TEMA 8.

AGITACIÓ

AGITATION CONCEPTS

secretion of
high substrate growth
highly viscous
concentration morphology
products

broth
rheology
low viscosity high viscosity

variable costs

energy input cooling costs

for agitation

viscosity
fluid mechanics/power requeriments
heat transfer/mass transfer

raw material preparation fermenter conditions product separation/recovery

pumping, heating, microorganisms dispersion pumping, heating, cooling,

cooling, mixing oxygen transfer solid/liquid separations

growth product formation morphology

viscosity

design, economics

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8.1. Reologia dels brous de cultiu

flow behaviour of fluids μa = τ · γ-1 apparent viscosity

τ = τ0 + K· γ

τ = K· γn n>1
τ = K· γn n<1

τ = μ · γ

γ

♦ Newtonians
ocultius de llevats i bacteris, unicel·lulars no massa concentrats
oStreptomyces noursei productor nistatina
♦ Pseudoplastics
ocultiu de Saccharomyces a concentracions >10% en sòlids
ocultius filamentosos de Streptomyces niveus, Penicillium i Aspergillus
oproductors de polisacàrids (Xanthomonas i Aureobasidium pullulans)
♦ Plàstics de Bingham
omedi amb midó - producció de kanamicina
omedi amb midó - producció estreptomicina per Streptomyces
ohidroxilació d'esteroids

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8.1. Reologia dels brous de cultiu

Changes in the flow behavior index and the consistency index of the
culture broth of C. acremonium M25 (Cephalosporin C fermentation)

http://www.rheology.or.kr/down/17-1(3).pdf

Shear sensitivity
+ animal cells
plant cells
algae
- yeast/fungi/bacteria

shear stress nature

• turbulent
• laminar
• associated with interfaces (during bubble formation, rise and rupture)

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8.2. Factors i efectes de l’esforç tallant

factors

1. Type of cell and species

2. Composition and thickness of cell wall when present
3. Size and morphology of cell
4. The intensity and nature of shear stress
5. Growth history, both short-term (e.g., starvation) and longterm
adaptation
6. Growth environment (pH, temperature, agitation intensity,
light in photosynthetic cultures)
7. Growth medium (trace elements, vitamins, carbon and
nitrogen sources)
8. Growth rate
9. Growth stage
10. Type and concentration of shear-protective agents if present

shear stress/shear rate

γ = kU g
a
bubble column reactor

k = 1, a = 1,000- 5,000 m-1

air-lift reactor
kU gr
γ=
A
1+ d
Ar
γ = 3,26 - 3,51 · 102 ·UgR + 1,48·104 ·UgR2
for: 0.004 < Ugr (m/s) < 0.06

Ad cross-sectional area of the downcomer

Ar cross-sectional area of the riser
Ugr = riser superficial gas velocity, m/s

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8.2. Factors i efectes de l’esforç tallant

stirred tank bioreactor

six-bladed Rushton turbine agitating a Newtonian fluid

0,3
γ average = 4,2N⎛⎜ D I ⎞ ·⎛ D I
⎟ ⎜
⎞
⎟
⎝ D T ⎠ ⎝ WI ⎠
0.5
⎛ρ⎞
γ = 3.3N D I ⎜⎜ ⎟⎟
1.5

⎝μ⎠

reduced
• specific protein production rate
(55%)
• final yield (67%)
distinct lag phase

Effect of Rushton turbine tip speed on protein

production by the mold Neurospora sitophila

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8.2. Factors i efectes de l’esforç tallant

Penicillium chrysogenum

impeller tip speeds of 2.5 → 6.3 m/s

↓ specific penicillin production rate

↓ mean length of the main hyphae
↑ mycelial fragmentation
↑ frequency of branching

Saccharomyces cerevisiae

impeller tip speed

1.43 m/s, 90% of the cells were dead in 25 h
0.66 m/s about 15% died over the same period

(a) Effect of Rushton-turbine

agitation speed on the specific
growth rate of Spodoptera
frugiperda cells in serum-free batch
culture. The aeration rate was
constant at 0.03 vvm. The variation
of shear stress with impeller speed
is also shown (dashed line)

(b) The effect of aeration rate on the

specific growth rate of S. frugiperda
Sf9 cells in serum-free batch
culture. The Rushton-turbine
agitation rate (two turbines) was
constant at 200 rpm (τ = 0.66 N m-2).

