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“PENECILLIN PRODUCTION USING INDUSTRIAL STRAIN
Penicillium Chrysogenum SPECIES”
Presented by – Sayantan Ray (17/FT/015)
Introduction
We all have, at one point or the other, heard about penicillin. From it’s accidental discovery
and massive use in World War Ⅱ to the Nobel Prize for medicine in 1945 by it’s rightful
owners, penicillin has made it’s proof as the “Miracle Drug” that revolutionized the course
of the medical industry. Without further ado, lets’ take a look at an important aspect that
made penicillin accessible at large in it’s industrial production.
What is Penicillin?
Penicillin is a group of antibiotics, derived originally from common moulds known as penicillium
moulds; which includes penicillin G, penicillin V, Procaine Penicillin and Benzathine Penicillin.
Metabolism: Liver
Elimination half-life: 0.5 to 56 hours
Excretion: Kidney
First true naturally occurring antibiotic ever discovered: a great medical breakthrough.
Group of antibiotics produced by the Penicillium fungi. It is a group of closely related
compounds, not a single compound.
Example: Amoxicillin, Ampicillin, Phenoxymethylpenicillin
Around 50 drugs that are penicillin.
History of Penicillin
 1928: Scottish biologist, Alexander Fleming discovered that the Staphylococcus culture he had
mistakenly left growing in open was contaminated with a mould which had destroyed the
bacteria.
After isolating a sample and testing it, he found that it belongs to the Penicillium family.
Later the mould was classified as Penicillium notanum.
 1939: using Fleming’s work, two medical researchers, Howard Florey and Ernst Chain managed
to purify penicillin in a powered form.
 1943: They produced penicillin on a large scale. This helped immensely to treat casualties during
the WW Ⅱ that had bacterial infections due to their wounds. Ernst Chain
Alexander Fleming Penicillin Howard Florey Ernst Chain
How does Penicillin works?
 Inhibits the synthesis of peptidoglycan in cell walls.
β-Lactum of penicillin binds to the enzyme
transpeptidase, that is used in the formation of
peptidoglycan cross linking.
The enzyme is inhibited, thus inability to form
cross linking.
Cell wall is weakened causing osmotic imbalance
in the cell death.
 As human cells don’t have cell walls, penicillin
does not affect them.
Specific Condition for Penicillin Production
 Most penicillin form filamentous broths. This means they can be difficult to
mix due to their high viscosity of the broth can hinder oxygen transfer.
 As penicillin is an aerobic organism; so a solution for the viscosity and the
filamentous growth of penicillium species could be bubble columns (air lift
reactors) which would distribute the oxygen equally and also to agitate the
medium.
 The optimum pH for penicillin growth is 6.5 and it should be maintained
using pH controller and acid base reservoir.
 Strain stability problems (mutation) – careful strain maintenance is
required.
 Biomass doubling is about 6 hours – Provisions must be made.
Industrial Production of
Penicillin
The industrial production of penicillin can be
classified into two processes:
 Upstream Processing: Upstream
Processing referred to processes before
input to the fermenter and encompasses
any technology that leads to the synthesis
of a product. It includes the exploration,
development and production.
 Downstream Processing:
Downstream Processing referred to
processes done to purify the output of the
fermenter until it reaches to the desired
product, such as extraction and
purification of a product from
fermentation.
Simplified Flow Diagram of Industrial
Penicillin Production
INDUSTRIAL PENICILLIN PRODUCTION
1) Medium:- i) Corn Steep Liquor
ii) Glucose
iii) Salts: salts such as MgSO4, K3PO4 and Sodium Nitrates.
2) Heat Sterilisation: Medium is sterilised at high heat (121°C) and high pressure (30 psi), usually
through a holding tube or sterilized together in the fermenter.
3) Fermentation: The fermentation condition for the penicillin mold usually requires temperatures
20-24 °C , pH is kept at 6.5 and pressure is kept at 1.02 atm. Fungal cells are able to handle rotation
speed of around 200rpm to mix the cultures evenly.
4) Seed Culture: The addition of penicillium chrysogenum spores into a liquid medium which is
constantly aerated and agitated and carbon and nitrogen are added sparingly alongside. Typical
parameters such as pH, Temperature, rpm and oxygen concentration are observed for 40 hours until
penicillin begins to be secreted by the fungus. After 7 days pH increases to 8.0 and
penicillin production ceases.
5) Removal of biomass: Bio-separation is done by a Rotary vacuum filter. After filtration phosphoric acid is
entered to decrease pH from 8.0 to 6.5, to prevent loss of activity of penicillin.
6) Addition of solvent: Organic solvents such as amyl acetate/butyl acetate are added to dissolve the penicillin
present in the filtrate.
7) Centrifugal Extraction: Usually a disk centrifuge is used to centrifuge and separate the solid waste from the
liquid compound which contains the penicillin. Then the supernatant will be transferred in the downstream
process to continue the extraction.
8) Extraction: A series of extraction is done and at last penicillum-sodium salt is obtained from the liquid
material by basket centrifuge, in which solids are easily removed.
9) Fluid Bed Drying: In fluid bed drying, hot gas is pumped to remove all the moisture and much dried and
powdered form of penicillin salt is obtained.
10) Storage: Penicillin (Penicillin G) is stored in containers and kept in a dried environment and then it can be
modified chemically and enzymatically to make a variety of penicillins with slightly different properties.
Example: Penicillin V, Penicillin O, Ampicillin, Amoxycllin
Disk Centrifuge
Fluid Bed Dryer
Conclusion
o Antibiotics are probably the most important group of compounds synthesized
by industrial microorganisms.
o The best known and probably the most medically important antibiotics are the
Lactam, Penicillin and Chephalosporins.
o Penicillin exhibits the properties of a typical secondary metabolites.
o Penicillin is commercially produced usually via a fed batch process carried out
aseptically in stirred tank fermenters.
Basics on Penicilin production

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Basics on Penicilin production

  • 1. “PENECILLIN PRODUCTION USING INDUSTRIAL STRAIN Penicillium Chrysogenum SPECIES” Presented by – Sayantan Ray (17/FT/015)
  • 2. Introduction We all have, at one point or the other, heard about penicillin. From it’s accidental discovery and massive use in World War Ⅱ to the Nobel Prize for medicine in 1945 by it’s rightful owners, penicillin has made it’s proof as the “Miracle Drug” that revolutionized the course of the medical industry. Without further ado, lets’ take a look at an important aspect that made penicillin accessible at large in it’s industrial production.
  • 3. What is Penicillin? Penicillin is a group of antibiotics, derived originally from common moulds known as penicillium moulds; which includes penicillin G, penicillin V, Procaine Penicillin and Benzathine Penicillin. Metabolism: Liver Elimination half-life: 0.5 to 56 hours Excretion: Kidney First true naturally occurring antibiotic ever discovered: a great medical breakthrough. Group of antibiotics produced by the Penicillium fungi. It is a group of closely related compounds, not a single compound. Example: Amoxicillin, Ampicillin, Phenoxymethylpenicillin Around 50 drugs that are penicillin.
  • 4. History of Penicillin  1928: Scottish biologist, Alexander Fleming discovered that the Staphylococcus culture he had mistakenly left growing in open was contaminated with a mould which had destroyed the bacteria. After isolating a sample and testing it, he found that it belongs to the Penicillium family. Later the mould was classified as Penicillium notanum.  1939: using Fleming’s work, two medical researchers, Howard Florey and Ernst Chain managed to purify penicillin in a powered form.  1943: They produced penicillin on a large scale. This helped immensely to treat casualties during the WW Ⅱ that had bacterial infections due to their wounds. Ernst Chain Alexander Fleming Penicillin Howard Florey Ernst Chain
  • 5. How does Penicillin works?  Inhibits the synthesis of peptidoglycan in cell walls. β-Lactum of penicillin binds to the enzyme transpeptidase, that is used in the formation of peptidoglycan cross linking. The enzyme is inhibited, thus inability to form cross linking. Cell wall is weakened causing osmotic imbalance in the cell death.  As human cells don’t have cell walls, penicillin does not affect them.
