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BIOREACTOR

• A device in which living cells or enzyme


systems are used to promote biochemical
transformation of matter
• Uses:
– pharmaceutical industries
– Chemical industry
– Waste treatment
– Biomedical applications
Types
• Microbial fermenter- cell growth is used to
produce metabolites, biomass, transformed
substrates, or purified solvents
• Enzyme reactors (cell free)- immobilized
enzymes often used for fluid-fluid and
solid-fluid contractors
GENERALIZED VIEW OF
BIOPROCESS
RAW MATERIALS

UPSTREAM PROCESSES

Media Formulation
Inoculum Equipment
and
Preparation Sterilization
Sterilization

BIOREACTOR - FERMENTER

Reaction Kinetics Transport Phenomena Instrumentation


and Bioactivity and Fluid Properties and Control

DOWNSTREAM PROCESSES

Recovery and Waste Recovery,


Separation
Purification Reuse and Treatment

THE BOTTOM LINE

REGULATION ECONOMICS HEALTH AND SAFETY


TYPICAL BIOPROCESS
FLOW SHEET
PREPARATION
OF BIOMASS FOAM CONTROL pH CONTROL
Innoculum Stages Antifoam Addition Acid-Alkali Addition

PRODUCT RECOVERY

CELL SEPARATION
BIOREACTOR Intracellular
product
Extracellular
1). CELL DISTRUPTION product
2). PRODUCT EXTRACTION

Free Cells,
Immoblized Cells PRODUCT
or CONCENTRATION
PROCESS
Enzyme Bioreactor

PRODUCT
SEPARATION

PURIFICATION

STERILIZATION
DRYING

RAW MATERIAS Air FINAL PRODUCT


Nutrients and Reactants
in Aqueous Solution
(may contain insoluble
organic and/or inorganic
materials)
Basic Bioreactor Design
Criteria
 Microbiological and Biochemical Characteristics of the Cell System
(Microbial, Mammalian, Plant)
 Hydrodynamic Characteristics of the bioreactor
 Mass and Heat Transfer Characteristics of the Bioreactor
 Kinetics of the Cell Growth and Product Formation
 Genetic Stability Characteristics of the Cell System
 Aseptic Equipment Design
 Control of Bioreactor Environment (both macro-and micro-
environment)
 Implications of Bioreactor Design on Downstream Products
Separation
 Capital and Operating Costs of the Bioreactor
 Potential for Bioreactor Scale-up
Summary of Bioreactor Systems
__________________________________________________________
Bioreactor Cell Systems Used Products

__________________________________________________________
 Air-Lift Bioreactor Bacteria, Yeast and SCP, Enzymes, Secondary
other fungi metabolites, Surfactants

 Fluidized-Bed Immobilized bacteria, Ethanol, Secondary


Bioreactor yeast and other fungi, metabolites, Wastewater
Activated sludge treatment

 Microcarrier Immobilized (anchored) Interferons, Growth factors,


Bioreactor mammalian cells on Blood factors, Monoclonal
solid particles antibodies, Vaccines, Proteases,
Hormones

 Surface Tissue mammalian, tissue Interferons, Growth factors,


Propagator growth on solid surface, Blood factors,
tissue engineering Monoclonal antibodies,
Vaccines, Proteases, Hormones

__________________________________________________________
Summary of Bioreactor Systems (Cont’d)
____________________________________________________________________________________________________
Bioreactor Cell Systems used Products
Design
________________________________________________________________________________________

 Membrane Bioreactors, Bacteria, Yeasts, Ethanol, Monoclonal anti-


Hollow fibers and Mammalian cells, Plant bodies, Interferons, Growth
membranes used, cells factors, Medicinal products
Rotorfermentor

 Modified Stirred Immobilized Bacteria, Ethanol, Monoclonal anti-


Tank Bioreactor Yeast, Plant cells bodies, Interferons, Growth
factors

 Modified Packed- Immobilized Bacteria, Ethanol, Enzymes, Medicinal


Bed Bioreactor Yeasts and other fungi products

 Tower and Loop Bacteria, Yeasts Single Cell Protein (SCP)


