In This PPt you will find every detail about Penicillin like history of penicillin Discovery of penicillin Manufacture of penicillin Advantage of penicillin Disadvantage of penicillin

1. RAKESH
B.Sc (Hons)Microbiology 3rd year
18081564027
Assignment
INDUSTRIAL MICROBIOLOGY
SUBMITTED TO:
DR.PARVINDER KAUR

2. Topics to be covered-
Introduction to Penicillin.
History and discovery of Penicillin.
Penicillin explanation.
Production Of Penicillin.
 Mode of action
 ADVERSE EFFECTS

3. INTODUCTION --
Antibiotics - Antibiotics are medicines that help stop infections caused by bacteria. They do
this by killing the bacteria or by keeping them from copying themselves or reproducing. The
word antibiotic means “against life.” Any drug that kills germs in your body is technically
an antibiotic.
“Antibiotic” this term was very first used by Selman Abraham Waksman in 1945.
Here are few examples of antibiotics---
 Penicillin - for example, phenoxymethylpenicillin, flucloxacillin and amoxicillin.
 Cephalosporin - for example, cefaclor, cefadroxil and cephalexin.
 Tetracycline - for example, tetracycline, doxycycline and lymecycline.

4. PENICILLIN
 Penicillin was the 1st antibiotic to be discovered by Alexander Fleming.
 It is still one of the best antibiotic available.
 It is an cell wall formation inhibitor in bacteria.
 It is produced by the Penicillium spp.

Its general structure Penicillin produced in 1943

5. Natural Penicillin Semi-synthetic Penicillin
Natural penicillin are produced by
microorganisms .
Partially by microbes and partially by
chemicals.
They can show allergic response. No allergic response.
They can only be injected in body. Can be taken as tablet form also,
Antimicrobial narrow spectrum. Antimicrobial broad spectrum.
Development of resistance by
microbes is more against these.
Development of resistance by
microbes is less against these.
No steric hindrance More steric hindrance.
Example- Penicillin F
Penicillin G
Penicillin X
Penicillin V
Penicillin K
Example-Ampicillin
Metacillin
Amoxycillin

7. ST.MARY HOSPITAL , LONDON
Lab Of ALEXANDER FLEMING
ORIGINAL
CULTURE PLATE SEEN
Curiosity
Discovery
1929
Noble prize sharers
1945
3 SEP
.1928

8. Dr. Alexander Fleming was returning to work from the holidays on 3 sep 1928when he started to sort
through petridishes containing Staphylococcus that had been left in the laboratory over the break. As he
was sorting, he found mold in one of the samples that was secreting a liquid. The liquid seemed to be
warding off the bacteria. Fleming and his colleges, revealed the new substance as a rare form of
Penicillium Notatum.
He discoverd penicillin in 1929.
The secretion was capable of killing a wide range of bacteria. Immediate work began on isolating a pure
form of the penicillin. Because the extracting process
was difficult, Fleming needed other scientist to carry out his work.
Two scientists from Oxford University, Earnest Chain and Howard Florey, were the men for the
job. They began using the pure form of Penicillin and converted it into a drug to be used in World
War II. Penicillin was used to rid infections of the wounded and saved many soldiers lives.
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."

9. Penicillin is an aerobic organism: so oxygen supply must be efficient in bio reactor.
The optimum pH for penicillium growth 6.5 maintain pH efficiently.
Biomass doubling is about 6h : provisions must be made to handle it.
Calcium carbonate or phosphates may be added as a
buffer. Sulfur compounds are sometimes added for additional yields
since penicillin contains sulfur.
The practice nowadays is to add the carbohydrate source
intermittently, i.e. using fed-batch fermentation.
Lactose is more slowly utilized and need not be added intermittently.
Glucose suppresses secondary metabolism and excess of it therefore
limits penicillin production.
The pH is maintained at between 6.8 and 7.4 by the automatic
addition of H2SO4 or NaOH as necessary.
Precursors of the appropriate side-chain are added to the fermentation.
Thus if benzyl penicillin is desired, phenylacetic acid is added. Phenyl
acetic acid is nowadays added continuously as too high an amount
inhibits the development of the fungus. High yielding strains of P.
chrysogenum resistant to the precursors have therefore been
developed.
Element Source
Carbohydrate/Car
bon
Glucose
Beet
Molasses/Lactose
Nitrogen corn steep liquor.
Cottonseed,
peanut,
linseed or
soybean meals

10. Early when the surface culture method was used, a variant of the original culture of Penicillium
notatum discovered by Sir Alexander Fleming was employed.
When however the production shifted to submerged cultivation, a strain of Penicillium
chrysogenum designated NRRL 1951 (after Northern Regional Research Laboratory of the
United States Department of Agriculture) discovered in 1943, was introduced.
In submerged culture it gave a penicillin yield of up to 250 Oxford Units (1 Oxford Unit =
0.5988 of sodium benzyl penicillin) which was two to three times more than given by
Penicillium notatum. A ‘super strain’ was produced from a variant of NRRL 1951 and
designated X 1612.
By ultraviolet irradiation of X-1612, a strain resulted and was named WISQ 176 after the
University of Wisconsin where much of the stain development work was done.
On further ultra violet irradiation of WISQ 176, BL3-D10 was produced, which produced only
75% as much penicillin as WISQ 176, but whose product lacked the yellow pigment the removal
of which had been difficult.
Present-day penicillin producing P. chrysogenum strains are far more highly productive than
their parents. They were produced through natural selection, and mutation using ultra violet
irradiation, x-irradiation or nitrogen mustard treatment. It was soon recognized that
there were several naturally occurring penicillins, viz. Penicillins G, X, F, and K
Strain of organism used in Penicillin Fermentation

11. In ancient times Penicillin was first produced commercially in stationary MAT culture.
But this kind of culture have a lot problems—
1.oxygen related problems(oxygen penetrates only
poorly the mycelial mat so this area remain devoid
to oxygen.
2.Large labour amount required
.

