includes general information about fermentation process.

1. FERMENTATION
Fermentation is a biochemical process employing selected micro-organisms
and microbiological technologies to produce a chemical product.

2. DESIGN OF THE FERMENTATION PROCESS
Any fermentation process design must consider
three major aspects:
(i) Value creation opportunity
(ii) Process design analysis
(iii) Objectives of the design project

3. I. VALUE CREATION OPPURTUNITY
It concerns mostly establishment of microbial
reaction process and its development recognizing
economic opportunity.
To create and deliver value in an efficient enough
way that it will generate profit after cost.
IMPORTANCE?
Because value creation is the starting point for all
businesses, successful or not, it's a fundamental
concept to understand.

4. Recognition Of Economic Opportunity
Relates to the Appropriateness Of:
1. SELECTION OF MICROBIAL STRAINS
2. SELECTION OF APPROPRIATE FERMENTER DESIGN
3. SCOPE OF PRODUCT UTILIZATION

5. Exploitation Of Microbial Activity Is Done
Considering:
Selection of new stable
strains for getting a
share in the market by
producing useful new
products using modern
fermentation plant.
Selection of stable strain
for known product of
the established
equipment.

6. DESIGN OF THE FERMENTER
For production of a substance having established market,
combination of an existing stable strain and a simplified fermenter
together with new devices need to be used.
In almost all fermentation processes the fermenter configuration
plays the part of a mediator between energy input variables and
physiological reactions of microbial cultures.
As microbial behaviour vary widely, it is difficult to design a
versatile fermenter for multipurpose use. This limitation led to
the development of various conventional and non-conventional
fermenter designs.

7. DESIREABLE REQUIREMENTS OF A FERMENTER

8. High power demand and underutilization of impeller energy
input are the two important weak points of conventional
designs.
In any new design, principles of unconventional fermenters
have been to minimize these shortcomings of conventional
fermenters.
Also, care is needed in the interaction between fermenter
design and microbial unit at used physiochemical environment
to make “the downstream processing of the product ”easier.
Downstream processing refers to the recovery and purification
of biosynthetic products, particularly pharmaceuticals, from
natural sources such as animal or plant tissue or fermentation
broth, including the recycling of salvageable components and
the proper treatment and disposal of waste.

9. INTEGRATION OF MICROBIAL UNIT WITH DOWNSTREAM
PROCESSING

10. SCOPE OF THE PRODUCT
It is associated with the “mode of utilization” of
the product to get an adequate fitness of the
product on human needs.

11. II. PROCESS DESIGN ANALYSIS
Fermentation time course and profiles of state and control
variables.
Determination of most favorable control variables condition to
obtain maximum product yield.
Pilot plant studies for new process development thereby aid in
innovation in scale up.
Investigating possibilities of innovation by the process
simplification and by adopting high technology easily and
successfully.

12. Product development such that marketing is decisive.
Yield equilibrium specific rates of fermentation and growth
profiles.
Product concentration in finished broth, fermentation cycle or
fermentation rate and product recovery process.

13. III. OBJECTIVES OF DESIGN
AIM
PROCEDURE
PROJECTED DESIGN
AND
DEVELOPMENT1 . Product development
2. Possibilities of producing
new type of microbial group
in a known fermenter design
3. Process improvement
ORGANIZATION STRUCTURE

14. ISOLATION OF FERMENTATION PRODUCTS
The innumerable products synthesized using industrial fermentations can't
directly be consumed/used, rather they need to be harvested, purified,
packaged and tested before they could be put to their actual use. This step is
called as “DOWN STREAMING”
It is an important step in the production of pharmaceuticals like:
1. Antibiotics such as penicillin
2. Hormones such as insulin and human growth hormone
3. Antibodies such as infliximab and rituximab
4. Vaccines
5. Antibodies and enzymes used for diagnostic purposes
6. Industrial enzymes
7. Natural fragrance and flavor compounds

15. Downstream processing is generally considered to be a specialized
field in biochemical/chemical engineering. It encompasses
harvesting, purification and packaging of a specific industrial
product synthesized in marketable quantities.
Analytical bioseparation refers to the separation or purification of
biological products at different scales of operation, done at
analytical scale, the purpose of which is to maintain the desired
quality of the product.
Downstream processing involves four important steps that result
in the following progressive improvements in purity and
concentration of the desired product…

16. 1. Removal of insoluble:
This step involves the harvest of the product in the form of
solute in a particle-free supernatant.
An example of this is the separation of cells, cell debris or any
other particulate matter from the spent fermentation broth
containing an antibiotic.
This can be achieved by various methods like filtration,
centrifugation, sedimentation, flocculation, electro-
precipitation, precipitation, and gravity settling. Other
methods such as grinding, homogenization, or leaching, may
be required for recovering products from solid sources like
plant and animal tissues.

