IMPACTS OF BIOCHEMICAL ENGINEERING
IN BIOTECHNOLOGY
Kaushik Nath and Debabrata Das
Department of Biotechnology, IIT Kharagpur
B
iological systems are very
complex and beautifully
constructed, but they obey
the rules of chemistry and
physics and hence they are very much
susceptible to engineering analysis.
Many words have been used to
describe engineers working with
biotechnology. Biochemical
engineering, one of the most prolific
branches of biotechnology, usually
implies the extended application of
chemical engineering principles to
design, develop and analyze processes
using biocatalysts/ living organism.
These processes may result not only
in the formation of desirable
compounds but also in the destruction
of unwanted or hazardous substances.
Although it encompasses various
aspects of biochemistry, cell and
molecular biology, bioorganic and
bioinorganic chemistry, but however
has at its core the discipline of
chemical engineering. Biochemical
engineering is not only restricted to
well-defined artificially constructed
process; but also can be applied to
natural systems as well.
The practice of biochemical
engineering has a long history,
including such early applications as
the wine fermentation from grapes,
brewing of beer, cheese manufacture,
leavening of bread and effluent
disposal. In more recent times,
biochemical engineers have been
involved in antibiotic fermentation,
production of industrial solvents,
organic acids, vaccines, blood and
tissue products, animal feed stuffs,
commercial enzymes and in the
treatment of waste water/ effluents.
The various aspects of advancement
of biochemical engineering have been
summarized below.
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Bioreactor design and analysis
The bioreactor is the heart of all
biochemical processing. These are
vessels in which raw materials are
biologically converted into specific
products, using microorganisms, plant
or animal cells or individual enzymes.
A bioreactor supports the natural
process of cells trying to maintain their
environment to provide optimum
growth conditions - by providing
suitable conditions temperature, pH,
substrate, salts, vitamins and oxygen.
By combining our knowledge of
kinetics of biological reactions with
material and energy balances we can,
at least in principle design and analyze
the behavior of the bioreactor. Most
biological reactors are multiphase
systems.
Productivity, i.e. the amount of
product formed per unit of time is the
basis of any process design. To reach
maximum attainable productivity
within shorter period is a key factor in
any fermentation plant. Many
fermentation processes can be delayed
in reaching their maximum productive
capacities because of contamination
problems. Perhaps the most notable
contribution the engineer can render
is the design and operation of a
‘contaminant proof’ fermentation
vessel and its associated network of
piping. Firstly, sterility of the
equipment and the fermentation
medium has to be achieved. Next,
inoculum has to be passed in to the
fermenter without contaminating it.
The process then has to be maintained
in a pure state by preventing the entry
of contaminating organisms during the
fermentation. To accomplish these
objectives, the engineer changes
methods of vessel fabrication; revised
gasketing, piping and valve design and
also devises new methods of steam-
sealing possible points of contaminant
entry. In addition to designing an
aseptic fermentation operation, the
engineer also designs air-compression
and delivery system and efficient
methods for agitating and aerating the
fermentor. Provisions must be there to
remove heat evolved in metabolic
reactions within a narrow range. A
schematic diagram of a bioreactor is
given in Fig. 1.
Fig. 1 Schematic diagram of a
Bioreactor
The size of the fermenter
vessel is dependent on the
consideration of total projected
fermentation capacity. However there
is a greater risk of costly losses when
a large fermenter is contaminated.
When antibiotic fermentation first
became popular, S.S vessels (40-60
m3) were ubiquitous. Today, as the
technique of managing fermentation
aseptically improves, there is a
tendency to have larger and larger
fermenter vessels. Recent practice of
antibiotic and similar fermentations is
to have a batch extended with
repeated draw-offs, semi-continuous
and even continuous operations.
Because of increased productivity of
these units compared with strict batch,
the need for large vessels may decrease
and smaller vessels, 20-40m3 in size
may again become popular.
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Some practical problems with reactor
operation
a) Lack of homogeneity in a
continuous reactor: The cause of
poorer yields at low dilution rates
may be due to lack of
homogeneity in the reactor. It
comes more serious at low
dilution rates where the limiting
substrate concentration is low,
especially in case of viscous
media. To mitigate the problem,
good mixing of broth
mechanically and by aeration
becomes significant. Convenient
and reliable techniques are
needed for measuring the DO in
fermentation broth and for
following changes in viscosity
with time.
b) Gas hold up: Hold up of air
bubble in water filled agitated
vessels pose another problem in
reactor operation. This may cause
reduction of working volume of
the reactor
c) Maintenance of sterility: Even
with the most appropriately
designed fibrous air filter there
exists a finite chance of
contaminating microbes
eventually passing through the
filter in to the medium. To cope
with such eventualities, in
continuous glutamic acid
Table 1: Classification of Biochemical Reactors
Enzyme Biocatalysts
Homogeneous,
Liquid phase
Two-phase
(Gas-liquid)
Immobilized
biocatalysts
Entrapped biocatalysts
BSTR: Batch stirred-tank reactor; CSTR: continuous stirred-tank reactor.
14
fermentation the use of a mixture
of drugs which suppressed the
growth undesirable organisms
without hampering the actual
strain has been proposed.
d) Stability: There are two kinds of
stability involved; one concerned
with the microbial strain, the
other with mechanical
operations. Operation at dilution
rates near ‘wash out’ is not
recommended due to the
instability experienced in
practical operations. Knowledge
of ways to suppress undesirable
mutations or maintain the desired
strain in continuous cultivation is
needed.
More recent advances of
bioreactor designs include loop and
air lift fermenter, packed or fluidized
bed reactors using biocatalysts, and
hollow fiber and other membrane
arrangements for confining enzymes
or whole cells. A general classification
of different bioreactors is given in
Table 1. The airlift fermenter uses air
to agitate the fluid by means of draft
tubes or external recycle so that
expensive mechanical mixers can be
avoided. One very good application
for air-lift fermentation is the
production of single-cell protein with
bacterial cultures that are not very
viscous. Fluidized and packed bed
Microbial Biocatalysts
