Industrial Microbiology

INDM 4005

Lecture 12

27/02/04
 Continuation of Lecture 11
5.1.4. Factors influencing oxygen supply
5.1.4. (a) process factors
5.1.4. (b) transfer through an interface (KLa)
5.1.4. (c) determination of KLa;
5.1.4. (d) factors affecting bubble size
5.1.4. (e) gas hold-up :
5.1.4. (f) economics of oxygen transfer
 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 O2 can

be delivered to the biological sysytem (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
 Oxygen Transfer Rate
• Rate of O2 transfer from air bubble to the liquid phase may be described as

dCL = KLa (Csat - CL)

dt
CL = Is the concentration of dissolved O2 in broth in mmoles L-1
t = Is time in hours
dCL = Is the change in O2 conc over time in mmoles O2 L-1 hr-1
dt
-1
KL = Is the mass transfer coefficient gas to liquid phase, (cm h )
a = Is the gas/liquid interface area per liquid volume (cm2 cm-3)
Csat = Is the saturated dissolved oxygen conc in mmoles dm-3
 KLa
KL = Is the mass transfer coefficient (cm h-1)
a = Is the gas/liquid interface area per liquid volume (cm2 cm-3)

These are difficult to measure individually and are generally

linked to give

KLa = volumetric mass transfer coefficient per hr

Therefore KLa is a measure of the aeration capacity of a

fermenter and must be maintained above a minimum
critical level to satisfy oxygen requirements
 5.1.4.(d) FACTORS AFFECTING BUBBLE SIZE
(a) Influence of gas velocity on bubble formation:

slow medium fast

.
:::
::
.
..
::..
...
..
low medium ::..
high
Very little backmixing
backmixing
slug flow
Slug Flow - flow alternates between high-liquid and high-gas composition.
• b) Influence of liquid properties on bubbles;

Two types of liquid

B bubbles coalesce

Therefore given same aeration equipment

B will give a greater range of bubble size

A B

Liquid can change from A  B when salts are added. Implication

for mass transfer in different media. Will this property of liquids
influence Kla - why?
 5.1.4.(e) GAS HOLD-UP
Represents air volume retained in the liquid

Vh = V - V0
Where Vh = hold-up volume, V = vol. of gassed liquid, V0 = vol
of ungassed liquid.
No air
Air

Difference in volume represents hold-up volume

That is the amount of gas retained in the liquid
Correlations exist that relate hold-up to power input , for
example,

(P/V)0.4 . Vb 0.5
P/V = power input per unit vol ungassed liquid, V = linear velocity of air bubble
(ascending velocity). b

Ascending velocity of bubble (Vb):

V = FH /H V
b l 0

Where H0 = hold-up of bubble, F = aeration rate, Hl = liquid

depth, V = liquid volume.
CASE STUDY:
Review the literature on determination of gas hold up.
Show an application of optimising hold-up in a reactor
i.e. through mixing or length to width ratio (increasing
path length)
How does height (h) of a reactor vary with radius (r)
when volume (v) is kept constant?

volume of a cylinder is v = r 2 h
Let us fix the volume as 1 then
h = 1/  r 2
If r = 1 then h = 1/
r = 2 then h = 1/4
r = 3 then h = 1/9

Therefore as the radius increases the height (or path

length) decreases as the square of the radius
 5.1.4.(f) ECONOMICS OF OXYGEN
TRANSFER
Fermentation e.g Penicillin - high KLa
Waste treatment - economy

Kla . Csat = maximum rate at which oxygen can be

dissolved

Economy and capacity related through power input

per unit volume (P/V)

ECONOMY = KLa. Csat / (P/V)

CASE STUDY
Compare a pumped air, sparged system of
aeration with a surface aerator (as used in
waste treatment i.e.

What are the advantages of each system?

The balance between OXYGEN DEMAND and SUPPLY

Must consider how processes may be designed such that O2 uptake rate of
the culture does not exceed the oxygen transfer rate of the fermenter

Uptake rate = QO2.X

QO2 = O2 uptake rate, X = Biomass
dC/dt = KLa(Csat - CL ) = supply rate
Dissolved O2 conc. should not fall below the critical dissolved O2 conc
(Ccrit)
A fermentation 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
Mixing and agitation
 Overview
5.2. Mixing and agitation
5.2.1. Measurement of mixing
5.2.2. Function of agitation - mixing assists oxygen transfer
5.2.3. Mixing patterns arising from agitator design /agitation etc
5.2.4. Types of impellers
5.2.5. Impeller design and reactor design
5.3. Fluid dynamics
5.3.1. Fluid shear rate
5.3.2. Non-newtonian fluids
5.3.3. Reynolds number (nre)
5.3.4. Reynolds number of an agitator
5.3.5. Relationship between power consumption and operating
variables
5.4. Agitation / aeration and scale-up
 5.2. MIXING and AGITATION
5.2.1. Measurement of mixing;

Measure degree of mixing using residence time

distribution (RTD) of a tracer dye e.g.

