Penicillium chrysogenum is a fungus that produces penicillin, which is used to create several important beta-lactam antibiotics. It grows in filamentous hyphae containing spores called conidia. Industrially, P. chrysogenum is cultured in a fermentation process involving controlled temperature, pH, aeration and precursors to produce penicillin. The penicillin is then extracted from the fermentation broth using counter-current solvent extraction and purified through crystallization and other treatments. P. chrysogenum remains a primary industrial source of several types of medically important penicillin antibiotics.

1. Ubaid Tariq Bhat
M.Pharm Biotechnology
Delhi Pharmaceutical Science Research University
E. Mail: bhatubaid55@gmail.com
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3.  Also known as Penicillium notatum.
 It is common in temperate and subtropical regions and
can be found on salted food products, but it is mostly
found in indoor environments, especially in damp or
water-damaged buildings.
 It is the source of several β-lactam antibiotics, most
significantly penicillin which inhibits the biosynthesis
of bacterial cell walls affecting lysis of the cell.
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4.  Kingdom: Fungi
 Division: Ascomycota
 Class: Eurotiomycetes
 Order: Eurotiales
 Family: Trichocomaceae
 Genus: Penicillium
 Species: Chrysogenum
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5.  Penicillium chrysogenum exhibits typical eukaryotic
cell structure; it has a tubulin cytoskeleton which is
used for motility.
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TAM structure of P.
Chrysogenum
Structure of P. Chrysogenum

6.  This image displays the typical filamentous hyphae that
contain many conidia.
 The oblong structures in the image are conidia, the asexual
spores of the fungus.
 In P. chrysogenum, the conidia are blue to blue-green.
 These conidia are the cause of pathogenicity in humans as
in the cases of allergy and endophthalmitis.
 The conidia originate from complexes known as
conidiophores.
 The growth of conidiophores begins when a stalk sprouts
out of a foot cell.
 The stalk swells at the end and forms a vesicle. Sterigmata
form from the vesicle which give way to long chains of
conidia.
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7.  It produces the hydrophobic β-lactam compound
penicillin.
 Penicillium chrysogenum remains the primary producer
of Penicilian G and Penicilian V
 P. chrysogenum has been used industrially to produce
Penicilian G and Penicilian V and Xanthocillin X, and
to produce the enzymes polyamine oxidase,
phosphogluconate dehydrogenase, and glucose oxidase.
 Penicillium chrysogenum can be used to assist crops to
fight off other pathogenic species.
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8.  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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9. 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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10.  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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11. 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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12.  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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13. 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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14.  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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15.  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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