Aerobic
BSTR
CSTR — CSTR
BSTR
— CSTR
Bubble column
Airlift (loop)
Hollow fiber Airlift (loop)
Packed bed Packed bed
Fluidized bed Fluidized bed
BSTR
CSTR
Hollow fiber — Hollow fiber
Membrane
reactors usually use biocatalysts
encapsulated within a polymeric
carrier or fixed on to a solid or porous
support by a variety of methods, for
example covalent bonding, adsorption
and cross linking.
The basic goal of production at
minimum cost usually faces
constraints such as limited biocatalytic
capacity or upper bounds in
momentum, mass, and heat transfer.
Modern design objectives are:
• Cheaper raw materials as
substrate.
• High product concentration.
• High yield and fast conversion of
feedstock.
• Minimal by-product formation.
• Reactor flexibility for more than
one process.
• Affordable aseptic techniques.
• Minimum capital investment and
low maintenance.
• Effective control, that is, steady
product quality and quantity.
• Easy separation of organisms from
product stream.
• Minimal environmental impact.
Scaling up
Scale up studies constitutes another
important area where biochemical
Anaerobic
BSTR
—
Hollow fiber
Packed bed
Fluidized bed
BSTR
CSTR
Membrane
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engineering plays an important role.
The ultimate success of any chemical/
biochemical process lies in its
successful demonstration in pilot-plant
equipment and its subsequent scale-
up to the production stage. Scale-up
is the study of the problem associated
with transferring data obtained in
laboratory and pilot-plant to industrial
production. Equipment for the
fermentation industry’s fermentors,
heat exchangers, crystallizers,
separators and so forth, can be
designed and operated properly only
if scale-up technology is fully
understood.
Data analysis and parameter
estimation
A recent development in biochemical
engineering is the application of
computers in controlling the course of
bioconversion processes or
monitoring biochemical experiments
and estimating the reactor state from
real time measurements. In control
applications the most common
practice has been on-off control of
physical variables of the biochemical
process environment, for example
temperature, flow rates and foam. This
has been feasible because quick-
response sensors are generally
available for monitoring these
variables in real time.
By comparison, computer
control applications involving the use
of feed-back loops based on
measurements of the chemical
environments are less widespread,
although such methods are currently
under study. The reason for this is the
general unavailability of quick-
response sensors of the chemical
environment of biochemical
processes. Among the chemical
variables that are currently measurable
in real time are oxygen and carbon
dioxide concentrations in the effluent
and influent gas streams of aerobic
biochemical reactors, dissolved
oxygen concentration and pH.
The status of computer control may
be summarized as follows:
• The availability of a large number
of quick-response sensors (in real
time) for the chemical-biological
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environment would greatly
expand applicability. Biosensors
may be used to detect the
component in the micro levels.
• Current state and parameter
estimation procedures have been
applied successfully only to non-
product forming biochemical
reactions and stirred tank reactors;
there is the need to extend the
method to more general
biosynthetic reaction and
biochemical reactors.
• Computer control is a down
stream operation in the sense that
it requires the existence of an
actual biochemical reactor with
the computer and its associated
data acquisition paraphernalia. It
can only complement current off-
line reactor modeling that is very
much needed for design, scale-up
studies and the evaluation of
process economics.
Challenges and Opportunities in
Product Recovery
In down stream processes for product
recovery transport phenomena play a
decisive role. Isolation and recovery
of fermentation products are as
important as and sometimes more
expensive than the fermentation itself.
Driven by economic considerations
and the experience in the 1960s to
produce single cell proteins,
continuous recycle fermentation
systems are being developed to
replace the usual batch fermentor.
Higher concentrations of cells and/or
products and the desire to recycle the
cells without damaging them have
introduced the need for new recovery
processes. Other considerations
include acceptable product purity,
volume of production and stability of
the product to temperature, pressure,
ionic strength, pH and fluid shear.
Along with recent spectacular
advances in molecular biology has
been the development of a wide range
of new fractionation and
concentration technologies. Besides
improvements in the design and
operation of traditional separations
such as centrifugation, filtration,
adsorption, ion exchange and even
sedimentation, recent advances in
relatively new processes such as
membrane separations, chromato-
graphy, and electrophoretic
separations have expanded the
options enormously.
Many antibiotics have excellent
solubilities in organic solvents, which
are nearly water immiscible. A multi-
step, alternating aqueous to organic,
then organic-to-aqueous extraction
can provide both concentration and
subsequent purification. Operation of
the extraction step in penicillin as a
continuous process is accomplished
with a continuous flow, counter
current device. The light (solvent) and
heavy (broth) liquids enter through the
rotating shaft, and leave the rotating
contactor near the shaft and rotor
periphery, respectively. Rapid rotation
produces a centrifugal field, which
drives the two fluids counter current
to each other. A complete process flow
diagram of penicillin production along
with its purification in down stream
processing is presented in Fig. 2
Isolation of biochemical begins
with an extract of plant or animal
tissues or with a fermentation broth. If
the product has been released to the
medium, cells, debris, and various
solids are removed by filtration,
sedimentation, or centrifugation.
Products that are retained inside the
cells must first be released, usually by
rupturing the cells. This is followed by
clarification usually using
centrifugation. Recently the use of
membranes in place of centrifugation
for both cell separation and
clarification has been analyzed.
Different recovery and separation
techniques are listed in the following
Table 2:
Pollution abatement
In view of stringent environmental
regulations and growing public
awareness, industrial houses are
adopting elaborate measures to check
the pollution levels of the effluents,
they discharge. In the fermentation
industry, the treatment of wastes
should be regarded as an integral part
of plant design. If basic biochemical
engineering principles are applied to
the treatment of wastes before they are
diluted in a general waste water
15
 Fig. 2 Penicillin production process

