INTRODUCTION TO

BIOREACTOR DESIGN

EKC 351: BIOCHEMICAL ENGINEERING

ASSOC. PROF. DR. KHAIRIAH ABD KARIM
[email protected]
Course Learning Objective 2
Analyze various problems related to bioreactor
operations based on different model systems.

Course Learning Outcomes

By the end of this topic, student should be able:
o To compare between bioreactors and chemical
reactors.
o To describe the key issues in bioreactor design and
operation.
o To understand the requirement of operating a
bioreactor and bioreaction process.
Introduction
❑Selection and specific design of a bioreactor system
include a series of decisions on matters ranging
from basic microbiology and biochemistry to
process engineering and marketing.
❑All those disciplines must contribute in a harmonious
way to make possible proper determination of basic
type of bioreactor system and its correct
dimensions in order to provide an optimal
environment for the biological system, operating at
minimal cost, so that the maximum productivity can
be achieved and benefits of the process can be
obtained.
❑ Bioreactor design is a complex engineering task.

❑ Under optimum conditions, the microorganisms or cells

are able to perform their desired function with 100
percent rate of success.

❑ The bioreactor's environmental conditions like gas (i.e.,

air, oxygen, nitrogen, carbon dioxide) flow rates,
temperature, pH and dissolved oxygen levels, and
agitation speed/circulation rate need to be closely
monitored and controlled.

❑ Most industrial bioreactor manufacturers use vessels,

sensors and a control system networked together.
Figure 1: Typical process flow for production of bioproducts
Upstream Process

Downstream Process
Figure 2: Bio-ethanol process from starch. Adapted from Ochoa et. al. (2010), Journal of Process
Control Vol. 20(9): 983-998.
Product requirement
❑The potential products cover a wide range, from inexpensive,
large quantity products like ethanol, clean water or soil, to the
most sophisticated and complex biochemical products
(biopharmaceuticals), which are expensive and required in
small amount.
❑For inexpensive products – price is determined by equipment
and operation costs.
❑For expensive products – price is determined by costs
associated with legal approval by regulatory authorities (e.g.
Food and Drug Administration (FDA)) and the uniqueness of
product.
❑Therefore, once the target product has been specified using
biochemical route, then the system to be used must be
carefully chosen.
 Table 1: Typical bioprocess and their market volume
Product Annual volume Approximate Price ($/kg)
(metric tons) value ($billion)
Ethanol 19 000 000 5 0.25
Citric acid 1 100 000 1.1 1
Glutamic acid 800 000 0.8 1
Detergent protease 100 000 0.3 3
Aspartame 10 000 0.05 5
Cephalosporins 5 000 2.5 500
Tetracyclines 5 000 0.3 60
Insulin 8 1 125 000
Erythropoietin* 0.01 5 500 000 000

*A growth factor that stimulates the production of red blood cells

Bioreaction Products
Process 1
❖ Product is produced by cells, i.e. extracellular (alcohols or
citric acid) or intracellular (a metabolite or an enzyme).

❖ Two types of cellular products:

o Primary metabolites - produced during the growth
phase and are needed for growth). e.g. amino acids,
proteins.

o Secondary metabolites - produced from intermediates,

products of primary metabolism; maybe toxic and
possess antibiotic properties.
 Process 2
❖ During this type of process, a cell mass is produced.
❖ E.g. 1) Baker’s yeast: used in baking industry.
2) Single cell proteins: food sources

Process 3
❖ Involves modification of a compound that is added in the
fermentation process  BIOTRANSFORMATION
❖ Process involved: Dehydration, oxidation, hydroxylation,
amination etc.
❖ Products produced are steroids, antibiotics, hormones etc.
 Bioreactors vs Chemical Reactors
❑A bioreactor refers to “any device
or system that supports a
biologically active
environment”.
❑A bioreactor includes mechanical
vessels in which:
✓ Organisms are cultivated in
a controlled manner
✓ Specific reactions are
involved in converting or
transforming materials.
❑This process can either be
aerobic or anaerobic.
❑Bioreactors are specifically designed to influence the metabolic
pathways:
 CSTRs, PFRs, Fluidized Bed Reactors, Bubble Column Reactor
❑ Bioreactors differ from conventional chemical reactors in that
they support and control biological entities.