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8.2. Factors i efectes de l’esforç tallant

The effect of bubble diameter on the specific death rate of Spodoptera

frugiperda insect cells in bubble columns aerated at a constant volumetric
gas flow rate of 10 ml/min

The maximum pressure produced during bursting of

air bubbles of various sizes on the surface of water.
cell attachment

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8.2. Factors i efectes de l’esforç tallant

2% serum
effect of foetal-calf-serum
concentration and air-flow
rate on the death rate
constant of a mouse–
mouse hybridoma in air-lift
bioreactor

3% serum

Media additives associated Derivatized celluloses (e.g. methyl cellulose)

with cell protection
Derivatized starches (e.g. hydroxyethyl starch)
Dextrans
Pluronics
Poly(vinyl alcohol)s
Poly(vinyl pyrrolidone)
Poly(ethylene glycol)s
Proteins (e.g. serum albumin)
Serum

To decrease shear stress

(1) the aeration rate should be kept as low as possible

(2) the mean bubble size should larger than 7 mm, preferably about
10–20 mm;
(3) the location of the sparger should be such that the rising bubbles
do not interact with any impellers
(4) with airlift and bubble column reactors the aspect ratio should be
about 6 or 14
(5) when an impeller is used, maximum: 600 rpm ~ 1.6 m/s ~
1.0·kWm-3

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8.3. Disseny dels sistemes d’agitació

AGITATION DESIGN

Braun 300 L

standard fermenter geometry (stirred tank)

Wb/DT = 0.10 Wb

H/DT = 1.0 (1.0-1.25)

DI/DT = 1/3 (0.25-0.50)
C/DT = 1/3 (0.17-0.50)
∆C/DT = 1.0 (0.35-1.0)
LI/DI = 0.25 ; WI/DI = 0.2 H
∆C
LI

WI

DI C

DT

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8.3. Disseny dels sistemes d’agitació

Mixing
• Maintain uniform conditions in the v essel (solid, liquid,
gas concentration, Temperature, pH).
• Disperse bubbles throughout the liquid
• promote bubble break-up
• increase gas-liquid interfacial transfer
• Promote mass transfer of essential nutrients
• Prevention of sedimentation
• Securing of heat transfer
• Solubility of the nutrient's components which are less
soluble

Minimal limit of the mixer rotation

Sedimentation
"Dead" zones
"Flooding" effect

impellers
radial flow
perpindicular to the shaft
axial flow
parallel to the shaft

(a) Marine-type propellers

(b) Flat-blade turbine, Wi = Di/5
(c) Disk flat-blade turbine, Wi = Di/5, Di = Dt/3, Li = Di/4 (Rushton)
(d) Curved-blade turbine, Wi = Di/3
(e) Pitched-blade turbine, Wi = Di/8
(f) Shrouded turbine, Wi = Di/8 (Vaned disk)

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8.3. Disseny dels sistemes d’agitació

Disk flat-blade turbine (Rushton)

μ (apparent viscosity) < 50 cP, high N (rotational speed)
radial flow impeller
disc with 6-8 blades
the most common type

Pitched/inclined blade propeller ⇒ axial component

• superior performance compared to Rushton impellers (lower

energy outputs required for equivalent oxygen transfer)
• produce less shear stress

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8.3. Disseny dels sistemes d’agitació

Marine propellers ⇒ three blades, wide range of N,

high shearing effect at high rotational speeds

Ekato Intermig for mycelial cultures

· the mixer consists of two mixers
· axial + radial flows are generated
· decrease of the maximal shear forces
· requires less energy and lower gas through-put
to produce same mass transfer coefficient as
turbine
.

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8.3. Disseny dels sistemes d’agitació

anchors.helical ribbons
High μ
Anchors, helical ribbons :D/T >0.9
Lower speeds, vessels generally not baffled

Baffles

Re >300

vertical radially located plates

width is about 10% from the bioreactor vessel diameter
3-4 plates in the reactor
necessary to prevent the formation of a funnel
consumed electric power of the mixer increases by about 20%

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8.3. Disseny dels sistemes d’agitació

Mixing Effectiveness
Mixing time tm
time required to achieve specified degree of
homogeneity, starting from the completely tm = f(N-1)
segregated state

∆C
LI

WI

DI

dimensionless numbers
in agitated/aerated systems to characterise the design and
performance of the process, in a scale independent manner

P = agitator power (W)

DI = impeller diameter (m)
ρ = fluid density (kg m-3)
N = impeller speed (s-1)
μ = fluid viscosity (N s m-2)
Q = fluid flow rate (m3 s-1)
Qg = gas flow rate (m3 s-1)

ungassed, single phase fluids

Power Number
P
N P = 5
ρN3D I
to calculate the Mechanical Power (P) being transmitted to
the fluid by a turbine/impeller of a given design

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8.3. Disseny dels sistemes d’agitació

Reynolds number
2
ρND I
N Re =
μ
similar to applications in pipes, etc., indicates the degree
of turbulence experienced in a stirred tank reactor

Relationship of Power number and Reynolds number

Depends on the geometry

and agitator type
Geankoplis, 3rd ed.

Rushton Np = 5
Flat 6-blade

propeller

Reynolds Number ranges for Rushton turbine (1)

Re < 101 laminar flow
101 < Re < 104 transitional flow
Re > 104 turbulent flow

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8.3. Disseny dels sistemes d’agitació

Relationship has 3 phases –laminar, transition and turbulent

Np = K . (NRe)x
K = constant dependent de la geometria

NRe < 10 x = -1 P = K . μ . N2 . DI3

NRe > 10.000 x=0 P = K . ρ . N3 . DI5

Bioreactors - turbulent flow.