17. 2. Product isolation
This step involves the removal of those components whose
properties differ significantly from that of the desired product.
For maximum products, the presence of water in them is the
main impurity, and consequently, product isolation essentially
involves drying and concentrating the product.
This step involves processes such as solvent extraction,
adsorption, ultra filtration, and precipitation.

18. 3. Product purification:
This step is performed for separating those contaminants whose
physical and chemical properties closely resemble those of the
product.
Since this is rather difficult to carry out, the steps involved at this
stage may be expensive, and may require sensitive and
sophisticated apparatus.
This stage alone may use up a significant fraction of the entire
downstream processing expenditure.
Examples of some of the methods used in this step are affinity
chromatography, size exclusion chromatography, reversed phase
chromatography, crystallization and fractional precipitation, all of
which are very specific techniques.

19. This is the last step that involves final processing steps that
end up with a stable, functional, easily transportable and
convenient product.
This step involves processes like crystallization, desiccation,
lyophilization and spray drying.
Depending on the type of product and its application,
polishing might even include product sterilization and
removal/deactivation of trace contaminants that may
compromise product safety. It might virus removal too.
4. Product polishing:

20. Not every product essentially requires all the above
mentioned steps.
Some products may be recovered by a combination of
just two steps.
For example, using affinity chromatography, one can
isolate and purify a product in a single step.

21. FERMENTATION PROCESS FOR PENICILLINS:
Penicillin was the first important commercial product
produced by an aerobic submerged fermentation.
Earlier it was made from the fungus, Penicillium notatum.
Nowadays different species, Penicillium chrysogenum and
better extraction processes are used.
It requires lactose, other sugars and a source of nitrogen (yeast
extract) in the medium to grow well.
It is a secondary metabolite so it is only produced during the
stationary phase.
It requires a batch fermenter, and a fed batch process is usually
used to prolong the stationary period and hence increase
production.

22. Downstream processing is relatively easy since penicillin is
secreted into the medium so there is no need to break open
the fungal cells.
However, the product needs to be very pure since it is to be
used as a medical therapeutic drug so it is dissolved and
precipitated as a potassium salt to separate it from other
substances of the medium.

24. The resulting penicillin is called as Penicillin G. It can be
chemically and enzymatic ally modified to a variety of penicillins
with slightly different properties.
These semi synthetic penicilins include:
1. Penicillin V
2. Penicillin O
3. Ampicillin
4. Amoxicillin

25. FERMENTATION PROCESS FOR STREPTOMYCIN:
Streptomycin is a secondary metabolite produced by
Streptomyces griseus.
Change in environment condition and substrate availability
influences final product.
A soya bean based medium is used along with glucose as a carbon
source.

26. PHASES DURING FERMENTATION
PHASE 1: Rapid growth producing mycelial biomass. Little
production of Streptomycin is obtained.
PHASE 2: Additional production of mycelium. Streptomycin
accumulates in the medium.
Proteolytic activity of the microbe releases NH3 to the medium
from the soybean meal, causing a rise in pH.
PHASE 3: Process has completed. Finally the mycelium is
separated by filtration and antibiotic recovered.
The glucose and NH3 released are consumed during this phase.
The pH remains fairly constant-between 7.6 and 9.0.

28. FERMENTAION PROCESS FOR TETRACYCLINES
Recently it has been discovered that microorganisms of the genus
Streptomyces which produce chlortetracycline, will also produce
tetracycline particularly if the chloride ion concentration of the
fermentation medium is kept low.
This can be done by providing fermentation media from which chloride
ions are excluded, either by making up the fermentation medium with
chloride-free components or by treating the medium with agents which re
move or sequester the chloride ions, thus making them unavailable for
the formation of chlortetracycline.
Unfortunately some of the most effective components of fermentation
media for the production of the tetracycline antibiotics contain
substantial quantities of chloride ions. Corn steep liquor is one of the
most effective nutrient substances for the production of the tetracycline
antibiotics as well as many other antibiotics. Apparently, this natural
material contains something that is especially desired by the fermenting
microorganism. Highest yields of antibiotic are, therefore, obtained when
a portion of corn steep liquor is included in the aqueous nutrient medium.

30. FERMENTATION PROCESS FOR VITAMIN B12
Large scale industrial production of vitamin B12 occurs via microbial
fermentation, predominantly utilizing Pseudomonas denitrificans,
Propionibacterium shermanii, or Sinorhizobium meliloti.
However, these strains have several shortcomings, such as long fermentation
cycles, complex and expensive media requirements, and a lack of suitable
genetic systems for strain engineering.

31. Carbon Sources:
Corn Steep Glucose, Beet Molasses, Soya bean Meal/Glucose
Nitrogen sources:
Ammonium phosphate , Ammonium hydroxide

32. Recently, engineers have shifted their attention to Escherichia coli
as a platform for vitamin B12 production.

33. THANK YOU
RAVLEEN KAUR
V SEMESTER
B.PHARMACY
UIPS,PU.