❑ As such, bioreactor systems must be designed to provide a

higher degree of control over process upsets and
contaminations, since the organisms are more sensitive
and less stable than chemicals.

❑ Biological organisms, by their nature, will mutate which may

alter the biochemistry of the bioreaction or the physical
properties of the organism.

❑ Although the majority of fundamental bioreactor engineering

and design issues are similar, maintaining the desired
biological activity and eliminating or minimizing
undesired activities often presents a greater challenge
than conventional chemical reactors.
❑ Biological factors to be considered in a typical bioreaction
process include the characteristics of the cells, their
maximum specific growth rate, Monod constant, yield
coefficient, pH range and temperature range.

❑ The productivity of a fermentation is determined by the mode

of operation of the fermentation process; e.g. the
advantages of fed-batch and continuous fermentations over
batch fermentations.

❑ A bioreactor is not an isolated unit, but is a part of an

integrated unit with both upstream (preparation and
downstream (recovery) unit operations.

❑ Bioreaction takes about 10-20 days for completion of a batch

process.
 Key Issues in Bioreactor Design and
Operation
❑ A bioreactor should comply with:
1) GMP – Good Manufacturing Practice, regulations
(cGMP – Current GMP)
2) Three Qs:
o Design Qualifications (DQ)
o Installation Qualifications (IQ)
o Operation Qualifications (OQ)
3) Validation – Established Documented Evidence
(purpose to be served).
4) cGMP – Employed strictly in drug industry, where
stringent requirements are met.
❑ To accomplish this, a chemical engineer must consider the
following two areas:
1) Suitable reactor parameters (for desired biological,
chemical, physical systems)
2) Bioreaction parameters:
o Controlled temperature
o Optimum pH
o Sufficient substrate (carbon source)
o H2O availability
o Salts for nutrition
o Vitamins
o Oxygen
o Gas evolution
o Product or by-product removal
 Components of A Typical Fermentation
Process

Figure 3: Components and operations of a typical fermentation process

 Process Function
Medium preparation o Nutrients in proportion to minerals are added in
(formulation) sufficient quantities for a specified amount of biomass
or product to be synthesized. Crude sources such as
beet, cane, corn, molasses can replace purified sugars.

Medium sterilization o Purpose: to avoid process upsets and contaminations.

o Steam or chemical addition accomplishes the medium
sterilization. It is an important component in
fermentation process.

Inoculum preparation o Comprises of 5-10% of new medium volume.

Air filtration o To remove the traces of various components that are
present in the gaseous form & to remove moisture.
o Filtered air is inducted through the sparger.
Process control o The overall monitoring of the process is controlled and
set.
Cell recovery o Downstream processing part, recovers all the valuables
present in the bioreactor.
Design and Operation of A Typical Aseptic
and Aerobic Fermentation Process

❑ Why is the fermentation

process to be aseptic?
- To avoid any kind of
interruptions, process
upsets or contaminations.

❑ Preparation of inoculum –
need careful development of
relatively a few cells to a
dense suspension (10 –
20% of bioreactor volume).

Figure 4: Valve and piping configurations

Upstream Process:
Sterile and aseptic condition
How to transport inoculum aseptically
from seed tank to the fermenter?

1) Install pipe section AB.