Bioreactors

________________________________________________________________________________________
Summary of Bioreactor Systems (Cont’d)
____________________________________________
Bioreactor Cell System used Products
design
_____________________________________________________________________________________________________________________

 Vacuum Bioreactors Bacteria, Yeasts, Fungi Ethanol, Volatile


products

 Cyclone Bioreactors Bacteria, Yeasts, Fungi Commodity products,


SCP

 Photochemical Photosynthetic bacteria, SCP, Algae, Medicinal


Bioreactors Algae, Cyano bacteria, plant products,
Plant Cell culture, r-DNA Monoclonal antibodies,
plant cells Vaccines, Interferons

________________________________________________________________________________________
SCALE-UP OF BIOREACTOR
SYSTEMS
DEFINITION OF SCALE-UP:

 Study of problems associated with the


transfer of experimental data from
laboratory and pilot-plant equipment to
large scale industrial equipment.
 The “ideal” scale-up criterion is that
parameter which has the same
numerical value as the volumes of the
geometrically similar bioreactors
increase in size.
HL3

HL2
HL1

DT1 DT2 DT3


LAB SCALE PILOT SCALE COMMERCIAL SCALE
FIG. 1 Geometric Scale-up of Bioreactors.

First scale-up criterion is the


 Preservation of Geometrical Similarity:
HL1/Dt1 = HL2/Dt2 =….. = HL3/Dt3…....(1)
Volume (VL)
 = Lab Scale Bioreactor VL1
 = Pilot Scale Bioreactor VL2
 = Large Scale Bioreactor VL3
EXAMPLE No.1 FOR SCALE-UP
For a given
 Medium Composition
 Temperature
 pH
We want to maximize the cell yield factor YX/S. We
start with a 10 L Laboratory scale bioreactor unit
and we perform optimization experiments at
different volumetric rates of oxygen supply, R.
Where:
R = KLa (CL* - CL)
= (moles O2)/(L)(hr)
YX/S = Cell to substrate yield
Factor based on glucose
= (g CDW yeast cells)/
(g glucose used)
Using the 10 L laboratory scale bioreactor
we carry out experiments and we get the
following hypothetical results shown in Fig.2
0.7
0.6
0.5 DATA FOR 10 L
BIOREACTOR
0.4
YX/S(-)

0.3
0.2
0.1
0.0
0 50 100 150 200 250 300 350
*
R = KLa (C L - C L)
FIG.2 10 L Bioreactor Hypothetical Fermentation Results for Yeast Aerobic Growth. Yeast
Cell Yield vs. Rate of Oxygen Transfer.

 QUESTION: When we scale-up to 50,000


L bioreactor system, are we going to get the
same YX/S vs. R relationship?
 It depends on what scale-up criteria we use
when we go from 10 L to 50,000 L
bioreactor systems.

 If the volumetric rate of oxygen transfer R


were a true scale-up criterion, then the
relationship between YX/S vs. R shown in Fig.2
for the 10 L bioreactor should be exactly the
same for any bioreactor size.
 For example, if experiments were done with
bioreactors of 10 L, 1,000 L, 10,000 L, 50,000
L or more, and the yield YX/S was measured
at different R, then the YX/S vs. R data for
each bioreactor volume should be the same
and independent of bioreactor volume.
 This would have been true only if R were a true
scale-up criterion. This hypothetical situation is
shown in Fig.3, where the data are the same for
different bioreactor volumes.
0.7
0.6
0.5 10 L BIOREACTOR
1,000 L BIOREACTOR
0.4 10,000 L BIOREACTOR
YX/S (-)

50,000 L BIOREACTOR
0.3
0.2
0.1
0.0
0 50 100 150 200 250 300 350
*
R = KLa (C L - C L)

FIG. 3 Hypothetical YX/S vs. R data for different bioreactor volumes, if R were an “ideal”
scale-up criterion.
 In reality scale-up of laboratory and pilot-
plant data to commercial size industrial
bioreactors is very difficult and complicated.

 No actual data or correlation exist for scale-


up.
 Different people use different scale-up
criteria to design commercial size bioreactor
systems.