  • 6. Specific Condition for Penicillin Production  Most penicillin form filamentous broths. This means they can be difficult to mix due to their high viscosity of the broth can hinder oxygen transfer.  As penicillin is an aerobic organism; so a solution for the viscosity and the filamentous growth of penicillium species could be bubble columns (air lift reactors) which would distribute the oxygen equally and also to agitate the medium.  The optimum pH for penicillin growth is 6.5 and it should be maintained using pH controller and acid base reservoir.  Strain stability problems (mutation) – careful strain maintenance is required.  Biomass doubling is about 6 hours – Provisions must be made.
  • 7. Industrial Production of Penicillin The industrial production of penicillin can be classified into two processes:  Upstream Processing: Upstream Processing referred to processes before input to the fermenter and encompasses any technology that leads to the synthesis of a product. It includes the exploration, development and production.  Downstream Processing: Downstream Processing referred to processes done to purify the output of the fermenter until it reaches to the desired product, such as extraction and purification of a product from fermentation. Simplified Flow Diagram of Industrial Penicillin Production
  • 8. INDUSTRIAL PENICILLIN PRODUCTION 1) Medium:- i) Corn Steep Liquor ii) Glucose iii) Salts: salts such as MgSO4, K3PO4 and Sodium Nitrates. 2) Heat Sterilisation: Medium is sterilised at high heat (121°C) and high pressure (30 psi), usually through a holding tube or sterilized together in the fermenter. 3) Fermentation: The fermentation condition for the penicillin mold usually requires temperatures 20-24 °C , pH is kept at 6.5 and pressure is kept at 1.02 atm. Fungal cells are able to handle rotation speed of around 200rpm to mix the cultures evenly. 4) Seed Culture: The addition of penicillium chrysogenum spores into a liquid medium which is constantly aerated and agitated and carbon and nitrogen are added sparingly alongside. Typical parameters such as pH, Temperature, rpm and oxygen concentration are observed for 40 hours until penicillin begins to be secreted by the fungus. After 7 days pH increases to 8.0 and penicillin production ceases.
  • 9. 5) Removal of biomass: Bio-separation is done by a Rotary vacuum filter. After filtration phosphoric acid is entered to decrease pH from 8.0 to 6.5, to prevent loss of activity of penicillin. 6) Addition of solvent: Organic solvents such as amyl acetate/butyl acetate are added to dissolve the penicillin present in the filtrate. 7) Centrifugal Extraction: Usually a disk centrifuge is used to centrifuge and separate the solid waste from the liquid compound which contains the penicillin. Then the supernatant will be transferred in the downstream process to continue the extraction. 8) Extraction: A series of extraction is done and at last penicillum-sodium salt is obtained from the liquid material by basket centrifuge, in which solids are easily removed. 9) Fluid Bed Drying: In fluid bed drying, hot gas is pumped to remove all the moisture and much dried and powdered form of penicillin salt is obtained. 10) Storage: Penicillin (Penicillin G) is stored in containers and kept in a dried environment and then it can be modified chemically and enzymatically to make a variety of penicillins with slightly different properties. Example: Penicillin V, Penicillin O, Ampicillin, Amoxycllin Disk Centrifuge Fluid Bed Dryer
  • 10. Conclusion o Antibiotics are probably the most important group of compounds synthesized by industrial microorganisms. o The best known and probably the most medically important antibiotics are the Lactam, Penicillin and Chephalosporins. o Penicillin exhibits the properties of a typical secondary metabolites. o Penicillin is commercially produced usually via a fed batch process carried out aseptically in stirred tank fermenters.