2. INTRODUCTION
Penicillin is a secondary metabolite produced by certain bacteria,
which is used an antibiotic. .
‘Antibiotic’literally means ‘against life’–but antibiotics only kill life
that is harmful to living creatures ,i.e. bacteria.
A bacterial infection is caused by millions of tiny bacteria that
are trying to survive and in multiply the body . An antibiotic
attacks and kill these bacteria .
Before the development of penicillin , many people suffered and
died from bacterial infections that are no longer considered
dangerous today .
3. • Antibiotics are antimicrobial agents produced naturally by other
microbes (usually fungi or bacteria)
• The first antibiotic was discovered in 1896 by Ernest Duchesne and
in 1928 "rediscovered" by Alexander Fleming from the
filamentous fungus Penicilium notatum.
• The antibiotic substance, named penicillin, was not purified until
the 1940s (by Florey and Chain), just in time to be used at the end
of the second world war.
• Penicillin was the first important commercial product produced by
an aerobic, submerged fermentation.
• Penicillin is produced by the fungus Penicilium chrysogenum which
requires lactose, other sugars, and a source of nitrogen (in this case
a yeast extract) in the medium to grow well.
• Like all antibiotics, penicillin is a secondary metabolite, so is only
produced in the stationary phase
4. PROPERTIES OF PENICILLIN
.
It exhibits the properties of a typical secondary metabolites.
It active against certain Gram- positive bacteria in presence of
blood, pus and body fluids.
It is soluble in water. It is very soluble in acetone, ethyl alcohol
and ether and it is less soluble in benzene, chloroform, etc...
Aqueous solution of penicillin are unstable and must be stored
under refrigeration.
Penicillin ismost stable in the pᴴ range of 6.0 to 6.5 and
reasonably stable over the pᴴ range of 5.5 to 7.5 .
5. APPLICATION TO BIOTECHNOLOGY
It produces the hydrophobic β-lactam compound penicillin.
Penicillium chrysogenum remains the primary producer of
Penicillin G and Penicillin V
P. chrysogenum has been used industrially to produce
Penicillin G and Penicillin V and Xanthocillin X, and to
produce the enzymes polyamine oxidase, phospho gluconate
dehydrogenase, and glucose oxidase.
Penicillium chrysogenum can be used to assist crops to fight
off other pathogenic species
6. P.chrysogenum is high yielding strain and therefore most widely
used as production strain.
Inoculum Preparation:
Purpose is to develop a pure inoculum in an adequate amount.
To do so various sequential steps are necessary like:
1) A starter culture is needed for inoculation.
2) 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.
3) After about 24-28 hours, the content of the seed tanks is
transferred to the primary fermentation tanks.
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7. 4) All the bio parameters like temperature, pH, aeration,
agitation etc. should be properly maintained.
Bio parameters
pH: near 6.5
Temperature: 26°C to 28°C
Aeration: a continuous stream of sterilized air is
pumped into it.
Agitation: have baffles which allow constant agitation
(200rpm).
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8. Fermentation broth contains all the necessary elements
required for the proliferation of the microorganisms.
Generally, it contains a carbon source, nitrogen source,
mineral source, precrsors and antifoam agents.
Carbon Source
Lactose in a concentration of 6%.
Other carbohydrates like glucose & sucrose.
Complex as well as cheap sources like molasses, or
soya meal can also be used which are made up of
lactose and glucose sugars.
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9. Nitrogen Source
Ammonium salts such as ammonium sulphate,
ammonium acetate, ammonium lactate or ammonia gas
are used for this reason.
Sometime corn steep liquor may be used.
Mineral Source
These elements include phosphorus, sulphur,
magnesium, zinc, iron, and copper which generally
added in the form of water soluble salts.
Precursors
Various types of precursors are added into production
medium to produce specific type of penicillin.
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10. For example, if phenyl acetic acid is provided then only
penicillin-G will be produced but if hydroxy phenyl
acetic acid is provided then penicillin-X will be
produced.
Phenoxy acetic acid is provided as precursor for
penicillin-V production.
When corn steep liquor is provided as nitrogen source,
it also provides phenyl acetic acid derivatives; therefore
it is widely used in the production of penicillin-G.
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11. Anti-foam agents
Anti-foaming agents such as lard oil, octadecanol and
silicones are used to prevent foaming during
fermentation.
Recovery
The recovery of penicillin is carried out in three
successive stages:
1. Removal of mycelium
2. Counter current solvent extraction of penicillin
3. Treatment of crude extracts
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12. At harvest the fermentation broth is filtered on a rotatory
vacuum filter to remove the mycelium and other solids.
Phosphoric or sulfuric acids are added to lower the pH (2 to
2.5) in order to transform the penicillin to the anionic form.
Then the broth is directly extracted in a Podbielniak
Counter Current Solvent Extractor with an organic solvent
such as methyl isobutyl ketone, amyl acetate or butyl
acetate.
Penicillin is then again extracted into water from the
organic solvent by adding an adequate amount of potassium
or sodium hydroxide to form a salt of the penicillin.
The resulting aqueous solution is again acidified & re-
extracted with methyl isobutyl ketone.
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13. This shifts between water and solvent help in
purification of the penicillin.
The solvent extract is carefully back extracted with
NaOH and from this aqueous solution; various
procedures are utilized to cause the penicillin to
crystalize as sodium or potassium penicillinate.
The resulting crystalline penicillin salts are then
washed and dried.
Sometimes the crude extract of penicillin is passed out
from charcoal treatment to eliminate pyrogens; even
sterilization can also be done.
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16. rotating filter
filtrate fungal cells
Dissolve in butyl acetate animal feed
Potassium ions added to
Precipitate salt of penicillin
Wash, filter and dry
99.55% pure penicillin
19. ADVANTAGES
Have excellent tissue penetration.
Bactericidal against sensitivestrains.
Relatively nontoxic.
Efficacious in the treatment of infections.
Inexpensive in comparision with otherantibiotics.
Newer penicillin’s are resistant to stomach acid , such as penicillin V
or a broader spectrum ,such as ampicillin and amoxicillin.
DISADVANTAGES
Acid liability – most of these drugs are destroyed by gastric acid.
Lack of activity against most gram negative organisms.
Short duration of action.
Many patients experience GI upset.
Painful if given intramuscularly.
20. PENICILLIN MANUFACTURES &SUPPLIERS
Servo care life sciences pvt.ltd. (Chandigarh,
India)
Karan health care private limited (New Delhi,
India)
Alchemy medicine pvt.ltd. (Gurgaon,India)
Medicare remedies private limited (Navy Mumbai,
India)