12. Why Penicillin G is Selected most.
Penicillin G (benzyl penicillin) was selected because—
1)It was markedly more effective against pyogenic cocci.
2)Higher yields were achieved by supplementing the medium with phenylacetic acid, analogues
(phenylalanine and phenethylaninie) of which are present in corn steep liquor used to grow penicillin
in the United States.

13. Penicillin fermentation can be divided into three phases
.The first phase (trophophase)during which rapid growth occurs, lasts forabout 30 hours during
which mycelia are produced.
The second phase (idiophase) lasts for five to seven days; growth is reduced
and penicillin is produced.
In the third phase, carbon and nitrogen sources are depleted,
antibiotic production ceases, the mycelia lyse releasing ammonia and the pH rises.
We use highly mutated strain of Penicillium chrysogenum
Avoid repeated subculturing.
They can be: agar slant/sterile soil/lyophilized stock/sporesuspension in liq.Nitorgen .
1)First we take stock culture with spores.
2)Then we will transfer this stock culture to conical flask.
3)Then transfer it to seed fermenter.
4)Then we transfer it to production fermenter.

14. Then in production fermenter keep it for (3-5 days) at 25 degree celcius.
Production fermenter is made up of steel of nichrome(Ni+Co)
Aeriation is must needed fermeter must have the impellers,agitators,spargers etc..
Penicillin G ---requires aseptic,submerged/fed-batch fermentation.
Medium requires — Carbon source - lactose sugar /molasses.
Nitrogen source - Corn steep liquor.
Vegetable oil –bot to produce much foam.
PHENYL ACETIC ACID AS PRECURSOR
Trace elements-salts,K,Na
pH-6.8-7.4
Absolute sterility is must.
Growth curve of bacteria and fungi is same.
Down stream process
Down stream is the process in which the product is extracted out and separated from the
waste .
FILTERATION- Rotatory vacuum filters are used to remove biomass.
The broth will be used now (culture filterate )

15. Solvent extraction-Penicillin gets dissolved in organic solvents amyl/ butylacetate
Then 2 stages of continuous counter current extraction is done at (0-3 degree
celcius) at 2.5-3 pH .Penicillin gets dissolved in acetate.
Re - extraction is done into an aqueous solution using phosphate buffer . We add a
source of sodium or potassium in buffer so that Penicillin-G gets cover into Na or
K salt.
Then we do centrifugation and wash respectively.
Then this is subjected to vacuum drying and we get 98-99% pure salt .
1mg of sodium salt of penicillin-G contains 1665 unit.
1 unit=0.6 micro gram.

18. Mechanisms of Action
All penicillin derivatives produce their bactericidal effects by inhibition of bacterial cell wall
synthesis.
Specifically, the cross linking of peptides on the polysaccharide chain is prevented.
If cell walls are improperly made cell walls allow water to flow into the cell causing it to burst.
Penicillin Bind (PBP) on the cell wall of susceptible bacteria Inhibits transpeptidation Prevents
peptidoglycan synthesis Cell wall deficient forms spheroplasts & filamentous forms Autolysis Cell death
(bactericidal action)
Binding to PBPs results in:
Inhibition of transpeptidase: transpeptidase catalyzes the cross-linking of the pentaglycine bridge with
the fourth residue (D-Ala) of the pentapeptide. The fifth residue (also D-Ala) is released during this
reaction. Spheroblasts are formed.
Structural irregularities: binding to PBPs may result in abnormal elongation, abnormal shape, cell wall
defects.

19. FLOW CHART
Penicillin
Bind (PBP)on cell wall of susceptible bacteria.
Inhibits transpeptidation
Prevents peptidoglycan synthesis
Cell wall deficient forms spheroploblasts and filamentous forms
Autolysis and cell death respectively

20. It is used in human therapeutic
It is used in animal feed.
It is used in labs to inhibit bacterial growth.
It is used in research also.

21. Pain at I.M injection site.
Nausea on oral ingestion.
Thromboplebitis (inflammatory process results as blood clot)
of Injected vein.

22. Q.1 Describe the production of Penicillin G. (6 Marks)
Q.2 Who coins the term “Antibiotic”. (1 Marks)
Q.3 Who discovered the Penicillin and write a brief note about it. (4 Marks)
Q.4 Which fungi was observed by the A.Fleming on his plate during penicillin discovery (2 Marks)

23. References
 Okafor N(2007)Modern Industrial Microbiology and Biotechnology, Ch-24 431-435p.g no 1st
edition.
 Casida (L.E) Industrial Microbiology Ch-17 223-247 page no.
 https://www.drugs.com/penicillin.html
 https://pharmaxchange.info/2011/04/mechanism-of-action-of-penicillin/
 https://www.drugs.com/penicillin.html
 https://www.slideshare.net/Vijaykumar1919/penicillins-
vk#:~:text=Mechanisms%20of%20Action%20%EF%81%AE%20All,the%20polysaccharide%20
chain%20is%20prevented.
 YOUTUBE VIDEOS LINKS
https://www.youtube.com/watch?v=7qeZLLhx5kU
 https://www.youtube.com/watch?v=a81nHSqQuvI
 https://www.youtube.com/watch?v=qBdYnRhdWcQ
 https://www.youtube.com/watch?v=-NZ3MSJbdi8

24. “Use us wisely otherwise
We will use you wisely”