BOT 708

ASSIGNMENT 2

 Submitted to :
 Dr Abida Akram
 Submitted by:
 Aleeza Nawab
 Registration number:
 19-ARID-1987
 Semester :
 3rd
 Degree :
 MSc Botany [Morning]

ASSIGNMENT 2
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ASSIGNMENT 2

OUTLINE ]

Discovery of penicillin by Alexander Flemings

Research on Penicillin at Oxford University

Florey Experiment on Mice

Penicillin production during WWII

Increasing yield of penicillin

Role of pharmaceutical companies

[first and 2nd conference on penicillin production]

Scaling up production of penicillin ‘

Commercial production of penicillin

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ASSIGNMENT 2

How Penicillin was commercialized ?

.At Alexander Flemings laboratory [US department of agriculture centre for agriculture
utilization [November 19,1999.] Research was held at Peoria III . In that research there were 5
pharmaceutical companies took part in discovery of penicillin . research was done during world
war II , penicillin was introduced in 1940 and its therapeutic potential was known in UK because
of WWII [world war II]penicillin is very affective drug against serious infections caused by
streptococci and many others.. here we will cover following points

Discovery of penicillin by Amender Flemings.

Antibiotics.

Antibiotics are compounds produced by bacteria which are able of killing microbial species . this
is the reason behind the use of moldy bread to wounds by Egyptians but not till 1928. Before
penicillin was introduced there were problems of pneumonia and gonorrhea or rheumatoid
fever .[ infection caused by streptococci , which cause pain in joints]. It was the time during
world war II . all patients in hospitals were suffering from blood poisoning.[ it was due to
scratch or cut ] but no affective treatment [blood poisoning is the condition when bacteria
enters person’s blood stream and cause serious infection but no more than poison . it is now
called sepsis ]the first true antibiotic was discovered by Alexander Flemings .He was a Professor
of Bacteriology at Mary’s hospital in London. on September 3, 1928 Alexander Flemings . He
began to sort the Petri dishes containing colonies of staphylococci that cause sore throat and
many other infections . He noticed something unusual . Some colonies were dotted where
molds were growing and the zone around it don’t contains such type of bacteria . He found
that the juice can kill harmful bacteria . He asked his assistants to collect that juice . At that
times, they were only able to make crude extract . Alexander Flemings research was published
in June , 1929 by the name of BRITISH JOURNAL OF EXPERIMENTAL PATGHOLOGY . its ,main
aim was to collect penicillin insensitive bacteria from penicillin sensitive bacteria from mixed
culture .. so interest goes in penicillin extraction .so. London school of Hygiene and tropical
medicine . they also tried but failed.

Penicillin research at Oxford university .

Some worker turned penicillin into life saving drug. Howard Florey ,Ernst Chain and their
colleagues at[ Sir William Dumps School of Pathology]. The work started in 1939 there was a
need to process at 500litres per week of mold filtrate .they started tO grow in baths ,tanks and
food tins.. at that time they designed fermentation vessel to renew the broth .. for looking the
process after fermentation , team of girls was employed at dollar 2 . so oxford lab turned into
penicillin factory but the method they were using was not so efficient. Large amount of
penicillin was made in oxford lab. Norman Heatley+ Edward Abraham they both extracted
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penicillin from filtrate extracting it in amyl acetate and back to water by using countercurrent
system.

Filtrate _______<H20____-amyl acetate[counter current system]

Edward Abraham used newly discovered techniques and help to speed up the process by
Alumina color chromatography. he removed impurities in clinical trials.

Florey experiment on Mice

in 1940 Florey , he found that penicillin can protect the mice against deadly streptococci. On
Feb 12, 1941 Albert Alexander he was an old policeman , he was first recipient of oxford
penicillin. He got serious infection while pruning roses and was injected with penicillin that
show some recovery but when the supply of penicillin was limited he was dead after some
weeks.

Penicillin production in WWII

The chemical industry was fully absorbed during world war II . there was a need for large
amount of penicillin production. Florey with his colleagues travelled to US during summer
in 1941 ,Rockefeller foundation gave them support to know about the interest of pharmaceutical
companies on penicillin production..John Fulton put British colleagues to Robert Thorn a former
mycologist . he had authority on penicillin NRRL in Peoria in fermentation division.NRRl make
large amount of penicillin production possible .

Increasing yield of penicillin.