2) Sterilize connection with 15 psig
steam for 20 min. Valves D and J are
open and valve C is closed.
3) Cool the fermenter under sterile air
pressure with valves C, G, H, I and J
closed and valves D, E and F open.
Sterile medium fills the connection.
4) Increase pressure in the seed tank to
10 psig. Lower the fermenter
pressure to 2 psig.
5) Transfer the inoculum by opening
valve C.
6) Steam seal the fermenter-seed tank
connection by closing C and F and
opening G and J. Steam and
condensate are drained from partially
open D and E.
 Choosing the Cultivation Method
Batch Culture Continuous Culture
o Growth slower as nutrients o Growth faster as nutrients
decline always there
o Easy to set up and maintain o Difficult to set up and
maintain
o Only one batch lost if o Huge volumes lost if
contamination occurs contamination occurs
o Less efficient, fermenter not o More efficient fermenter in
always in use use constantly
o Useful for producing o Useful for producing primary
secondary metabolites metabolites
 Batch Culture Continuous Culture
o Fermentation in a closed or o Fermentation in an open
batch fermenter fermenter
o Microorganisms and nutrient o Nutrients added at a steady
medium added and left for a rate throughout the process
period of time
o Nothing added to or o Maintain microorganisms at
removed from the fermenter exponential phase of growth
during the process (except
venting of waste gases)
o Product separate at the end o Product removed at a steady
rate throughout the process
o Temperature controlled and o pH, temperature and oxygen
nutrients usually depleted at concentration as well as
the end nutrient and product levels
should be kept constant
 General Features of Bioreactor Design
❑ A bioreactor: an apparatus shaped like a
chamber for growing organisms such as bacteria
or yeasts, used for the production of biomolecular
metabolites or biopolymers or for the conversion of
organic wastes.

❑ The main purpose of the design efforts: to

accomplish conditions where diverse cell types
are able to grow efficiently and produce a
variety of biological products with a wide range
of molecular sizes in a single unit.

❑ Time factor: rates differ largely from one organism

to another, in reproduction rates, in rates of
molecular processing in the individual organisms,
and transfer across biological barriers of the
cellular systems.
❑ Operational procedures: When cells grow, the design must
adapt to compensate for the magnification of the dynamics due
to higher cell numbers.

❑ Inoculation of cells: relates to the size of the inoculum, the

state of the cells to enter an exponential growth phase, their
variability and sensitivity to micro-environmental conditions,
and their purity.

❑ Media composition: another design issue comprising both the

chemical aspects related to the nutritional value of the media
as well as the biological significance of the components in
relation to metabolic pathways involved in the growth and
production of the cells.

❑ Table 2 summarizes the most essential conditions and key

parameters and how they influence design work and
options.
Table 2: Bioreactor design criteria
Table 2: Bioreactor design criteria.. cont.
❖ Based on criteria listed in Table 2, a diversity of bioreactor
design alternatives have emerged:
i. Stirred tank bioreactor - predominantly used design
for submerged cultures.
ii. Bubble column bioreactor - mechanical impeller is
exchanged with raising bubbles
iii. Airlift bioreactor – a forced flow in an internal or
external loop
iv. Fluidized bed reactor - cells are recycled by external
pumping and soluble product harvested by overflow
v. Trickle-bed bioreactor - cells are grafted to a solid
material while the medium is fluxed through a bed of
biocatalyst
vi. Solid state bioreactor - follow the ancient Chinese
tray reactor model as applied in koji fermentation.
Conventional vs Novel Bioreactors
➢Conventional bioreactors – stirred-tank reactor (STR),
spinner flasks
➢Novel bioreactors – rock-bed-single-use bioreactor,
rotary cell culture system, air-lift single-use reactor,
fluidized-bed reactor
➢The main difference – functionality and design of the
bioreactor:
o Conventional bioreactors: use for simple
fermentation, design usually a typical one (stirrer and
sparger)
o Novel bioreactors: design more complex and suitable
with fermentation approaches using special types of
cell.
Conventional Bioreactors

Stirred tank bioreactor Spinner flask bioreactor

Novel Bioreactors

Bioreactor that
incorporates a LIGHT
SOURCE to provide
photonic-energy input
into the reactor.

Used for the cultivation

of photosynthesizing
organism (algae).

Photobioreactor
Bubble column bioreactor Airlift bioreactor

A tall column bioreactor where gas is Similar to bubble column bioreactors

introduced in the bottom section for but they contain a draft tube 
mixing and aeration purposes. improves circulation and oxygen
transfer and equalizes shear forces in
the reactor.
Wave Tidal Bioreactor -
disposable cell culture bioreactor.

This bioreactor consists of a

sterile wave bag and rocker. The
single-use bag can be eliminated
by any needs for cleaning or
sterilization.

The rocker can be set and control

temperature. Also, it makes
regular rocking motion that
induces waves in culture media.

Wave action makes cells

suspense and prevents
cumulating. Wave actions for
mixing and transferring oxygen
results in perfect environment for
cell growth.