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Production of Penicillin by Fermentation Technology

The document discusses the production of penicillin, an antibiotic discovered by Alexander Fleming in 1928, which targets microbial cell walls. It outlines the fermentation processes for penicillin production, emphasizing the transition from surface culture to submerged culture methods for efficiency. Key production conditions, media ingredients, and purification steps are also detailed to ensure optimal antibiotic yield and quality.

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10 views13 pages

Production of Penicillin by Fermentation Technology

The document discusses the production of penicillin, an antibiotic discovered by Alexander Fleming in 1928, which targets microbial cell walls. It outlines the fermentation processes for penicillin production, emphasizing the transition from surface culture to submerged culture methods for efficiency. Key production conditions, media ingredients, and purification steps are also detailed to ensure optimal antibiotic yield and quality.

Uploaded by

dhuravone
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We take content rights seriously. If you suspect this is your content, claim it here.
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Unit-V

Production of Penicillin
PHARMACEUTICA by Fermentation
L
Technology
BIOTECHNOLOGY
BP605T

Institute of Pharmaceutical Research, GLA University, Mathura, UP


• For antibiotics to function as a “magic bullet,” they usually must target
microbial structures or functions that are not shared with mammalian
structures or functions.

• The eukaryotic mammalian cell usually does not have a cell wall; instead, it
has only a plasma membrane.

• Even this membrane differs in composition from the plasma membrane of


prokaryotic cells.

• For this reason, the microbial cell wall is an attractive target for the action of
antibiotics.
Penicillin
• The term penicillin refers to a group of over 50 chemically related antibiotics.

• All penicillins have a common core structure containing a β-lactam ring called
the nucleus.

• Types are differentiated by the chemical side chains attached to their nuclei.

• Penicillins prevent the cross-linking of the peptidoglycans, which interferes


with the final stages of the construction of the cell walls, primarily of gram-
positive bacteria.

• Penicillins can be produced either naturally or semisynthetically.


• It was the first antibiotic which was discovered in 1896 by Ernest Duchesne
and "rediscovered" by Alexander Fleming in 1928.
• In 1928, a Scottish Biologist, Alexander Fleming is credited with the
discovery of penicillin.
• He discovered that the Staphylococcus culture which he had mistakenly left
growing in open was contaminated with a mould which had destroyed the
bacteria.
• After isolating a sample and by performing further tests he discovered that
it belonged to the Penicillium family, later the mould was classified as
Penicillium Notatum.
• In 1939, by using Fleming's work, two researchers, Howard Florey and
Ernest chain purified penicillin in a powdered form.
• In 1941, they successfully treated a human. Later in 1943, they produced
penicillin on a large scale which helped in treating persons during world
War ll that had bacterial infections due to their wounds.
Production of Penicillin
• The penicillin was originally manufactured by culturing strain of mould
Penicillium Chrysogenum because it is high yielding strain.
• So it is mostly used as production strain.
• Earlier the fermentation of Penicillium Chrysogenum is done by surface culture
process.
• In surface culture process the microorganisms were grown in the form of a pad
on the surface of a liquid medium in trays or flasks.
• But it has many disadvantages like
• Thousands of flasks were required to produce adequate amounts of antibiotic.
• There are many handling problems during processing like washing, sterilization,
filling, inoculation and bulking which made the process tedious and
economically non-viable.
• At present, submerged culturing is carried out to obtain the antibiotic.

• In submerged process the mould is allowed to grow deep in the liquid


medium in large fermenter which is constantly maintained under aeration
and agitation.

• So that the micro-organisms remain homogeneously suspended as single


cells, or in the form of very small aggregates or as colonies throughout the
liquid culture medium.
Penicillin may be produced by either
• Surface-culture method in which the mold is grown on the surface of shallow
layers of the fermentation medium.

• Submerged culture method in which the mold is grown submerged in the


fermentation medium in shake flasks, or deep tanks. Agitation and aeration of
the medium are essential in the submerged-culture method.

• Mold : Penicillium chrysogenum

• Master stock culture: The selected production strain of P. chrysogenum is


maintained in the form of a master.
Media:

Ingredients Concentration (g/lit.)

Cornsteep liquor (dry basis) 21.88

(NH4)2SO4 10.70

KH2PO4 2.74

Sodium phenyl acetate 15.90

Glucose 81.90

MgSO4.7H20, CaCO3, antifoam oil etc. q.s.


Conditions and production:
• Penicillin production is an aerobic process and therefore, a continuous supply
of oxygen to the growing culture is very essential.

• The required aeration rate is 0.5-1.0 vvm (vessel volume/min.). The


effectiveness of aeration can be enhanced by use of pressure, a tank pressure
of about 20 lb. per sq. in. may be satisfactory.

• The pH is maintained around 6.5.

• Optimal temperature is in the range of 25-27°C.

• Pencillin production is usually carried out by submerged processes.


Conditions and production………….
• Calcium carbonate is often used as buffering

• Multiple, flat-bladed disc turbine impellers, operated at a speed sufficient to


deliver 2-3 K.W. of power per kiloliter of aeration liquid, are generally
satisfactory.

• Antifoam agents such as tributyl citrate, octadecanol, and lard oil, prevent
excessive foam formation during the production of penicillin by submerged-
culture.

• Kept for 120-240 hr. for production


Isolation and Purification:
• The first step in the recovery process is the removal of mycelium or cells
by filtration or centrifuging.

• The second step is to remove the antibiotic from the spent production
medium by solvent extraction, adsorption or precipitation.

• Additional solvent extraction, distillation, sublimation, column


chromatography, or other methods accomplish purification.

• Finally the antibiotic is tested, as sayed and certified, as specified in the


relevant Pharmacopoeia.

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