project b pharma 6th sem. Penicillium mold naturally produces the antibiotic penicillin. ... Scientists learned to grow Penicillium mold in deep fermentation tanks by adding a kind of sugar and other ingredients. This process increased the growth of Penicillium.

2. School Of Studies in Pharmaceutical
Science
Jiwaji University Gwalior,
(474011)
Assignment On
-
Production of
Penicillin
SUBMITTED TO -
Mrs. Sangeeta Thakur
Ma'am
SUBMITTED BY -
Piyush Lodhi
B-Pharma (6th.Sem.)

3. CONTENTS
• Introduction
• Properties of penicillin
• Types of penicillin
• Basic structure of penicillin
• Commercial production of penicillin
• Downstream Processing
• Antibiotics Production Methods
• Conclusion
• References

4. Introduction :
• 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

5. • 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.

6. • 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, ect..
• Aqueous solution of penicillin are unstable and must be stored
under refrigeration.
• Penicillin is most stable in the pᴴ range of 6.0 to 6.5 and
reasonably stable over the pᴴ range of 5.5 to 7.5 .
Properties of penicillin :

7. Types of penicillin :
• Penicillin are compound of the general formula C₁₆H₁₈N₂O₅S –R, in
which R represents the radical or group that is different for each
day. The structural formula of the most common type ( F, G, X and k
) are given.
• Penicillin F, G, X and K are produced by strain of the penicillin
notatum – chrysogenum group of molds ; flavicidin ( flavicin) by
Aspergilus flavus ; and dihydro F penicillin (gigantic acid ) by
Aspergilus gigantic.
Basic structure of penicillin :
• The basic structure of the penicillin's is 6- aminopenicillenic acid
(6- APA) , composed of a thiozolidine ring fused with a β- lactam
ring whose 6- amino position carries a variety of acyl substituent's.

8. • The Inoculum : Master stock (spores) is the source of inoculum.
Spores (P. chrysogenum) from working stocks cultured are suspended
in water or non toxic lauryl sulphonate, then added to the flask
containing wheat bran and nutrient soln. A shake flask culture of 4
day old is inoculated into a seed tank for 3 days.
• The medium : Jackson in 1958 prepared a media for penicillin
production . The major constituent of typical medium includes,
Commercial production of penicillin :
1) Fermentable carbohydrate
1) Potassium dihydrogen phosphate
2) Organic nitrogen source
3) Phenyl acetic acid precursor
4) Edible oil
5) Calcium carbonate (act as buffer)
6) Pᴴ after sterilization
Corn steep liquor(3.5%),
lactose(3.5%),glucose(1%).
0.4%
0.25%
1%
5.5% to 6.0%

9. • Aeration (oxygen supply) : supply of oxygen in a bioreactor is
the limiting factor in penicillin biosynthesis. Aeration speed is
between 3.0 to 1.5 .
• Temperature : Temperature plays an important role in penicillin
production it should be mentioned at 28° C.
• Biomass production : production of penicillin depends upon
biomass production therefore, it is describle to have a high
biomass concentration in the vessel. It is achieved by increasing
the agitation rate and power.
• Pᴴ : It is maintained early neutrality by calcium and magnesium
carbonate in the medium by phosphate buffer. It is also controlled
by adding sodium hydroxide or sulphuric acid in the medium.

10. Production of penicillin
Learning objective:
• To be able to identify useful products from m
icroorganisms
• To be able to identify the microorganisms us
ed and the main stages in the production of p
enicillin.
• To be able to describe how Downstream proc
essing is carried out to extract and purify the
end-product of fermentation.

11. Downstream Processing
• Products in a fermenter are impure and dilute
, so need to be purified by downstream proce
ssing.
• This usually involves filtration to separate th
e microbial cells from the liquid medium, foll
owed by chemical purification and concentra
tion of the product
• Downstream processing can account for 50%
of the cost of a process.

12. • Antibiotics are antimicrobial agents pro
duced naturally by other microbes (usu
ally fungi or bacteria).
• The first antibiotic was discovered in 18
96 by Ernest Duchesne and "rediscover
ed" byAlexander Flemming in 1928 fro
m the filamentous fungus Penicilium no
tatum.
• The antibiotic substance, named penicil
lin, was not purified until the 1940s (by
Florey and Chain), just in time to be use
d at the end of the second world war.

13. Antibiotic production
• There are over 10 000 different antibiotics kn
own, but only about 200 in commercial
• use, since most new antibiotics are no better
than existing ones.
• There is a constant search for new antibiotic
s. Antibiotics are the most-prescribed drugs
and are big business.
• Finding a new antibiotic and getting it on to t
he market is a very long process and can tak
e 15 years.

14. Antibiotic Production Methods
• Antibiotics are produced on an industrial scale
using a variety of fungi and bacteria.
• Penicillin is produced by the fungus Penicillium
chrysogenum which requires lactose, other sug
ars, and a source of nitrogen (in this case a yea
st extract) in the medium to grow well.
• Like all antibiotics, penicillin is a secondary met
abolite, so is only produced in the stationary ph
ase.
• What sort of fermenter does it require?
• It requires a batch fermenter, and a fed batch pr
ocess is normally used to prolong the stationar
y period and so increase production.

15. • Downstream processing is relatively ea
sy since penicillin is secreted into the
medium (to kill other cells), so there is
no need to break open the fungal cells.
• However, the product needs to be very
pure, since it being used as a therapeut
ic medical drug, so it is dissolved and t
hen precipitated as a potassium salt to
separate it from other substances in th
e medium.

16. • Procedure :

17. Batch fermenter

18. • The resulting penicillin (called penicillin
G) can be chemically and enzymatically
modified to make a variety of penicillins
with slightly different properties.
• These semi-synthetic penicillins includ
e penicillin V, penicillin O, ampicillin an
d amoxycillin.

19. • Recovery and purification : Harvested culture broth
includes penicillin G along with a variety of other metabolites .
Vacuum filter is used for separation of mycelium from the broth on
a rotary .conversion of penicillin to the anionic form occurs at low
pᴴ (2.0 to 2.5).the lowering of pᴴ is done by adding phosphoric
acid or sulphuric acid. For removal of pigment and other
impurities from solvent containing penicillin it is treated with
active charcoal. The product is back extracted into water from
solvent by adding potassium or sodium hydroxide to form its sail.
The product of penicillin is then crystallized into sodium or
potassium penicillin.

20. • Antibiotics are probably the most important group of
compounds synthesized by industrial microorganisms.
• The best known and probably the most medically important
antibiotics are the β- lactam, penicillin and cephalosporin's.
• Penicillin exhibits the properties of a typical secondary
metabolites.
• Penicillin is commercially produced usually via a fed batch
process carried out aseptically in stirred tank fermenters.
Conclusion :

21. • John S Rockey, Michael J waites, Neil L Morgan and Gary
Higton, 2001, “Industrial Microbiology – an introduction”,
Black well publishing . Page no- 163 –168.
• Samuel C, Prescott and Cecil G Denn, 2011, “Industrial
Microbiology”, Agrobios(India). Page no- 721 -725.
• Tanuja Singh and S S Purohit, 2011, “Fermentation
Technology”, Agrobios(India). Page no – 195 – 205.
References :