https://www.youtube.com/watch?v=aHXj7ONiEt8&t=25s Production of penicillin and citric acid

1. Pharma Knowledge
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2. What is anantibiotic?
The term antibiotic is refers to an organic compound that is
produced by one microorganism that, at great dilutions,
inhibits the growth of or kills another or even group of other
harmful microorganisms.
Antibiotics are available in various forms.
For example,
As ointment or cream to treat bacterial infection on the
surface.
As tablet/capsules/injection to treat internal infection.

3. Antibiotics are produced primarily by bacteria, Streptomycetes,
Nocardia and fungi. However, antibiotics produced by
Streptomycetes spp. Having greatest commercial applications.
On the basis of mode of action antibiotics can be divided as:
Antibiotics affecting cell wall
Antibiotics damaging cell membrane
Antibiotics interfering with protein synthesis
Antibiotics inhibiting nucleic acid synthesis
Antibiotics blocking cell metabolism
What is antibiotic

4. Penicillin
• The first discovered natural antibiotic was Penicillin which was
obtained from multicellular fungi, “Penicillium molds”.
• Penicillin is a group of compounds having 6-amino penicillinic
acid, contains ring like structure termed as a β-lactam ring.
• Penicillin are of two different types,
1. Natural Penicillin (directly harvested from the
penicillium mold).
2. Semi-synthetic Penicillin consists of the basic
Penicillin nucleus (6-APA), but with new side chain. It
includes Ampicillin, Methicillin, Penicillin V, etc.

5. Penicillin
• The first discovered natural antibiotic was Penicillin by Dr
Flemming; which was obtained from multicellular fungi,
“Penicillium molds”.
• Penicillin is a group of compounds having 6-amino penicillinic
acid, contains ring like structure termed as a β-lactam ring.
• Penicillin are of two different types,
1. Natural Penicillin (directly harvested from the
penicillium mold).
2. Semi-synthetic Penicillin consists of the basic
Penicillin nucleus (6-APA), but with new side chain. It
includes Ampicillin, Methicillin, Penicillin V, etc.

6. Production of Penicillin
Inoculum
• P. notatum & P. chrysogenum are two species used.
• P. chrysogenum is high yielding strain and therefore most widely
used as production strain.
• A starter culture transferred to an agar containing plate to allow
growth.
• After getting growth on solid media, one or two growth stages
should allowed in shaken flask cultures to create a suspension,
which can be transferred to seed tanks for further growth.
• Feed tank contain all the nutrients including Growth factors like
vitamins & amino acid as well as it may maintain temperature,
pH, agitation, aeration etc.

7. Media
Components Percent(%) Use
Lactose 3.5 to 6 C-sourse
Calcium Carbonate 1.0 buffer
Corn steep Liquor 3.5 N-source
Glucose 1.0 C-sourse)
Phenyl acetic acid 0.5 Precursor
Sodium hydrogen phosphate 0.4 catalyst
Edible oil 0.25
Bio parameters
• pH: near 6.5
• Temperature: 26°C to 28°C
• Aeration: a continuous stream of sterilized air is pumped into it.
• Agitation: achieved by turbine type impellers for constant agitation.
• Time period 5-6 days
Production of Penicillin

8. Recovery and purification
• Vacuum filter is used for separation of mycelium
from the broth on a rotary.
• The lowering of pᴴ is done by adding phosphoric
acid or sulphuric acid for conversion of penicillin to
the anionic form occurs at low pᴴ (2.0 to 2.5).
• For removal of pigment and other impurities from
solvent containing penicillin it is treated with active
charcoal.
• The product of penicillin is then crystallized into
sodium or potassium penicillin.
Production of Penicillin

9. Citric Acid
Citric acid is the most important commercial product,
which is found in almost all plant & animal tissues.
Citric acid is the most important organic acid produced in
tonnage and is extensively used in food and
pharmaceutical industries.
Citric acid is a weak organic acid found in citrus
fruits(lemon).
It is natural preservative and is also used to add an acidic
taste to food and soft drinks.
Million tonnes of citric acid are produced every year by
fermentation.

10. Inoculum
• The fungus Aspergillus niger is used.
• Mutant strain of Aspergillus niger is used for improved
production.
• It can tolerate high sugar concentration and low pH with
reduced synthesis of undesired byproducts.
Industrial fermentationprocess is carried out in three different ways as:
1. Surface fermentation
2. Submerged fermentation
3. Solid state fermentation
Production of Citric acid

11. Surface Fermentation
a. Solid Surface Fermentation
Carried out in some Asian countries like Japan using
A. niger.
Solid Substrate such as wheat bran or pulp from
sweet potato starch are used, as culture media.
Fermentation is done at pH is kept 4-5 at incubation
temp. of 280 C for 80-100 h.
Recovery and isolation is done using hot water .
Production of Citric acid

12. Surface Fermentation
b. Liquid Surface Fermentation
This is oldest method & carried out using liquid
nutrient such as beet carried out in aluminium trays
filled with nutrient media.
Inoculum in the form of spores sprayed.
Temperature spread 300 C
The pH of medium fall below 2 , as mycellium grows.
After 7-15 days , the mycellium is separated, pressed
mechanically and washed to get maximum amout of
citric acid.
Production of Citric acid

13. SUBMERGEDFERMENTATION:
• In this case, the strains are inoculated of about 15cm
depth in fermentation tank.
• The culture is enhanced by giving aeration using air
bubbles. And its allowed to grow for about 5 to 14
days at 27 to 33 degree Celsius.
• The citric acid produced in the fermentation tank and
it is purified.
Production of Citric acid

14. Solid statefermentation:
• It is simplest method for citric acid production.
• Solid state fermentation is also known as Koji
process, was first developed in Japan.
• Citric acid production reached a
maximum(88g/kg dry matter) when
fermentation as carried out with cassava
having initial moisture of 62% at 26degree
Celsius for 120 hours.
Production of Citric acid

15. Recovery
• The mycellium is separated by filtration of culture broth
and washing.
• Oxalic acid as unwanted byproduct is separated by
precipitation by adding lime.
• The liquid is transferred to recovery process
• Separation of citric acidis done at pH 7.2 and temp. 70-90
degree by precipitation.
• Further purification is done by dissolving citric acid in
sulfuric acid.
• Finally treated with activated charcoal, cation, anion
exchanger and crystallize
Production of Citric acid

16. Thank you all
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