• power number is constant for a given impeller design
• if we know the impeller diameter and the rotational speed of the
impeller (both easy to measure) we can subsequently estimate the
mechanical power input to the bioreactor

P
N P = 5
ρN3D I

Relationship NP vs NI

non newtonian fluids

NRe' = ρ . N . DI2 / μa
μa = viscositat aparent

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8.3. Disseny dels sistemes d’agitació

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8.4. Estimació del coeficient kla en sistemes amb aeració

gassed fluids
gas hold-up (Hr, ε)
volume occupied by gas phase/total volume
ε = Hr = Vg / (Vg + Vl)

AIR BUBBLES,
gas phase, Vg

VT liquid phase, Vl

AIR

ρg = (1 - Hr) . ρ Np = Pg / N3·DI5·ρg

Pg / P = ρg / ρ = 1 - Hr Pg / P ~ 0.3 - 1

Superficial gas velocity = Ug , ms-1

Qg air flow rate at S.T.P.

Ug = 2
πDTI cross sectional area of reactor vessel
4

Aeration Number (Na) – Useful measure of the gas

dispersion capabilities of the impeller.

Qg
2
DI Qg
Na = = 3
N·D I N·D I
used to quantify the power ratio Pg/P as a function of aeration rate Qg

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8.4. Estimació del coeficient kla en sistemes amb aeració

Pg/P

4
Na ·102

Power input of several types of stirrer in Newtonian fluids, dependent on the

gas throughput. i= number of blades in disk Turbine stirrer.

Correlació Np - Na

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8.4. Estimació del coeficient kla en sistemes amb aeració

0.43
Michel i Miller ⎡ P2 N D 3 ⎤
Pg = m ⎢ 0.56
I
⎥
⎢⎣ Q g ⎥⎦

Typical Specific Power Consumptions (P/V) kW/m3

Mild agitation 0.1

Suspending light solids
Blending of low viscosity liquids
Moderate Agitation 0.4
Gas dispersion, liquid-liquid contacting
Some heat transfer
Intense Agitation 1.0
Suspending heavy solids, emulsification
Blending pastes, dough 4.0
Industrial-scale fermenters 0.5-5
Lab-Scale fermenters 5-10

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8.4. Estimació del coeficient kla en sistemes amb aeració

AERATION AND AGITATION

Bioreactor Types
1. Stirred tank with internal mechanical agitation (impellers, turbines)
2. Bubble columns - gas sparging for agitation
3. Loop reactors - mixing and liquid circulation are achieved by
• injected gas
• mechanical pump
• combination of the two

Empirical correlations for kla

stirred tank bioreactor
Correlations are based on:
• P/V (power i nput pe r unit volume)
typically represents th e d egree o f
agitation.
• Ug (superficial gas velocity) is
representative of the degree of
aeration vs the size of the vessel

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8.4. Estimació del coeficient kla en sistemes amb aeració

α
⎛ Pg ⎞ β
k L a =c⎜
⎜ V ⎟ ⎟ Ug
⎝ L ⎠
kLa = Vol. mass transfer coefficient, s-1
Pg = gassed power, W
VL = Liquid working volume of bioreactor vessel, m3
Ug = superficial gas velocity, ms-1
alpha and beta terms are constants defined by the
α= 0 - 1 particular reactor system.
ß = 0.20 - 0.70

Water at pH = 7, T = 25 ºC α= 0.4, ß = 0.5, c = 0.025 Rushton

c, α, β depend greatly on
• ionic strength (medium composition)
• T
• pH

Van’t Riet (Review 1979)

non-coalescing systems α= 0.70, ß = 0.20 , c = 0.002

coalescing systems α= 0.40, ß = 0.50 , c = 0.026
Smith, Van’t Riet and Middelton (wide variety of agitator ty pes, sizes
and DI/DT ratios )
non-coalescing systems α= 0.475, ß = 0.4, c = 0.02

coalescing systems α= 0.475, ß = 0.4, c = 0.01

Calderbank and Moo-Young for six flat-bladed disc turbines (Rushton)

coalescing systems α= 0.4, ß = 0.5, c = 0.025

non-coalescing systems α= 0.7, ß = 0.3, c = 0.0018

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8.4. Estimació del coeficient kla en sistemes amb aeració

bubble column reactor

sterile air used for agitation and

oxygenation

the agitation term becomes negligible,

kla = 0.0023 (Ug / dB )1.58

Ug = superficial gas velocity, ms-1
dB = bubble diameter, m

bubble rise velocity ~ 0.2 m/s

bubble diameter ~ 0.006 m

air-lift reactor

air introduced to create circulation internal to the reactor

kla =α . (Ug)ß

ß = 0.9 - 1.0

bubble rise velocity ~ 0.2 m/s

bubble diameter ~ 0.006 m

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