ECH4202

Bioreactor Engineering Design

Topic 8: Scale-up
29 May 2023
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 Learning objectives
• To explain the basis of scale up and its effect in
heat and mass transfer in bioreactor
• To apply geometry similarity and non-
similarity method for bioreactor scale up

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 Scale up success story
• Scale-up is the key for the commercial success

Penicillin
• Process developed in England
• Scaled up in the US (up to 400 m3, stirred power 5 kW/m3)
• Commercial success by the US
High Fructose Corn Syrup
• Process developed in the US
• Scaled up in Denmark
• Commercial success by Denmark
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Example: Penicillin Production

• Strain: Penicillium
chrysogenum
• Substrate: corn steep
liquor, lactose, etc
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Breakthrough in Penicillin production
• Improvement of culture media
• Isolation of better strains Penicillium chrysogenum
• Development of submerged culture techniques
• Development of mutant strains (Old: X-ray, UV-
irradiation, new: cloning, recombineering, CRISPR).
• Addition of precursors for penicillin synthesis
• Better separation techniques

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 Three Myths of Scale-Up
• We can just make it bigger
• Technology is already there
• It can be done very quickly

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 Why Scale-up is a challenge?
• Difficult to maintain homogeneity on large scale
– Microbes experience gradients at large scale:
temperature, O2, dissolved concentration, pH,
substrate
• Culture has more pressure to mutate with number of
divisions (phenotype changes)
• Phage contamination
• Use of industrial-grade nutrients
• Changes in medium due to length of sterilization
time 8
 Factors impacting Scale-up
Physical factors Process factors
• Mass & heat transfers • Pre-culture conditions
• Mixing • Sterilization quality
• Power • Inoculation ratio
• Shear stress • Bioreactor design
• pH, temperature, DO • Separation
• Cost estimations
Biochemical factors
• Media components
• Concentration of nutrients
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Genetic instability

Increasing number of
generations as size of the
fermenter increases

No x 2n = 32No cells
n = 5 (generations)

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Genetic instability increases as n increases

after few
Zainuddin HS, 2022
generations
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Zainuddin HS, 2022
Solution requires elaborate studies
Solutions
• Plasmid / genetic types
• Temperatures, pH
• Addition of selective
pressure (antibiotics)
• Nutrient complementation
• Inoculation ratio
• Media supplement

Bryskin A. V. et al, Plos One, 2010

Wein T et al, Nature Comm., 201912

 Approach for Scale-up
• Find geometric parameters of the • Cylindrical Tank
larger scale bioreactor
• Agitation and aeration
– Stirred tank reactor Vessel
• Cell suspension
diameter, height,
• Impeller diameter • Enhance the aeration
– Bubble column Height, • (oxygen limitation problem)
Diameter • Mixing
• Use scale up criteria to determine • Problem with shear-
operational parameters of the larger sensitive cells
scale • Heating and cooling
– Stirred tank reactor • Sensors
• Impeller speed, Air flow rate • pH Control
– Bubble column Air
flow rate 13
Geometry of STR
For Turbine blade impellers

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 Scale-up criterion: Preservation of
geometrical similarity
• Transfer experimental data from lab and pilot plant equipment
to large scale industrial equipment

• Ideal scale criterion: parameter that has same numerical value

as the volume of geometrically similar bioreactor increase in
size
HL1 = H L 2 = H L 3
Dt1 Dt 2 Dt3
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 Dynamic scale-up criteria
1. Constant volumetric mass transfer coefficient
(KLa)
2. Constant power consumption per unit
volume (P/V)
3. Constant impeller tip speed (πND)
4. Constant Reynolds number (Re)
5. Constant dissolved oxygen concentration
(CO2)
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 Dynamic Scale-up Criteria

P/V (30%)
Kla(30%)

!ND(20%)

O2 con. (20%)

Percentage of each criteria used in fermentation

industry
(Garcia & Gomez, 2009)
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 Scale-up ratio from 2 L to 20 L

(Garcia and Gomez,2009)

T = tank height, P = Power, N=rpm, V=volume 18

(Aydin Berenjian, 2019) 19
 Constant volumetric mass transfer
coefficient (KLa)
• If KLa is selected as a scale-up criteria, then:
KLasmall = KLalarge

• Dependent variable used are:

1. Agitation power (P)
2. Aeration rate (Fg)
3. P/V
4. Tip speed (N)
5. Reynold number 20
Constant power consumption per unit
volume (P/V)
• Turbulent: P = k.ρ.N3.D5
• Laminar: P = k.ρ.N3.D2

• For laminar scale-up; P/V = constant

N3D2small = N3D2 large

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 Constant tip speed (!ND)
The tip speed is simply the distance that any selected point on
the peripheral of the impeller travels in a set time. In other
words the “speed” of that point. Unit is in distance/time (m/s).

N1=Impeller speed of small bioreactor

N2=Impeller speed of Largebioreactor
Di1= Impeller diameter of small bioreactor
Di2= Impeller diameter of large bioreactor

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 Constant Reynolds number

N = Impeller speed
D = Impeller diameter
ρL= Density of the liquid
μL=Viscosity of the liquid

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 Tutorial: Geometry similarity
After batch fermentation, system is dismantled and
some of the cell mass is suspended in the liquid
phase (2L), while some is attached to the reactor
wall in a thick film (0.3 cm). Analysis using a
radioactive tracer shows that 50% of the target
product (intracellular) is associated with each cell
fraction. The productivity of this reactor is 2g
product/L for 2 Litre reactor size. Find the
productivity for 20,000L reactor, given that both
reactors have a height-to- diameter ratio of 2 to 1.
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 Tutorial: Dynamic similarity
Consider scale-up of reactor from 10 L to 10,000 L.
Small reactor has H:D of 3:1. Di = 1/3D. Agitator
speed is 500 rpm and 3 Rushton impellers were
used. Assume mixing was laminar. Determine the
dimensions of the large fermenter and agitator
speed for:
a) Constant P/V
b) Constant impeller tip speed
c) Constant Reynolds number
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Tutorial: Scale up using production capacity
Lysozyme is an anti-bacterial and anti-fungal widely
present in animals, plans and microorganism. The
bioprocess parameters of recombinant strain of
Kluyveromyces lactis were studied in a lab-scale
fermenter giving 110 IU/mL of lysozyme within 43
h. Engineers plan to produce annually 3 x 1012 IU of
this enzyme. Thus, what should the dimension of
the bioreactor looks like if H/D = 3, 20% headspace
are assumed, WBF/DT = 1/10, and Di/DT = 1/5.
Fermentation are to be operated for 11 months per
year.
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End
A continuous bioreactor in figure 1 is to be Example 3
assessed in the scale up study. The small
bioreactor is 10 L and equipped with 3
impellers mounted in the vessel, which
create three compartments of perfectly
mixed liquid (C1, C2 and C3). The overall
mass transfer coefficient between each
compartment (H) is 0.43 s-1 for 10 L and
0.075 s-1 for 10,000 L. The desired glucose
concentration is set at 25mg/L. This set
point is measured the second
compartment. The growth of microbial
culture in the vessel obeys Monod growth
with first order rate constant, k1 = 0.05s-1.
Find concentration at first and third
compartment in 10 L and 10,000 L reactor. 26
 Cont.
• Mixing characteristic in each compartment:

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