BIOCHEMICAL ENGINEERING

Transport Phenomena in Bioprocess Systems:

Gas-liquid mass transfer in bioreactors

Engr. A. Corpuz 09/2021

Chemical Engineering
Cagayan State University – Carig Campus, Tuguegarao City
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Supplying oxygen to aerobic cell is a significant
challenge
⚫ The problem：oxygen is poorly soluble in water the
solubility of oxygen in pure water is 8 mg/L at 4oC
(sucrose is soluble to 600 g/L)
⚫ The solubility of oxygen decreases as with increasing
temperature and concentration of solutes in the solution
⚫ Due to the influence of the culture ingredients, the
maximal oxygen content is actually lower than it would
be in pure water.
⚫ The solubility of gases follows Henry's Law in the gas
pressure range over which fermenters are operated.
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 Henry's Law
⚫ Describes the solubility of O2 in nutrient
solution in relation to the O2 partial pressure in
the gas phase C* =
Po
H
⚫ C* is the oxygen saturation concentration of
the nutrient solution, Po is the partial pressure
of the gas in the gas phase and H is Henry's
constant, which is specific for the gas and the
liquid phase
⚫ Aeration with air 9 mg O2/L dissolves in water,
with pure oxygen 43 mg O2/L. 3
 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 liquid film surrounding
interface
Step 4 - Movement through the bulk liquid by forced
convection and diffusion
Step 5-9: Movement through the floc
Step 5 - movement through the liquid layer surrounding the
microbial slime
Step 6 - entry into the slime
Step 7 - movement through the slime
Step 8 - movement across the cell membrane 4

Step 9 - reaction
Oxygen Path
From A Bubble To An Immobilized Cell System

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Step 1
Diffusion through the bubble to the gas-liquid interface
▪ Gas molecules move quickly
▪ They are evenly distributed throughout the bubble.

O2

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⚫ Step 2 - Diffusion across the gas-liquid interface
This step will be very rapid if the concentration of oxygen in
the bubble is high. High oxygen concentrations in the bubble (as
measured in terms of partial pressure) will push the oxygen
molecules across the interface, into the boundary layer.
If the medium is rich in CO2 , then the carbon dioxide will be
pushed into the bubble.
If the bubble contains a low concentration of oxygen,
then the rate of oxygen transfer
out of the bubble will be slow
or even zero
O2

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CO2
Step 3- Diffusion through stagnant liquid film
The movement of solutes through the film is slow. Solutes
move through the liquid by diffusion.
The movement of the molecule will be driven by the
concentration gradient across the boundary layer.

Factors which may affect the rate of diffusion of oxygen

through the liquid film
⚫ Temperature
⚫ Concentration of oxygen in the bulk liquid
⚫ Saturation concentration of oxygen in the liquid
⚫ Concentration of oxygen in the bubble

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Step 4 Movement through the bulk liquid by forced
convection and diffusion
The rate of movement of an oxygen molecule through the
bulk liquid is dependent on
⚫ the degree of mixing (relative to the volume of the reactor)
⚫ viscosity of the medium

O2

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⚫ Step 5-9: Movement through the floc
complete the journey of the oxygen molecule
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
⚫ Steps 5 and 7 are slow processes.

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Step 3
The interface oxygen transfer equation

N A = kL  a  (C * −CL ) = OTR
NA = Volume-dependent mass transfer (mM O2 / h)
kL = Transfer coefficient at the phase boundary
a = Specific exchange surface
kLa = Volumetric oxygen transfer coefficient (h-1)
C* = Saturation value of the dissolved gas in the phase boundary
CL = Concentration of the dissolved gas (mM)
OTR = O2 Transfer Rate (mM O2/h) 11
Molecular Diffusion
The movement of component molecules in a mixture
under the influence of a concentration difference in
the system. Diffusion of molecules occurs in the
direction required to destroy the concentration
gradient.

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Fick’s Law of diffusion

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 Mass-Transfer Coefficient (kL & kG)

where
• dCL / dt is the oxygen transfer rate, mM / h
• dCL is the dissolved concentration of the solute in
the bulk liquid or fermentation broth
• k is the mass transfer coefficient for the solute
through the boundary layer
• A is the total interfacial area and
• C* is the concentration of the solute in the boundary
layer.

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 Mass-Transfer Coefficient (kL & kG)

where
• NG mass flux from the gas phase to the interface
• NL liquid-side mass flux, kg / h. m2
• qG rate of mass transfer in liquid phase, kg / h
• kG is the individual mass-transfer coefficient for the
gas phase, m/s
• A is the interfacial area , m2
• CGi is the gas-side concentration in the interface
• Cli is the liquid-side conc in the interface, kg / m3

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Concentration profile near a gas-liquid interface

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Mass-Transfer Coefficient (kL & kG)
⚫ Since the amount of solute transferred from the gas
phase to the interface must equal that from the
interface to the liquid phase,
NG =NL (1)
⚫ Substitution of NG and NL into Eq. (1) gives

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 Mass-Transfer Coefficient (kL & kG)
⚫ It is hard to determine the mass-transfer coefficient
because the interfacial concentrations, CLi or CGi
cannot be measured
⚫ To define the overall mass-transfer coefficient as
follows :

where
⚫ CG* is the gas-side concentration which would be in equilibrium with the

existing liquid phase concentration

⚫ CL* is the liquid-side concentration which would be in equilibrium with the
existing gas-phase concentration. 18
 Mass-Transfer Coefficient (kL & kG)
Gas-phase conc

L-phase conc
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⚫ The equilibrium curve explaining the meaning of CG* and CL*
 Mass-Transfer Coefficient (kL & kG)
⚫ Example 9.2 Derive the relationship between the overall mass-transfer
coefficient for liquid phase KL and the individual mass-transfer
coefficients, kL and kG. How can this relationship be simplified for
sparingly soluble gases?
For sparingly soluble gases, the slope of the
equilibrium curve is very steep; therefore, M
is much greater than 1:
KL≈kL

Similarly, for the gas-phase mass-transfer

coefficient:
KG≈kG
▪ since,

Since

Where, M is slope of the line connecting

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(CLI, CLI) and (CL*, CG)
 Mechanism of Mass Transfer
Film Theory

The two-film theory is a useful model for mass transfer

between phases. Mass transfer of solute from one phase to
another involves transport from the bulk of one phase to
the phase boundary or interface, and then from the
interface to the bulk of the second phase.

The film theory is based on the idea that a fluid film or

mass-transfer boundary layer forms wherever there is
contact between two phases.
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Exercise
9.1 Derive the relationship between the overall mass transfer
coefficient for gas phase KG and the individual mass-
transfer coefficients, kL and kG. How can this relationship be
simplified for sparingly soluble gases?

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According to the film theory, turbulence in each fluid
dies out at the phase boundary. A thin film of relatively
stagnant fluid exists on either side of the interface;
mass transfer through this film is effected solely by
molecular diffusion. The concentration of A changes
near the interface.
Most of the resistance to mass transfer resides in the
liquid films rather than in the bulk liquid.
It is generally assumed that there is negligible resistance
to transport at the interface itself; this is equivalent to
assuming that the phases are in equilibrium at the plane
of Mass Transfer contact.
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The difference between CA1i and CA2i at the interface
accounts for the possibility that, at equilibrium, A may be
more soluble in one phase than in the other.
For example, if A were acetic acid in contact at the
interface with both water and chloroform, the
equilibrium concentration in water would be greater than
that in chloroform by a factor of between 5 and 10.

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 Correlation for Mass-Transfer
Coefficient
⚫ Mass-transfer coefficient is a function of physical
properties and vessel geometry
⚫ Because of the complexity of hydrodynamics in
multiphase mixing, it is difficult, if not impossible, to
derive a useful correlation based on a purely
theoretical basis
⚫ It is common to obtain an empirical correlation for
the mass-transfer coefficient by fitting experimental
data. The correlations are usually expressed by
dimensionless groups since they are dimensionally
consistent and also useful for scale-up processes. 27
Determination of Oxygen-Absorption Rate
The oxygen absorption rate per unit volume qa/v can be
estimated by
must
maximize

Since the oxygen is sparingly soluble gas, the overall mass-

transfer coefficient KL is equal to the individual mass-transfer
coefficient kL

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Methods of KLa determination

Sodium Sulfite Oxidation Method (1)

The sodium sulfite oxidation method (Cooper et al., 1944) is
based on the oxidation of sodium sulfite to sodium sulfate in
the presence of catalyst (Cu2+ or Co2+) as

To measure the oxygen-transfer rate in a fermenter, fill the

fermenter with a 1 N sodium sulfite solution containing at least
0.003 M Cu2+ ion. Turn on the air supply and start a timer when
the first bubbles of air emerge from the sparger. 30
Allow the oxidation to continue for 4 to 20 minutes, after which,
stop the air stream, agitator, and timer at the same instant, and take
a sample.

Mix each sample with an excess of freshly pipetted standard iodine

reagent. Titrate with standard sodium thiosulfate solution
(Na2S2O3) to a starch indicator end point.

Once the oxygen uptake is measured, the kLa may be calculated by

using following equation, where CL is zero and CL* is the oxygen
equilibrium concentration.

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Limitations of sulfite oxidation method

• The solution cannot approximate the physical and chemical

properties of a fermentation broth.
• An additional problem is that this technique requires high
ionic concentrations (1 to 2 mol/L), the presence of which
can affect the interfacial area and, in a lesser degree, the
mass-transfer coefficient

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Example 9.6
To measure kLa, a fermenter was filled with 10 L of 0.5 M sodium sulfite solution
containing 0.003 M Cu2+ ion and the air sparger was turned on. After exactly 10 minutes,
the air flow was stopped and a 10 mL sample was taken and titrated. The concentration
of the sodium sulfite in the sample was found to be 0.21 mol/L. The experiment was
carried out at 25°C and 1 atm. Calculate the oxygen uptake and kLa.

