1

INTRODUCTION TO BIOCHEMICAL ENGINEERING

BIOTECHNOLOGY
Recombinant DNA allows the direct manipulation of genetic material of individual cells, which may be used to develop
microorganisms that produce new products as well as useful organisms. A major objective of this technique is to splice a
foreign gene for a desired product into circular forms of DNA (plasmids), and then to insert them into an organism, so
that the foreign gene can be expressed to produce the product from the organism.

Cell fusion is a process to form a single hybrid cell with nuclei and cytoplasm from two different types of cells in order to
combine the desirable characteristics of the two.
Applications of Biochemical Engineering
Pharmacogenomics- Gene Inheritance and response to drugs
Genetic testing- Direct examination of DNA
Gene therapy- Curing genetic and acquired diseases like AIDS and cancer
Human Genome Project- reference sequence for the entire human genome and identify all the human genes
Cloning- the removal of the nucleus from one cell and its placement in an unfertilized egg cell whose nucleus has
either been deactivated or removed.

BIOLOGICAL PROCESSES
Raw materials, usually biomass, are treated and mixed with other ingredients that are required for cells to grow well.
The liquid mixture, the medium, is sterilized to eliminate all other living microorganisms and introduced to a large
cylindrical vessel, bioreactor or fermenter, typically equipped with agitators, baffles, air spargers, and various sensing
devices for the control of conditions. A pure strain of microorganisms is introduced into the vessel. The number of cells
will start to multiply exponentially after a certain period of time and reach a maximum cell concentration as the medium
is depleted. The process will be stopped and the contents will be pumped out for the product recovery and purification.
This process can be operated either by batch or continuously.

PROCESS DESIGN & DEVELOPMENT
1- One must have an understanding of the basic sciences for the process involved. These are microbiology,
biochemistry, molecular biology, genetics, and so on.

2- Kinetics deals with rate of a reaction and how it is affected by various chemical and physical conditions. This is where
the expertise of chemical engineers familiar with chemical kinetics and reactor design plays a major role. To design
an effective bioreactor for the biological catalyst to perform, it is also important to know how the rate of the
reaction is influenced by various operating conditions. This involves the study of thermodynamics, transport
phenomena, biological interactions, clonal stability, and so on.

3- For the optimum operation and control, reliable on-line sensing devices need to be developed. On-line optimization
algorithms need to be developed and used to enhance the operability of bioprocess and to ensure that these
processes are operated at the most economical points.

4- The downstream processing (or bio-separation), a biochemical engineer can utilize various separation techniques
developed in chemical processes such as distillation, absorption, extraction, adsorption, drying, filtration,
precipitation, and leaching. In addition to these standard separation techniques, the biochemical engineer needs to
develop novel techniques which are suitable to separate the biological materials

2

CHARACTERISTICS OF BIOLOGICAL PROCESSES
Specificity: An enzyme catalyst is highly specific and catalyzes only one or a small number of chemical reactions.
A great variety of enzymes exist that can catalyze a very wide range of reactions.
Effectiveness: The rate of an enzyme-catalyzed reaction is usually much faster than that of the same reaction
when directed by nonbiological catalysts. A small amount of enzyme is required to produce the desired effect.
Recombinant DNA technology: The development of the recombinant DNA technology promises enormous
possibilities to improve biological processes.
Variability: Cells tend to mutate due to the changing environment and may lose some characteristics vital for the
success of process. Enzymes are comparatively sensitive or unstable molecules and require care in their use.

MICROBIAL GROWTH CURVE
Lag phase is due to physiochemical equilibrium between microorganism & environment following inoculation. It
can be time consuming and costly so it should be shortened. This can be achieved by growing the inoculum in
comparable medium to bioreactor and under similar growth conditions (pH, T) i.e exposing it to minimum stress
level. A minimum of 5% (v/v) inoculum of exponentially growing cells should also be used for inoculation.
Once the cells have adapted to the new conditions of growth, they enter the exponential phase.
After the deceleration phase, growth ceases and the culture enters a stationary phase or a steady state. The
biomass remains constant, except when certain accumulated chemicals in the culture lyse the cells (chemolysis).
Unless other micro-organisms contaminate the culture, the chemical constitution remains unchanged.
Mutation of the organism in the culture can also be a source of contamination, called internal contamination.
Eventually, the cells enter death phase and this is characterized by a drop in optical density and biomass levels in
most cultures.

The batch culture growth curve gives a good indication of when to stop the fermentation. The fermentation can be
terminated at the end of the exponential growth phase before the cell enters stationary phase.

FERMENTATION PROCESS
All fermentation processes involve inoculation of a selected and specially bred strain of microbe, plant or animal cell into
a sterile medium held within a sterile fermenter vessel. After medium sterilization, the organism is inoculated into the
vessel and allowed to grow.

FERMENTER TYPES
1. Stirred tank Reactor (SRT)
Most Common Type
Aspect Ratio: 3:1
Can be operated in Batch or continuous mode
2. Tower Fermenter
Aspect Ratio: 6:1 to 15:1
Continuous Operation
3. Air Lift Fermenter
Mixing system is replaced by a constant flow of gas introduced into a riser tube
Baffled to improve mixing
Suitable for Shear sensitive cells
4. Hollow Fiber Chamber
Used to grow anchorage-dependent cells
System consists of fiber bundles, and the cells grow within the capillary space within the cartridge
5. Micro-carriers
Chromatographic grade DEAE (diethylaminoethyl) positively charged beads
Attracted to negatively charged animal cells and provide a surface on which the cells can grow
3

LAYOUT OF FERMENTATION FACILTY
The left hand side shows the layout in a separate air-conditioned room containing the steam generators, air
compressors, gas bottles and storage. This room is noisy and hot, and is separated from the main fermenter room,
illustrated on the right of the diagram, by a corridor. Pipework through the ceiling space above the corridor connects the
two rooms and a set of utilities is delivered to each of the fermenters through appropriate power outlets, valves, and
taps. The air-conditioned room on the right contains the three fermenters, their purified water feeds, the centrifuge, a
fume hood, a dry bench for the computers, and a wet bench with sink for cleaning and washing equipment. Most of the
preparation of media and autoclaving of glassware and solutions is carried out in the main lab to the right of the
fermenter room.