increase of the O2-solubility
– Pressure increase from 100 to 200 kPa dC
• Increase the O2-content in the air
 k L a  (C *  C )
dt
– enrichment of the aeration with O2
– Use of pure O2
• Change in the phase boundary (gas/liquid)
– size and distribution of the gas bubbles
– contact time between the gaseous phase and the liquid phase
• Viscosity of the nutrient solution
– viscosity reduction  increases the relative velocity of the gas bubbles  thinner liquid
film  higher kLa-value
• Insertion of surface-active substances (controversial)

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Oxygen transfer steps: Gas exchange between gas bubble
and cell
interface interface gas/cell
gas/liquid

O2-concentration liquid cell

C*

Gas Bubble
cell
GB gas bubble
GF gas film
FF liquid film
C* O2-Concentration in the gas bubble
C O2-Concentration in the liquid

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Oxygen Path
From A Bubble To An Immobilized Cell System

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 Two film Theory
• It postulates that near the interface of two
phases there exists a stagnant film of
thickness (δ) .
• This stagnant film is hypothetical since we
really don't know the details of the velocity
profile near the interface.
• In this film transport is governed essentially by
molecular diffusion.
 The Two film Theory

pG pGi
pGi = H CGi
•
Bulk Gas Gas Film Liquid Film Bulk Liquid
• CLi

CL
(1/k1) (1/k2) (1/k3) (1/k4)

Gas phase resistance Liquid phase resistance

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 Two Film Theory……….2
dC/dt = kg (Pg - Pi) = KL (Ci - CL)
(Driving force)(Resistance)

Note kg = 1/k1 + 1/k2, KL = 1/k3 + 1/k4

Use conc rather than partial pressure (measure)

 dC/dt = KL (Csat - CL) ......assume that Csat substitutes for Ci (measure)

This is per unit interface!

Overall then dC/dt = KLa( Csat - CL)

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 Oxygen Path
From A Bubble To An Immobilized Cell System

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 The oxygen transfer process
Step 1 - Diffusion through the bubble to the gas-liquid
interface
Step 2 - Diffusion across the gas-liquid interface
Step 3 - Diffusion through the bubble boundary layer
Step 4 - Movement through the bulk liquid by forced
convection and diffusion
Step 5-9: Movement through the floc
Step 5 - movement through the boundary layer surrounding
the microbial slime
Step 6 - entry into the slime
Step 7 - movement through the slime
Step 8 - movement across the cell membrane
Step 9 - reaction
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 Kinetics of Mass Transfer
• The rate of mass transfer is directly
proportional to the driving force for transfer
and the area available for the transfer process
to take place.
• The coefficient of proportionality is the mass
transfer coefficient.
•
 Gas-Liquid Mass Transfer
The rate of mass transfer of component A
through the gas boundary layer is:

where
CAG = concentration of A in the bulk of the gas
CAGi is concentration of A at the gas-liquid
interface.
 Gas-Liquid Mass Transfer
and the rate of mass transfer of component A
through the liquid boundary layer is:

where
CAL = concentration of A in the bulk of the liquid
CALi is concentration of A at the gas-liquid
interface.
 Gas-Liquid Mass Transfer
• assuming that equilibrium will be attained at
the interface there is steady state flux at the
interface.
• At steady state there is no accumulation of
component A at the interface or anywhere
else in the system.
• Therefore: NAL = - NAG
 Gas-Liquid Mass Transfer
• CAGi and CALi are equilibrium concentrations
and can be related.

• Therefore
 Gas-Liquid Mass Transfer
And
 Gas-Liquid Mass Transfer
• Unfortunately, concentrations at the interface
cannot be measured so overall mass transfer
coefficients are defined.
• These coefficients are based on the difference
between the bulk concentration in one phase
and the concentration that would be in
equilibrium with the bulk concentration in the
other phase.
 Gas-Liquid Mass Transfer
• Define the overall mass transfer coefficient as:

• and

• These equations can be viewed as "resistance" expressions

where 1/Kg or 1/KL represent total resistance to mass transfer
based on gas or liquid phase concentration, respectively
• the total resistance to transfer is made up of three series
resistances: liquid film, interface and gas film.
• Mass transfer coefficients are measured experimentally or
estimated using empirical correlations from literature
 Gas-Liquid Mass Transfer
Therefore

And
 Gas-Liquid Mass Transfer
• Given CAL* = liquid phase concentration that
would be in equilibrium with the bulk gas
concentration. = CAG /m
• CAg * = gas phase concentration that would be
in equilibrium with the bulk liquid
concentration. = mCAL
 Gas-Liquid Mass Transfer

And

• In general, highly soluble gases have transfer rates

controlled by gas film and vice versa.
• For example, oxygen (slightly soluble) transfer is
usually controlled by liquid film.
• Ammonia (highly soluble) transfer is usually
controlled by gas phase film.
Balance between Oxygen Supply and Oxygen
Demand
• Cells in aerobic culture take up oxygen from broth.
• The rate of oxygen transfer from gas to liquid is
therefore of prime important, especially at high
cell densities when cell growth is likely to be
limited by availability of oxygen.
• Oxygen is poorly soluble in water the limited
amount of oxygen is quickly consumed in aerobic
cultures and must be constantly replaced by
sparging.
Balance between Oxygen Supply and Oxygen
Demand
• The rate at which oxygen is consumed by cells
in fermenters determines the rate at which it
must be transferred from gas to broth.
• The oxygen demand is influenced by:
• cell species
• culture growth phase
• nature of the carbon source in the medium.
Balance between Oxygen Supply and Oxygen
Demand
• The rate of oxygen consumption per cell,
known as the specific oxygen uptake rate.
• If QO is the oxygen uptake rate per volume of
broth and qO is the specific oxygen uptake
rate:
 Critical Dissolved Oxygen
• Dissolved oxygen is an important substrate in
aerobic fermentations.
• Since oxygen is sparingly soluble in water, it
may be the growth-limiting substrate in these
fermentations.
• For bacteria and yeast cultures, the critical
oxygen concentration is about 10% to 50% of
the saturated DO (dissolved oxygen
concentration).
 Critical Dissolved Oxygen
• When designing a bioreactor one must consider and ensure that O2 uptake
rate of the culture does not exceed the oxygen transfer rate of the
fermenter

• Uptake rate =
X = Biomass
• = supply rate
• The dissolved oxygen concentration must be above a certain level known as
the critical oxygen concentration. Ccrit which depends on the organism.
 Critical Dissolved Oxygen
• At steady state where there in accumulation
of oxygen at any location in the reactor, the
uptake rate must be equal to supply rate
• =
 Critical Dissolved Oxygen
• To evaluate the maximum cell concentration that can
be supported by a fermenter, consider the point
where we have the maximum rate of mass transfer.
• This is when the concentration driving force is at its
maximum ie CAL is zero (this is a point where all the
oxygen in the system is in equilibrium with gas
phase)
 Critical Dissolved Oxygen
• A fermentation process will have a max KLA dictated by operating conditions
thus it is the demand that often has to be adjusted.

Achieved by:
• Control of biomass conc
• Control of specific O2 uptake rate
• Combination of both
• Another important parameter is the minimum kL a required to maintain
CAL > Ccrit in the fermenter. This can also be determined as: