CHAPTER 4

MASS TRANSFER
BIOPROCESS TECHNOLOGY
BSB 3123

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§ To understand the theory of
diffusion.

§ To understand the theory of

oxygen transfer in bioprocessing.

§ To be able to calculate kLa value.

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 CONTENTS

4.1 Role of Diffusion in Bioprocessing

4.2 Film Theory
4.3 Oxygen Uptake in Cell Cultures
4.4 Oxygen Transfer From Gas Bubble to Cell
4.5 Oxygen Transfer in Fermenter
4.6 Measurement of kLa

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When dye is dropped into a pail of water, mass
transfer processes are responsible for movement of dye
molecules through the water until equilibrium is
established and the concentration is uniform

Mass transfer dependent on concentration gradient in

the system

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§ Yellow food coloring diffusing through water. The glass on the left contains hot
water, while the glass on the right contains cold water. The food coloring was added
to the cold water slightly before the coloring was added to the hot water, yet after a
few seconds it has diffused more thoroughly through the hot water. Why?

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 Molecular Diffusion
Molecular diffusion – movement of component molecules
in a mixture under the influence of a concentration difference
in the system

Diffusion of molecules occurs in the direction that required

to destroy the conc. gradient

Role of Diffusion in Bioprocessing

Fluid mixing is carried out in most industrial processes where
mass transfer takes place

Areas of bioprocessing in which diffusion plays a major role

are described in the following:
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 Mixing on a molecular scale
Scale of therefore relies on diffusion as
mixing the final step in the mixing
process

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 - In biological system: solid form
(clumps, flocs, film)
Solid phase - Solid particle: substrate must be
reaction transported into solid

- The mass transfer mechanism:

molecular diffusion

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 -When different phases come into contact
: fluid velocity near the phase interphase
decreased

- Mass transfer: diffusion across the phase

Mass transfer interphase
across a phase
boundary - Example:
1) O2 transfer from gas bubble to
fermentation broth

2) Penicilin recovery from aqueous to

organic liquid

3) Glucose transfer from liquid medium to

mould pellets

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 Film Theory

Mass transfer of solute from one phase to another involves

transport from the bulk of one phase to the phase
boundary/interface

Then from the interface to the bulk of the second phase.

The film theory – based on the idea that a fluid film or mass
transfer boundary layer forms wherever there is contact
between 2 phases

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Figure 4.1 Film resistance to mass transfer between 2 immiscible liquids

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 Oxygen Uptake in Cell Cultures
Cell in aerobic culture take up oxygen from the liquid

The rate of oxygen transfer from gas to liquid is importance, especially at

high cell densities when cell growth is likely to be limited by availability of
oxygen in the medium

An expression for rate of oxygen transfer from gas to liquid is given by

the following equation (1);

N A = kLa C ( *
AL - C AL ) Eq. (1)
NA :rate of oxygen transfer per unit volume of fluid (gmol m-3s-1)
kL : liquid-phase mass transfer coefficient (ms-1)
a : gas-liquid interfacial area per unit volume of fluid (m2 m-3)
C*AL : oxygen concentration in broth in equilibrium with the gas phase (gmol m-3)
CAL : oxygen concentration in the broth (gmol m-3)

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 QUESTION
1. A fermentation system has a kLa of 3/s and a C* of 5 ppm of O2.
if the bulk liquid is saturated with oxygen. What is the OTR?

2. A fermentation system has a kLa of 3/s and a C* of 5 ppm of O2.

If the bulk liquid is completely depleted with oxygen. What is
the OTR?
 Factors Affecting Oxygen Demand
The rate at which oxygen is consumed by cells in fermenters determines
the rate at which it must transferred from gas to liquid

Many factors affecting oxygen demands such as:

a) Cell species
b) Culture growth phase
c) Nature of the carbon source in the medium

In batch culture oxygen uptake varies with time due to:

a) Concentration of cells increases during the course of batch culture
b) Total rate of oxygen uptake is proportional to the number of cells present

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 Oxygen Transfer From Gas Bubble to Cell

In aerobic fermentation, oxygen molecules must overcome a series of

transport resistances before being utilized by the cells

8 mass-transfer steps involved in transport of oxygen from the interior of

gas bubbles to the site of intracellular reaction :

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 8 mass-transfer steps involved in transport of oxygen from the interior of
gas bubbles to the site of intracellular reaction :

1) Transfer from the interior of the bubble to the gas-liquid interface

2) Movement across the gas-liquid interface
3) Diffusion through the relatively stagnant liquid film surrounding the
bubbles
4) Transport through the bulk liquid
5) Diffusion through the relatively stagnant liquid film surrounding the cells
6) Movement across the liquid-cell interface
7) If the cells are in a flocs, clump or solid particle, diffusion through the
solid to the individual cell
8) Transport through the cytoplasm to the site of reaction

* If the cells are individually suspended in liquid rather than in a clump, step
(7) is omitted
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Figure 4.1 Steps for transfer of oxygen from gas bubble to cell
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Analysis for most bioreactors in each step involved

1) Transfer through the bulk gas phase in the bubble is relatively fast
2) The gas-liquid interface itself contributes negligible resistance
3) The liquid film around the bubbles is a major resistance to oxygen transfer
4) In a well mixed fermenter, concentration gradients in the bulk liquid are
minimized and mass transfer resistance in this region is small, except for viscous
liquid
5) The size of single cell <<< gas bubble, thus the liquid film around cell is thinner
than that around the bubble. The mass transfer resistance is negligible, except
the cells form large clumps
6) Resistance at cell-liquid interface is generally neglected
7) The mass transfer resistance is small, except the cells form large clumps or flocs
8) Intracellular oxygen transfer resistance is negligible because of the
small distance involved
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 The relatives magnitudes of the various mass-transfer resistance depend
on the composition & rheological properties of the liquid, mixing
intensity, bubble size, cell clump size, interfacial adsorption
characteristics and other factors.

When cells are dispersed in the liquid and the bulk fermentation broth is
well mixed – the major resistance to oxygen transfer is the liquid film
surrounding the gas bubbles.

Consequently, the rate of oxygen transfer from the bubble all the way to
the cell is dominated by the rate of step (3)

The mass transfer rate for this step can be calculated using Eq. (1)
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 At steady state there can be no accumulation of oxygen at any
location in the fermenter

So, the rate of oxygen transfer from bubbles must equal to the rate of
oxygen consumption by the cells

N A = qO x
From Eq. (1), we obtain the following equation (2):

(
k L a C - C AL = qO x
*
AL ) Eq. 2

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 kLa – used to characterize the oxygen mass transfer capability in
fermenters.

If kLa is small – the ability of the reactor to deliver oxygen to the cells is
limited

For a given set of operating conditions, the maximum rate of oxygen

transfer occurs when the concentration-difference driving force (C*AL - CAL)
is highest; when CAL is zero.

Therefore from Eq. (2), the maximum cell concentration can be:

*
k L aC
xmax = AL
qO Eq. 3

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 Example 4.1 Cell concentration in aerobic culture

A strain of Azetobacter vinelandii is cultured in a 15 m3 stirred fermenter for

alginate production. Under current operating conditions kLa is 0.17 s-1 .
Oxygen solubility in the broth is approximately 8 x 10-3 kg m-3 .

a) The specific rate of oxygen uptake is 12.5 mmol g-1 h-1 . What is the maximum
possible cell concentration?

b) The bacteria suffer growth inhibition after copper sulphate is accidently added to
the fermentation broth. This causes a reduction in oxygen uptake rate to 3 mmol g-1
h-1 . What is maximum cell concentration can now be supported by the fermenter?

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 Solution:
From Eq. (3): *
k L aC AL
xmax =
qO
(a) xmax =
(0.17 s )(8 ´10
-1
kgm -3 -3
)
12.5mmol 1h 1gmol 32 g 1kg
× × × ×
gh 3600 s 1000mmol 1gmol 1000 g

= 1.2 ´10 4 gm -3 = 12 gl -1

(b) Assume that addition of copper sulphate does not affect C*AL or kLa. If qO is
reduced by a factor of 12.5/3 = 4.167; xmax is increased to:

xmax = 4.167 (12 g l-1) = 50 g l-1

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 Oxygen Transfer in Fermenters

The rate of oxygen transfer in fermentation broths is influence by several

physical and chemical factors that change either the value of kL or the value of
a, or the driving force for mass transfer (C*AL - CAL)

The factors are:

1. Bubbles
2. Sparging, stirring, and medium properties
3. Antifoam agents
4. Temperature
5. Gas pressure and oxygen partial pressure
6. Presence of cells

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 Measurement of kLa

kLa can be determined experimentally

– dynamic method

This method based on an unsteady-state mass balance

for O2

The main advantage of the dynamic method over the

steady-state technique is the low cost of the equipment
needed

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Dynamic method:

1. The fermenter contains cell in batch culture

2. At some time t0 the broth is de-oxygenated either by sparging N2
into vessel or by stopping the air flow if the culture is oxygen-
consuming
3. Dissolved oxygen (DO) concentration CAL drops during this period
4. Air is then pumped into the broth at a constant flow rate and the
increase in CAL monitored as a function of time
5. Ensure the oxygen concentration remains above Ccrit so that the
rate of oxygen uptake by the cells is independent of oxygen level

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Figure 4.2 Variation of oxygen tension for dynamic measurement of kLa
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 During the re-oxygenation step, the system is not at steady state

The rate of change of DO:

= k L a (C AL - C AL ) - qO x
dC AL *

dt Eq. (4)
qox : rate of oxygen consumption

When,
dC AL ,
C AL = C AL , dt = 0

because there is no change in CAL with time, therefore:

qO x = k L a C ( *
AL - C AL ) Eq. (5)

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 Substitute Eq. (5) into Eq. (4) gives:

dC AL
dt
(
= k L a C AL - C AL ) Eq. (6)

Assuming kLa is constant with time, then integrate Eq.(6) gives:

æ C AL - C AL1 ö
lnçç ÷
÷ Eq.(7)
è C AL - C AL 2 ø
kLa =
t 2 - t1

æ C AL - C ö
kLa can be estimated when plotting lnç AL1 ÷
versus (t 2 - t1 )
ç C AL - C AL ÷
è 2 ø

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Figure 4.3 Evaluating kLa using the dynamic method
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Example 4.2 Estimating kLa using dynamic method

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Thank
you

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