ECH4202

TOPIC 1
INTRODUCTION TO
BIOREACTOR
Semester: II- 2018/2019
Lecturer: Assoc. Prof. Dr. N Abdullah
(email:[email protected]; hp: 0122480122)
 ECH4202

Overview of bioprocess industry

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

 ECH4202

Learning objective

1. To evaluate basic design component of bioreactor

2. To assess the function and application of different types of
bioreactor
 ECH4202

Content
1. What is bioreactor & its Design Criteria
2. Component of bioreactors
3. Types of bioreactor configuration commonly used in
industry
4. Example of Novel Bioreactor Types
 ECH4202

1. What is a Bioreactor?
• A vessel to contain biological reaction
• supply a homogeneous environment for its content.
• Provide controlled environment
• For growing organisms (yeast, bacteria, or animal cells) under
controlled conditions.
• For industrial processes to produce pharmaceuticals, vaccines, or
antibodies
• For convert raw materials into byproducts
 ECH4202

Typical bioreactor set-up

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)
 ECH4202

Basic Bioreactor Design Criteria

1. Microbiological and Biochemical Characteristics of the Cell
System (Microbial, Mammalian, Plant)
2. Hydrodynamic Characteristics of the bioreactor
3. Mass and Heat Transfer Characteristics of the Bioreactor
4. Kinetics of the Cell Growth and Product Formation
5. Genetic Stability Characteristics of the Cell System
6. Aseptic Equipment Design
7. Control of Bioreactor Environment (both macro- & micro-
environment)
8. Implications of Bioreactor Design on Downstream Products
Separation
9. Capital and Operating Costs of the Bioreactor
10. Potential for Bioreactor Scale-up
 ECH4202

2. Bioreactor Components
• Vessel • Cooling Jacket
• Agitator • Ports for input and
• Sparger output of material
• Baffles • Condenser
• Probes • Back Pressure Valve
 Temperature • Inlet Filters and
 Dissolved Oxygen
Exhaust Filters
• Valves
 pH
• Load Cell
 Pressure Gauge
 ECH4202

Vessel (Continue)
• Function: hold media, cells
• Materials: glass, SS, durable plastic
• Other parts (ports, accessory, auxiliary attached to it)
 ECH4202

Vessel

Head Plate

Vessel
 ECH4202

Agitator
• For mixing A
• consist of a shaft and impellers.
• Mixing :
• to supply nutrients and oxygen to the culture
• to maintain a constant pH and temperature.
• Impellers come in many different design.
 ECH4202

Agitator

Agitator Shaft

Impellers
 ECH4202

Sparger
• to introduce gasses into the
vessel.
• located at the bottom of the
vessel
Micro Sparger
• consist of a tube with tiny holes
• helps to aerate, supply oxygen &
mix the contents
Openings on Ring
Ring Sparger Sparger
 ECH4202

Baffles
• obstructions on the side of the vessel that generate
turbulence in the flow of the culture.

Baffle
 ECH4202

Probes
• Bioreactors require probes to monitor the culture in the
vessel.
• Eg: probes for temperature, pH, DO (dissolved oxygen),
and CO2
 ECH4202

Probes

Probe Belt
Probes on head plate

Probes
 ECH4202

Cooling Jacket
• To maintain a constant
temperature
• Coolant (cold water or glycol)
flows through the cooling
jacket to regulate the
temperature.

Stainless Steel Cooling Jacket

surrounds the vessel.
 ECH4202

Ports
• Ports for material to be introduced or removed from the
vessel.
• Example: media port, inoculation port, nutrients (feed
ports), pH control port (acid/alkaline), sample port
 ECH4202

Bioreactor - Ports

Ports
 ECH4202

Condenser
• An apparatus that captures
saturated air leaving the vessel.
• Also help to minimize the loss of
material out of the reactor due
to evaporation.
 ECH4202

Filters
• inlet filters: to ensure the
gasses entering the vessel
are sterile.
• exhaust filters: to keep the
reactor sterile and allow gas Filter Housing
to escape to regulate
pressure.
• Filters require a filter
housing – a stainless steel Filter
cabinet to hold and sterilize
the filter.
 ECH4202

Back Pressure Valve

• pressure in the reactor to be higher than the pressure surrounding it
• Constant pressure helps to ensure sterility of the reactor
• The pressure is regulated by : a back pressure valve.
 ECH4202

Valves

• to control the flow of either gas or liquid

• Bioreactors use many different types of valves.
• Manual Valves
• Pneumatic Valves
Pneumatic (Automatic)
• Steam Lock Valves Valve
• Used at different location in bioreactor
• Sample valve
• Harvest valve
• ports

Steam Lock Valves

Manual Valve
 ECH4202

Load Cell
• a scale that measures the weight of bioreactor.
• Bioreactor sits on a series of load cells that compress
when weight is added to the vessel.
Weight = Density x Volume
 ECH4202

Load Cell

Compact
Compression
Load Cells

Spacing of
Load Cells
 ECH4202

Utilities
• Bioreactors require utilities.
• Gasses
• Air
• Oxygen
• Carbon Dioxide
• Nitrogen
• Clean Steam - for sterilization of the reactor
(condensed clean steam equals WFI (water for
injection) that is clean enough to be injected)
• Coolant (cold water or glycol) – for cooling jacket
• Instrument Air - to operate pneumatic valves
 ECH4202

3. Types of bioreactor configuration

commonly used in industry

a) Stirred tank
b) Bubble column
c) Air lift
d) Fluidised bed
e) Packed bed
 ECH4202

a) STR
• Mechanical stirrers are used for mixing, distribute heat and
materials (O2 & substrates)
• More than 70% industrial work use this
• Not the best. Batch STR generally have low volumetric
productivity
• Downtimes are large
• Unsteady state causing microbes to stress, limit to nutrient
• Fed batch may eliminate batch problem
• Cost of operation is lower & can be used for many type of
microbial species
• Not recommended to run with immobilised cell
 ECH4202

