Interdisciplinary Approach to Bioprocessing

Before the penicillin process, almost no chemical engineers required specialized training in
the life sciences. Even, until the time of Louis Pasteur in 1857, alcoholic fermentation was
considered to be a chemical process in which the sugars were getting converted to alcohol.
Subsequently, it was realized that the conversion was affected by microorganisms which
were the silent workers. This has opened up a wide range of interrelationships between
various disciplines in science. With the advent of modern antibiotics, the concept of a bioprocess engineer was born. The penicillin process also established a paradigm for bio-process
development and biochemical engineering.
The recent advances in genetic engineering, which help genetically engineer a cell to produce
large amounts of recombinant proteins
metabolites other than the proteins
have opened up new vistas of mathematical modelling of the cell as a micro batch reactor.
The cell can be forced, by applying genetic engineering principles, to produce high amounts
of the limiting enzymes. On the contrary, it may disrupt the cell metabolic process which may
ultimately lead to the cell death. Hence, to optimize the production, the culture has to be
maintained and forced to produce the desired enzymes without being perished in the process.
An advanced knowledge of cell growth and cell morphology, which are the subject matters of
microbiology, biochemistry and cell physiology, is a useful tool for precisely controlling the
biotechnological process. Modern biotechnological techniques such as recombinant DNA
techniques, gene manipulation, cell fusion and tissue culture offer useful tools to improve the
existing processes. Sophisticated medical kits, cultured human tissues, biopesticides,
microbial leaching of metals to reduce environmental pollution are some of the future visions
for biotechnological industry.
A successful scale-up technique can transform a scientific invention into a big
commercial reality. Since the behaviour of biological systems is always complex, their
scale-up is not easy. Hence, a rigorous evaluation and monitoring of various process

parameters in sterile environmental conditions can alone result in realizing the commercial
reality of the bioprocess. Application of mathematical modelling principles backed by
adequate pilot plant information will help in commercialization of the process. Thus, the
application of engineering principles to the biological processes to achieve commercial
success is the subject matter of biochemical engineering.

Biologists And Engineers Differ In Their Approach To Research:

The fundamental trainings of biologists and engineers are distinctly different. In the
development of knowledge in the life sciences, unlike chemistry and physics, mathematical
theories and quantitative methods (except statistics) have played a secondary role. Most
progress has been due to improvements in experimental tools. Results are qualitative and
descriptive models are formulated and tested. Consequently, biologists often have incomplete
backgrounds in mathematics but are very strong with respect to laboratory tools and, more
importantly, with respect to the interpretation of laboratory data from complex systems.
Engineers usually possess a very good background in the physical and mathematical sciences.
Often a theory leads to mathematical formulations, and the validity of the theory is tested by
comparing predicted responses to those in experiments. Quantitative models and approaches
even to complex systems, are strengths.
Biologists are usually better at the formation of testable hypotheses, experimental design, and
data interpretation from complex systems. Engineers are typically unfamiliar with the
experimental techniques and strategies used by life scientists.
The skills of the engineer and life scientist are complementary. To convert the promises of
molecular biology into new processes to make new products requires the integration of these
skills. To function at this level, the engineer needs a solid understanding of biology and its
experimental tools.

Outlines of Integrated Bioprocess

Integrated bioprocessing consists of various steps, some of them are sequential and others are
concurrent. They are shown in the figure below.

Figure 2: Integrated Bioprocessing Steps

Step 1: Isolation of the strain

The very first Step is identification and isolation of the strain of the microorganism which
brings out the desired bioconversion. This is generally the task of a microbiologist. This basic
work which involves some shake flask experimentation, etc., will be the basis for taking up
any process. The techno-economic viability of the process will be taken up subsequently if
the results of Step 1 are encouraging.

Step 2: Preservation of the strain

Preservation of the strain for future use is the basic requirement for the success of
commercial ventures. The high-yielding strains, identified and isolated in Step 1, will be
preserved for future use. It is essential that the strain is not only preserved, but its ability to
perform and yield the products should also be preserved, without which the whole exercise is
set at naught. The strains can be stored:
(i)

at refrigerated temperatures ( 2 to 6 C)

(ii)

at frozen temperatures (~18 to -80 C)

(iii)

by freeze drying (lyophilization)

Step 3: Growth of inoculums

Before using the strain in the fermenter, the inoculum is cultured for growth. The preserved
strain/culture is revived by growth in shake flasks or by solid-state fermentation on the solidsurfaces.
The standard growth time for lyophilized cultures is approximately 410 days, whereas the
frozen cultures and refrigerated cultures take different times for different cultures, as shown
in Table 1.
Table 1: Growth time for frozen cultures and refrigerated cultures
Cultures

Growth Time
Frozen culture

Refrigerated culture

Bacteria

4-48 h

4-24 h

Actinomycetes

1-5 days

1-3 days

Fungi

1-7 days

1-5 days

Step 4: Prefermentation culturing

Before the cells are admitted into the fermenter for performing bioconversions, they are
initially grown separately with nutrients and the substrate so that the cells can multiply and

proliferate. This will help increase the cell density. The optimal cell concentrations (%) in a
fermenter are as follows:
Bacteria : 0.1~3.0
Actinomycetes : 5-10
Fungi : 5-10
Step 5: Fermentation
The major activity in industrial operation in the whole process is fermentation, in which
biochemical engineers play an active role. Fermentation is the heart of bioprocessing
operation. The fermenter sizes vary from 1 to 450 m3, depending upon the type of
fermentation process. Based on the capacity, tonnage and nature of the fermentation product,
it could be taken up in: (i) batch fermenter (ii) fedbatch fermenter (iii) continuous
fermenter. The conditions of fermentation could be aerobic (with bubbling of air) or
anaerobic (in the absence of air)

Step 6: Recovery and purification of the product

The recovery and purification of the fermented product from the fermenter are crucial steps,
and sometimes even decide the economic viability of the process. The cost of recovery can
vary anywhere between 20 and 60% of the total manufacturing costs. Various stages of
operations involved in this step are shown in Fig 2. The methodology of primary isolation of
the product depends upon the nature of the cells and how they hold the product, whether it is
intracellular (the product is held within the cells) or extracellular (the product is excreted
from the cells).
Any solids, etc. are removed by simple filtration or centrifugation techniques. The
intracellular material is obtained by cell disruption techniques. Some of these techniques
which seem to be simple and feasible at laboratory scale, may not be so at the commercial
scale, particularly for intracellular enzyme extractions.

For purification and isolation of the final product, different unit operations could be
employed as shown in Figure 2.

Step 7: Treatment of effluent

Ferrnentation processes release large amounts of effluents which may be rich in organic
matter, and may be a potential hazard for the environment if let out as they are. They need to
be treated before being released into the outside world. The ability to economically treat the
effluents before they are let out into the environment makes or mars the viability of the
product manufacture. The various effluent treatment techniques can be classified as:
(i)

Physical

(ii)

Chemical

(iii) Biological
The final choice depends upon the individual cases and local circumstances.

Traditional and Modern Applications of Biotechnology