Residence time distributions are important when

calculating scale-up factors

(Is the mixing of equal quality in two different sizes

/types of fermenter?)
 Residence time distributions are important when calculating scale-up factors

Measure conc of dye

Up-flow reactor with substrate

added at the base of the reactor,
Add dye as product etc exits at the top
a pulse
Examine pattern of appearance of the pulse of dye in exit stream I.e
plot concentration versus time

T T T T
Plug flow Normal Channelling Back mixing
 5.2.2. FUNCTION OF AGITATION -
Mixing assists Oxygen transfer
(a) Increases interfacial area (between gas and liquid) by
dispersing air in the form of small bubbles
(b) Increases hold-up time of bubbles in the liquid
(c) Prevents coalescence of air bubbles
(d) Decreases thickness of interfacial area

The degree of agitation in a STR is a function of;

 design of agitator,
 Rheology / power /energy input
 5.2.3. MIXING PATTERNS ARISING FROM AGITATOR
DESIGN /AGITATION etc.

Axial flow

Radial flow
Propeller Impeller

Influence
of baffles

Vortex fermenter
High Density Low Density
Bubbles impart velocity

Air
 5.2.4. TYPES OF IMPELLERS

 Disc turbine - most usual,

 vaned disc
 open turbine (variable pitch)

CASE STUDY:
Draw each type of impeller
P125 Stanbury & Whitaker
5.2.5. IMPELLOR DESIGN AND REACTOR DESIGN
Impellor diameter and spacing
Rotational speed (tip speed)
Geometry e.g. size of blade
Position of impellor
 Spacing:
Di < Hi, 2Di
 Number
(HL - Di)/Di > Number of impellors > (HL - 2Di)/Di
Di = impellor diameter
Hi = spacing
HL= height of liquid in fermentor

Relationship between diameter and tip speed - see

REYNOLDS NUMBER
This influences the level of shear experienced by a cell.
 Case study
Design an impeller type agitation system for a 1000l
fermenter used to grow bacteria
 5.3. FLUID DYNAMICS

• In liquid culture the fluid flow or rheological behaviour

have a major impact on mass transfer
• Fluids may be described as Newtonian or non-Newtonian
depending on whether their rheology (flow) characteristics
obey Newtons law of viscous flow

Fluid viscosity = shear stress or force per unit area

shear rate or velocity gradient
 A rheogram of a Newtonian fluid
Plot of shear rate against shear stress

Shear
stress
Slope = viscosity

Shear rate
The viscosity of a Newtonian fermentation broth will not vary
with agitation rate
 Rheograms of fluids of different rheological properties

Bingham plastic

Newtonian fluid
Shear
Casson
stress body

Dilatant Pseudoplastic

Shear rate

However, a non-Newtonian liquid which does not obey

Newtons law of viscous flow will vary depending on shear rate
i.e agitation rate
5.3.1. FLUID SHEAR RATE:

EQUAL POWER

Large and slow Small and fast

PUMPING
VELOCITY HEAD
Ratio of velocity head to pumping + shear rate of fluid (is the
velocity gradient)

This concept important when dealing with large cells subject to

shear damage  plant cells require large, slow impeller i.e.

Bacteria Plant cell

Moves with the Deformed (ruptured)

liquid layers by velocity gradient
 Fluid shear rate
Fluid shear stress (= applied force) is Viscosity X Fluid shear
rate (ratio of shear stress to shear rate = viscosity in
Newtonian fluid). In a general way one can think of viscosity
as "resistance to mixing" (i.e stir a jar of molasses compared
with jar of water !)

See plots of Shear stress to shear rate for Pseudoplastic

(viscosity decreases as force is applied), Bingham plastic
(once fluid yields viscosity is constant) , Newtonian (viscosity
is constant - i.e. slope).
Case study:
Outline the problems encountered in
growing fragile cells and what strategies
are used to optimise productivity
 5.3.2. NON-NEWTONIAN FLUIDS
Many fermentation broths (e.g. mycelial, polymer production etc.) are
NON-NEWTONIAN FLUIDS (like Bingham plastic or Casson).
Viscosity significantly alters with time (e.g. t0 approaches Newtonian, t100
is Non-Newtonian). See the effect of P. chrysogenum mycelia on Kla.