 Also in industry there are a lot of trade


secrets on scale-up of bioreactors, and very
few published results exist in the literature.
n (r.p.m)

viscosity,
density
HL

Di

AIR, Q
DT
FIG.4 Independent Variables for a Bioreactor System.
 Scale-up criterion in general are a function of
independent variables n, Di, DT, HL, Qg, , .

 Several people used several different scale-up


criteria. Once a criteria is selected, then you
make sure that the numerical value of this
scale-up criterion is the same for the small
and large size bioreactor.
 Scale-up criterion small bioreactor = Scale-
up criterion large bioreactor.

 Example:
if we select the volumetric mass transfer
coefficient KLa to be the scale-up criterion,
then we make sure that when we scale-up we
have:
(KLa) small bioreactor = (KLa) large
bioreactor
Different Important DEPENDENT
variables used in scale-up.

1) Agitation Power, P or Pg

 From the Np vs. NRe figure, at


turbulent flow conditions:
Np = constant = Pgc/n3Di5
For a given fermentation broth of
density  and viscosity 

P  n3Di5…………………(2)
2) Rate of Liquid Pumping by the
Impeller, RL
 An Impeller serves a dual function of pumping
around of liquid inside the bioreactor vessel, and
local turbulent micromixing.

 Consider an impeller of diameter Di at a rotational


speed n (min-1). n

Di
 During one revolution the impeller “pumps”
around a liquid of volume VL, which is
proportional to the third power of the
impeller diameter Di, i.e. the liquid volume,
VL “swept” by the impeller.

VL  Di3
 Therefore the rate RL at which the impeller
pumps around the liquid is proportional to
n .VL

RL  n Di3……………………….(3)

Where: RL = (m3 liquid pumped)/(min).


 Therefore, if for a given rotational speed n,
the impeller diameter Di is increased by a
factor of two, then the liquid pumping rate
within the vessel, RL, will increase by a factor
of 23.
i.e. 8 times more.

 This demonstrates the powerful effect of


impeller diameter on the liquid pumping
rate, and how this affects the bulk mixing
within the bioreactor vessel.
3. Un-gassed Power Per Unit Liquid
Volume Delivered by the
Impeller (P/VL).
 For turbulent mixing conditions
according to Eq. 4.2:
P  n3 Di5
Also VL  Di3
Therefore,
P/VL  n3 Di2…………(4)
 For example, if the values of n and Di are
increased by a factor of 2 each, then the un-
gassed power per unit liquid volume is
increased by a factor of
(23)(22) = 32 more.

 This shows the powerful effect of increasing


both n and Di.
4. Impeller Tip Velocity, Vt

 The peripheral tip velocity, Vt, of the


impeller diameter Di, and the rotational
speed n (s-1), is given by:
Vt = (2R)(n) = (Di)(n)
Where: Vt = impeller tip velocity (m/s)

 The tip velocity of the impeller represents


the maximum velocity.
Vt  n Di…………………………(5)
 At the tip of the rotating impeller of diameter
Di we have maximum shear rate,  (s-1).

 Shear rate  = du/dx = (m/s)/(m) = s-1

and x is the distance away from the tip of the


impeller. In order to calculate the exact value of
shear rate, , we need to know the liquid velocity
profile distribution close to the impeller tip.
5. Impeller Reynolds Number, NRe.

 The Reynolds number based on impeller


diameter is given by:
NRe = n Di2/.

 Therefore, for a given fermentation broth of


given constant physical properties  and ,
we have:
NRe  n Di2…………………………….(6)
In general, the choice of scale-up criterion depends on two
considerations:

a) Nature of the fermentation and morphology


of the microorganism.
 aerobic
 anaerobic
 fungi
 single cell microorganisms
 mammalian cells
 plant cells
 exothermic fermentation
 Thermophilic microorganisms
 viscous or non-viscous fermentation broth
 Newtonian or non-Newtonian broth
b) During scale-up, what is the objective
parameter of fermentation we wish to
optimize (maximize).

 yield of product or biomass


 cell concentration
 product concentration
 product activity
 volumetric bioreactor productivity