(Ref. B. Atkinson and F. Mavituna, Biochemical Engineering and Biotechnology Handbook, Macmillan Publishers
Ltd., Surrey, England, 1983, p.990)

system, it would be expected that
costs would be considerably less than
those calculated from the cost of BOD
removal. Anaerobic treatment of the
final effluent proves suitable to
decrease nitrogen load by
denitrification.. Ions can be removed
by adsorption on resins or by reverse
osmosis, but these processes are
somewhat expensive. Trickling bed
filtration, activated sludge process are
the other useful techniques for waste
remediation. Moreover biochemical
engineering knowledge can make a
contribution to the management of
large bodies of water by attempting
to analyze the relationships between
nutrients and the growth they support
and between prey and predator along
the food chain. From such analyses, it
may be possible to predict when
instabilities are likely to be induced
in the system and to indicate what
conditions lead to stability.
Conclusion
Biochemical engineering has immense
possibilities. While the laboratory
scientists – the microbiologists, the
biochemists and the microbial
geneticist continue to discover and
advance desirable interactions
between micro-organisms and their
environment, the biochemical
engineer must control these
interactions and translate laboratory
results to production-scale operation
in an economic manner.
Improvements in management, as well
as in equipment, are needed to
accomplish these aims. In the long
run, it can revolutionize medical,
agricultural, and industrial technology.

Table 2: Recovery and separation Techniques

Precipitation Differential migration Filtration Sorption and partition Electrically driven

Coagulation and Sedimentation Deep bed Adsorption Ion Electrophoresis
flocculation Centrifugation Cake exchange Affinity Electrofocussing
Flotation Ultra filtration Gel filtration Free-flow electrophoresis
Hyper filtration Liquid-liquid partition Electro dialysis

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