100
Effect of
Penicillium crysogenum
Kla mycelia on Kla (Fig 9.14)
(% of
original)

0
1.5%(w/v)
Mycelium conc
 Culture alters physical aspects such as mass transfer
 5.3.3. REYNOLDS NUMBER (NRe)
Relates to flow of liquids (motion).
Flow of liquid over a stationary surface may be considered as
movement of an infinite number of fluid layers each moving with a
velocity that increases with distance from the surface

Streamline / laminar Turbulence

Reynolds number is a function of the ratio of turbulent to viscous (fluid) force.

i.e
NRe = (linear velocity x density x linear dimension) / viscosity
can be used when any force is applied to a liquid.
 5.3.4. Reynolds number of an agitator
NRe is a function of the ratio of turbulent to viscous
flow

NRe = (ND2 )/v

N= agitator speed, D = diameter of impellor,  = density

of liquid, v = viscosity.

Low number ( 1 - 15) = laminar flow

up to approx 3 x 10 3 = transitional range

above = turbulence
Example: Fermenter system type of agitation for (a)
water and (b) polymer

(a) Water;
viscosity = 0.001(1 centipoise)
 NRe = ND2 x 100
Low N or D values give NRe > 3 x 10 3 (turbulence -
good mixing)

(b) Polymer;
viscosity = 100 (10,000 centipoise)
 NRe = ND2  100
Much higher N or D required to give values > 3 x 10 3
(i.e. turbulence)
 5.3.5 Relationship between power
consumption and operating variables
Power number = Np = P / (N3D5)
Represents power absorbed during agitation of non-
gassed liquids.

Plot of Np versus NRe (log - log plot) gives the Power

curve which is divided into 3 regions
 Laminar or viscous flow
 Transition zone
 Turbulent zone - normally this is the required
state
From this, one can predict the necessary power to give a
specific level of mixing (or adequate mixing - should be
relatively constant at NRe> 4 x 10 3)

This Power curve developed by Rushton used ungassed

systems thus not typical of aerators - aeration contributes
to mixing.
Michel and Miller developed an empirical correlation for
gassed power consumption -

Pg = k (P2ND3/Q0.56) 0.45 this incorporates a function

representing aeration rate and can be used to predict power
consumption in gassed systems. More recent correlations
are also available e.g Humark (1980).
Correlations between KLa and power exist for example,
KLa = k(Pg/V) 0.95 Vs0.67
the figure 0.95 can vary with scale 0.95 - Lab, 0.67 - Pilot, 0.5
- Plant.

This incorporates parameters such as liquid volume (V),

superficial air velocity (Vs) and if one substitutes the Michel
and Miller equation for Pg one then relates to impellor design
(N and D) and the volumetric air flow rate (Q).

Also illustrates how Kla is affected by consumption of power

and degree of agitation.
 5.4. AGITATION / AERATION and
SCALE-UP

shear
cost
agitation
CO
2

foam
oxygen
mixing

aeration
This illustrates the "scale-up" window defining the operating
boundaries for aeration and agitation in the scale-up of a
typical fermentation.

Problems that may arise:

ACTION RESULT
Minimise aeration CO2 and O2 levels
Maximise aeration Foam formed
Minimise agitation Bulk mixing poor
Maximise agitation shear, cost increased
For each of the following statements, select appropriate methods of
KLa determination from the list provided.

1) The KLa value is considered a characteristic of the bioreactor

rather than the fermentation process Sulphite reduction

2) Requires measurement of gaseous O2 content Oxygen balance

3) Requires multiple measurement of dissolved O2 concentration

Dynamic gassing out
4) Only one parameter has to be measured Dynamic gassing out

5) O2 saturating concentration must be known Oxygen balance and sulphite

6) Method influenced by response time of dissolved oxygen sensor

Dynamic gassing out
Oxygen balance, sulphite reduction, dynamic gassing out
Page 193, In vitro cultivation of microorganisms
 Summary
• Agitation of suspended cell fermentations is performed to mix the
three phases within a fermenter

• Transfer of O2 into liquid broth is enhanced by agitation

• Agitation promotes retention of air bubbles in suspension, reduces

bubble size, prevents bubble coalescence

• Important to get correct shear conditions

• Fermenter agitation requires substantial input of power

• Understand Newtonian and non-Newtonian liquids

• Understand Reynolds and Power Numbers, Power curve