STR
 ECH4202

b) Bubble column
• Tall reactor, use air alone to mix the contents
 ECH4202

c) Air lift
• Similar to bubble column, but differ by having a draft tube.
• Pneumatic reactors (no stirrer, use expansion of compressed air)
• Air/liquid introduced upward motion
• Draft tube: inner tube to improve circulation & oxygen transfer
& equalize shear forces
• Can be used for both free and immobilised cells
 ECH4202

Airlift reactor
 ECH4202

d) Fluidised Bed
• Cells are ‘immobilised’ as
small particles that move
with fluid
• Small particles create
large surface area for cell
to sticks to and high rate
of oxygen and nutrient
transfer
• Volumetric productivity
usually higher than STR
and packed bed
 ECH4202

Fluidized Bed Bioreactors (FBB)

 ECH4202

e) Packed Bed
• Cells are immobilised on large particles. Large particles don’t
move with liquid
• Simple to construct & operate
• Suffer from blockages and poor oxygen transfer, channeling
• Modification to improve pressure drop: incline bed, horizontal
bed, rotary horizontal bed- but limited success
• Used with attached biofilm wastewater engineering
• Immobilised biocatalyst- nutrient fed either top or down flow
 ECH4202

Packed Bed Bioreactors

Example of bioreactor & its application
__________________________________________________________
Bioreactor Cell Systems Products
Design used
__________________________________________________________
Air-Lift Bioreactor Bacteria, Yeast and SCP, Enzymes, 2o
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 mAb,
Vaccines, Proteases,
Hormones

__________________________________________________________
 Examples of Bioreactor & its application
________________________________________________________________________
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
Rotor fermenter

Modified Stirred Immobilized Bacteria,Ethanol, Monoclonal antibodies

Tank Bioreactor Yeast, Plant cells 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

_________________________________________________________________________________
Examples of Bioreactor & its application
_______________________________________________________________________________
Bioreactor Cell System used Products
design
_______________________________________________________________________________
Vacuum Bioreactors Bacteria, Yeasts, Fungi Ethanol, Volatile
products

Cyclone Bioreactors Bacteria, Yeasts, Fungi Commodity products,

SCP

l Photochemical Photosynthetic bacteria, SCP, Algae, Medicinal

Bioreactors Algae, Cyano bacteria, plant products, rDNA
Plant Cell culture, Monoclonal antibodies,
plant cells Vaccines, Interferons

______________________________________________________________________________
 ECH4202

4) Other Bioreactors: Novel Bioreactors

• Categories of novel bioreactors
• Modified STR
• Tubular bioreactor
• Immobilised cell
• Why need a novel bioreactors?
• Biological development
• Biotech yields many product which have special requirement and need
‘tailor-made’ reactor
• Biochemical Engineering development
• Retention time of cells, cell cycle
• Competition within fermentation industry
• Strain selection with novel ideas.
 ECH4202

Example of novel
bioreactor: a tower Air exhoust
bioreactor system with
perforated plates and co-
current air liquid flow. Broth
outlet

Sampling
nozzles
Jacket

Constant temp.
water bath Perforated
Pump
plate

Compressed
air Orifice
Sparger

Air filter
Peristaltic Medium
pump reservior
Flow
Medium meter
inlet
 ECH4202

Air Feed

Perforated
plate

Example of Novel Bioreactor:

a tower bioreactor system with
multiple impellers and liquid down Downcomer

comer and counter-current air

liquid flow
Baffle

Impeller

Product
Air
 ECH4202

AIR SLUDGE
INLET RECYCLE

OUTLET

Example of Novel
Bioreactor:
START
ICI Deep Shaft Unit -UP AIR RISER

PROCESS
AIR
DOWN-
COMER

SHAFT
LINING
 ECH4202

CONDENSATE,
MAE-UP WATER, AND
CLARIFIER FLOCCULATING AGENT
WASH
WATER
SETTLEMENT RECYCLED
TANT WATER
SAND B RECYCLE SLUDGE
B

DECANTER
CENTRIFUGE FLOATATION

Example novel SOIL AND

SLUDGE
Bioreactor: DEEP
Emlichhiem SHAFT

Chemical
AIR

COMPRESSOR

LAGOON
 ECH4202

Outer draft tube

Inner draft tube

1.176 2.620 4.530

4 Jets
Internal circulation patterns of fluidized
Ca-alginate beads containing
immobilized cells of Z. mobilis. All
dimensions in cm.

28.40

21.30 26.43

6.895
2.876

0.1
0.953
 ECH4202

Vacuum Fermenter Chilled

water

Condenser
Vacuum
Level pump
control
Vacuum
control
Receiving
tank
(product)

Dry ice
bath

Heating Fermenter
water
Filter

Medium
reservoir
Metering Receiving
Rheostat pump tank
Filter
(bleed)

Air or O2
 ECH4202

Individual assignment (10% mark)

Propose an innovative design of bioreactor for 1 selected bioproduct from
the list below:
1) Rheologically challenged product
2) Low solubility product (due to aggregation etc.)
3) Fermentation system with product inhibition (ie: lead to low yield)
4) Thermally sensitive product
5) ‘Sticky’ product (ie: hydrophobicity concern)

In your paper: state the design configuration to tailor made the culture
growth and product formation (include growth & product kinetics, mass
transfer, mixing, rheology, instrumentation etc) .
Also sketch the proposed bioreactor design (using design drawing tools
that you have used in FYDP course)

(Note: PO assessed are: PO1, PO3, PO9, PO14)