Solution: The amount of sodium sulfite reacted for 10 minutes is 0.5 - 0.21 = 0.29
mol/L.

Acc to stoich, the amount of oxygen required to react is 0.29 mol/L Na2SO3 x ½ mol O2
/ mol Na2SO3 = 0.145mol/L.
*Therefore, oxygen uptake qa/v is

From Henry’s law, solubility of oxygen in equilibrium with air can be estimated by:

P(gas 1) = x(gas 1) * P(Total)

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H. The value of Henry's law constant at T can be obtained from the solubilities listed
in Table 9.2

Po
C* =
H
For example, at 25°C, CL* is 1.26 mmol/L and pO2 is 1 atm because it
is pure oxygen. By substituting these values into Eq.
we obtain HO2 (25°C) is 0.793 atm L/mmol.
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Dynamic Gassing-out Technique (2)

This technique monitors the change of the oxygen

concentration while an oxygen-rich liquid is deoxygenated by
passing nitrogen through it. Polarographic electrode is usually
used to measure the concentration. The mass balance in a vessel
gives

from which kLa can be calculated based on the measured values of CL(t1)
and CL(t2)
Direct Measurement (3)
In this technique, we directly measure the oxygen content of
the gas stream entering and leaving the fermenter by using
gaseous oxygen analyzer. The oxygen uptake can be calculated
as

Once the oxygen uptake is measured, the kLa can be

calculated by using (below) equation where CL is the oxygen
concentration of the liquid in a fermenter and C*L is the
concentration of the oxygen which would be in equilibrium
with the gas stream.
The oxygen concentration of the liquid in a fermenter can be
measured by an on-line oxygen sensor.
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 Dynamic Technique

By using the dynamic technique (Taguchi and Humphrey,

1966), we can estimate the kLa value for the oxygen transfer
during an actual fermentation process with real culture
medium and microorganisms. This technique is based on the
oxygen material balance in an aerated batch fermenter while
microorganisms are actively growing as

While the dissolved oxygen level of the fermenter is

steady, if you suddenly turn off the air supply, the oxygen
concentration will be decreased with the following rate
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since kLa in Eq. is equal to zero. Therefore, by measuring the
slope of the CL vs. t curve, we can estimate rO2CX . If you turn
on the airflow again, the dissolved oxygen concentration will
be increased according to above equation which can be
rearranged to result in a linear relationship as
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Reference

Lee, J. M. (2009). Biochemical engineering.

Englewood Cliffs, NJ: Prentice Hall

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n)

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The different flow patterns produced by a disc
turbine that occur under a range of aeration and
agitation conditions (Fig. 9.13).

Fig 9.13A shows the flow profile of a non- aerated vessel and Fig9.13B to F
the profiles with increasing airflow rate. As air-flow rate increases the flow
profile changes from one dominated by agitation (Fig. 9.13 B)to one
dominated by air flow (Fig9.13 D to F) until finally the air flow rate is such that 45

the air escapes without being distributed by the agitator (F)

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Quantitative relationships between power
consumption and operating variables may be
useful in:
i. Estimating the amount of power that an
agitation system will consume under certain
circumstances, which could assist in fermenter
design.
ii. In providing similar degrees of power
consumption (and, hence, agitation and,
therefore, KLas in vessels of different size

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 1. MEDIUM RHEOLOGY

Fermentation media contain starch as a carbon source which

may render the medium non-Newtonian and relatively
viscous. As the organism grows it will degrade the starch and
thus modify the rheology of the medium and reduce its
viscosity.

The study of the growth of Streptomyces aureofaciens on a

starch-containing medium.
▪ Before inoculation, the medium displayed Bingham plastic
characteristics with a well-defined yield stress and an
apparent viscosity of approximately 18 pseudopoise;
▪ after 22 hours the organism's activity had decreased the
medium viscosity to a few pseudopoise and modified its
behaviour to that of a Newtonian liquid;
▪ from 22 hours onwards the apparent viscosity of the broth
gradually increased, due to the development of the
mycelium, up to a maximum of approximately 90
pseudopoise and the rheology of the broth became
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increasingly pseudoplastic in nature.
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The difference in the pattern of oxygen uptake between unicellular and mycelial
fermentations.
In both unicellular and mycelial fermentation the pattern of total oxygen uptake is very
similar during the exponential growth phase, up to the point of oxygen limitation.

However, during oxygen limitation, when arithmetic growth occurs, the oxygen uptake
rate remains constant in a unicellular system whereas it decreases in a mycelial one.
The only possible explanation for such a decrease is the increasing viscosity of the
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culture caused by the increasing mycelial concentration.
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▪ The upper graph represents the relationship
between the dissolved oxygen concentration and
the volumetric oxygen transfer rate achievable in
three fermenters (plots 1, 2 and 3 represent
fermenters of increasing KLa values) while

▪ the lower graph represents the relationship

between biomass and the volumetric oxygen
uptake rate of the culture.

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Reference

Nongkynrih, M and Sir, P (2014). Aeration and

Agitation

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