The director of NRRL was Orville he took responsibility to increase the yield of penicillin under
observation of Robert Coghill.he was chief of fermentation division in NRRL agreement was
done that Heatley would remain in Peoria he will share his experience with American colleagues
after some weeks , Andrew Moyer found that he could increases the yield by substituting
lactose with sucrose that was used by Oxford university in their culture medium. Moyer had
another finding that when we use corn steep that is the byproduct of corn wet milling process
and they found a way to use it and by this way they increased the penicillin production by
adding its precursors e=g phenyl acetic acid to fermentation medium . they realized that the
method used by oxford university is not so efficient so most ]efficient method would be us
growth in submerge culture so 2 staff members at NRRL Kenneth and Rapper .they screened
many penicillin strains and got more yield because this method was very successful . soil
samples were sent to NRRL from around the world . the most affective strain showed from
moldy cantaloupe strain was produced with help of X-rays at Carnegie institute when it was
exposed to UV rays at Wiscornsin university its productivity was increased still further,

Role of pharmaceutical companies in production of penicillin.

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Heatley still helping NRRL staff for starting the penicillin work .H.Florey visited many
pharmaceutical companies to know about interest in penicillin production but was very
disappointed about it. By his trip only 3 companies started the process of penicillin research .it
was before arrival of H florey .he meet his old friend Alfred Newton and VP of medical affairs
at university of Pennsylvania Richard was chair of committee on medical research CMR of
scientific research and development .Richard reached 4 drugs and found that Florey also had
interest in these and informed many pharmaceutical companies if they can serve nation for
increasing penicillin production’s

First conference

held by Richard in Washington on October 8 1941.the main aim of his meeting was to exchange
his information with government for progress of penicillin production. many workers
participated in this meeting included CMR representatives ,national research council ,US
department of agriculture including many others.

Second conference

It was held in New York in December 10 days after Pearl Harbor and US entry into 2nd world
war. The heads of meeting were Merck, Squibb, Pfizer.etc and many others. Robert Coghill
reported on success at NRRL with corn steep liquor was encouraging to industry leaders
present.it was decided that companies would do their research independently and will inform
about developments. By this way other companies also take interest Merk said that plant can
produce several hundred liters of penicillin per week usi9ng broth flask and tray . in December
Heatley joined the Merk research where he introduced Oxford cup plate method of penicillin
assay which then became standard industry world wide . from March 1942 penicillin was
produced under OSRD Auspices and treated the first patient. Other 10 cases were treated in June
1942and penicillin was supplied by Merk.

Scaling up penicillin production.

Many pharmaceutical companies played major role for scaling up production of penicillin to
manufacture level by using submerge fermentation. While doing all these Smith noticed the
difficulty of scaling up production because penicillin needs air to grow . aerating in a deep tank
can show many problems .or if sterile air is introduced it can cause more bubbling . Smith
introduced glycerol Monorichiolate as anti foaming agent .. so , it also needs new cooling system
and mixing technology to mash it. Lilly was successful but the penicillin produced 2/3rd could be
lost during purification. it was just because of heat sensitivity . so it was realized that extraction
must be don’t at low temperature . freeze drying under low temperature gave best results in
purifying penicillin to solid state .so penicillin was affective against infection caused by
streptococci and many others.

Commercial production of penicillin.

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In 1943 world war board took responsibility for penicillin production.. they selected 21
companies for that purpose .under direction of Albert Elder .,Merk Pfizer and Squibb they all
were major contributors . the penicillin then start to reach public , ut priority was given to
military use .production was increased in 1944. It jumped form 21 billion unit in 1943 to 1,663
billions unit in 1944 to more than 6.8 trillions in 1945 . manufacturing techniques were changed
and it start from 1 liter flask with less than 1% yield to 10,000 gallons tank with 80-90% yield
March 15 1945 it was distributed through usual channels ,1n 1944 annual production of
penicillin in US was 133,229billion units and price dropped form 20 dollar /100,000 unit in
1943 –less than 10 units .. many British companies moved over to deep tank fermentation to
meet needs. It was first reach public as prescription on June 1946.Abraham also worked for this
by using X-ray crystallography. its unique feature was established in 1945. It is 4 member highly
labile beta lactam ring .. in same year Alexander Flemings Howard Florey and Ernst chain were
awarded noble prize for penicillin research .many American chemist also helped for production
of penicillin .