history,production and Applications of Pencillin

1. Industrial Production of
Pencillin

2. Key Points and key terms
• Antibiotics are the secondary metabolites of microorganisms.
• During processing, the antibiotic must be extracted and purified to a crystalline
product.
• Useful antibiotics are often discovered using a screening process or a rational
design process.
• Antibiotic: Any substance that can destroy or inhibit the growth of bacteria and
similar microorganisms.
• Fermentation: Any of many anaerobic biochemical reactions in which an
enzyme (or several enzymes produced by a microorganism) catalyses the
conversion of one substance into another; especially the conversion (using
yeast) of sugars to alcohol or acetic acid with the evolution of carbon dioxide.
• Metabolite: Any substance produced by, or taking part in, a metabolic reaction.

4. History of Pencillin
 The first antibiotic was discovered in 1896 by Ernest Duchesne and
"rediscovered" by Alexander Flemming in 1928 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.
 The Nobel Prize in Physiology or Medicine 1945 was awarded jointly to Sir
Alexander Fleming, Ernst Boris Chain and Sir Howard Walter Florey ‘for the
discovery of penicillin and its curative effect in various infectious diseases’
 Penicillin was the first important commercial product produced by an aerobic,
submerged fermentation
 When penicillin was first made at the end of the second world war using the
fungus Penicilium notatum, the process made 1 mg dm‐3.
 Today, using a different species (P. chrysogenum) and a better extraction
procedures the yield is 50 g dm‐3.
 There is a constant search to improve the yield.

5.  Let’s see the reason
Why…..
Albert
Alexander
injury
became
infected
experimental
treatment
with partially
purified
penicillin
injected with
two hundred
milligrams
returned to
normal
treatment with
penicillin there was
not enough of the
wonder drug
scratched by the
thorn of a garden
rose.
Penicillin contributed in a major way to a
revolutionary change in medical treatment
which in turn changed the human lifestyle to
such an extent that diseases which were
common causes of death and disability before
penicillin are now rarely encountered.
This dramatic change to the every-day
experience of everyone on the planet is the
most remarkable aspect of the discovery and
introduction of penicillin.
More than any other scientific advance of modern
times, the penicillin story is able to stir interest and
imagination because this one fungal product simply
revolutionised medicine. It cannot cure everything
but penicillin can be used successfully to treat
pneumonia, gangrene, gonorrhoea, septicaemia
and osteomyelitis.
Diseases that were fatal and widespread, when
penicillin first came into use were relegated to
medical history. In the 1930s (and before) any injury
in which the skin was broken might become
infected with soil or air-borne bacteria and some of
these might grow beyond the level with which the
immune system could deal.

6. Properties of Pencillin
 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 .
 Optically active.
 Acid resistant.
 Hydrolyzed by hot inorganic acid .

7. Structure
 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.
 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.
Crystal structure of APS kinase

8. There are 6 types
•Penicillin G
•Penicillin V
•Penicillin F
•Penicillin X
•Penicillin K
•Penicillin O
Types of Penicillin
Penicillin F
Penicillin K
Penicillin X

9. : It encompasses any
technology that leads to the synthesis of a
product. Upstream includes the exploration,
development and production. The medium is
designed to provide the organism with all the
nutrients that it requires. Inoculation method-
submerged technique Spores -major source of
inoculum.
The extraction
and purification of a biotechnological product
from fermentation is referred to as downstream
processing.
Industrial Production of Penicillin
The industrial production of penicillin was broadly classified in to two processes namely,

10. Penicillin Production process:
Penicillin production is previously achieved by surface process ie. Solid state fermentation
and surface liquid ferementation. Now a days acommercial production is carried out by
fed batch process
Inoculum (Organism): Penicillium
chrysogenum (improved strain)
Inoculum preparation:
 For inoculum preparation, spore from
heavily sporulated working stocks are
suspended inwater or non-toxic wetting
agensts (sodium sulfonate 1: 10000)
 Theses spore are then added to flask
containing wheat bran and nutrient
solution for heavy sporulation
 Incubate for 5-7 days at 24C
 Spore are then transferred to seed tank
and incubated for 24-48 hours at 24C with
aeration and agitation for sufficient
mycelial growth
 These mycelia can be used for
production fermenter
Production fermentation:
 Method: fed-batch or batch
 Substrate: glucose, phenoxyacetic acid (fed component
used for production of side chain), Corn steep liquor,
Additional nitrogen source ie, soyameal, yeast extract,
Lactic acid, inorganic ions, growth factors
 Fermenter: stirred tank or air lift tank
 pH: set at 5.5 t0 6.0 which increased upto 7-7.5 (optimum)
due to liberation of NH3 gas and consumption of lactic
acid. If pH is 8 or more, CaCO3 or MgCO3 or phosphate
buffer is added
 temperature: 25-27 C
 aeration: 0.5-1 vvm (initially more, latter less O2 )
 agitation: 120-150 rpm)
 time: 3-5 days
 antiform: edible oil (0.25%)

11. Product recovery:
harvest broth from
fermenter tank by filtration
(rotary vaccum filtration)
chill to 5-10 C (because penicillin is highly reactive and destroyed by alkali and enzyme)
acidify filtrate to pH 2.0-2.5 with H2SO4 ( to convert penicillin to its anionic form)
extract penicillin from aqueous filtrate into butyl acetate or amyl acetate (at this very low pH as soon as
possible in centrifugal counter current extractor)
discard aqueous fraction
allow the organic solvent to pass through charcoal to remove impurities and extract
penicillin from butylacetate to 2% aqueous phosphate buffer at pH 7.5
acidify the aq. Fraction to pH 2-2.5 with mineral acid and re-extract
penicillin into fresh butylacetate ( it concentrated upto 80-100 times)
add potassium acetate to the solvent extract in a crystallization
tank to crystalize as potassium salt
recover crystal in filter centrifuge
sterilization
further processing
packaging

12. Process Fermentor

14. Trade names of penicillin in India
 PENTIDS -Nicholas Piramal India Ltd. (NPIL)
 FORTIFIED P.P -Nicholas Piramal India Ltd. (NPIL)
 BISTREPEN PAED-Alembic Ltd.
 BISTREPEN- Alembic Ltd.
 SODICILLIN Hindustan Antibiotics Ltd.
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)

15. Advantages Disadvantages
 Have excellent tissue penetration .
 Bactericidal against sensitive strains.
 Relatively nontoxic.
 Efficacious in the treatment of
infections.
 Inexpensive in comparision with other
antibiotics.
 Newer penicillin’s are resistant to
stomach acid , such as penicillin V or a
broader spectrum ,such as ampicillin
and amoxicillin.
 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.

16. Conclusion
 Worldwide sales of penicillin and other beta-lactam antibiotics is
now greater than $15 billion (U.S. dollars) per year. These sales
numbers exist despite the fact that cost is now at an all-time
low. Penicillin now costs $10 per kilogram versus $300 per kilogram
in 1953.
 Although Europe is the major producer of beta-lactam antibiotics,
newer manufacturing facilities are relocating to China and other
regions of Asia where labor and energy costs are lower.