Biotechnology in the general sense has been an important part of biology since the late 19th

century. With the industrialization of brewing and agriculture, chemists and biologists became
aware of the great potential of human-controlled biological processes. In particular, fermentation
proved a great boon to chemical industries. By the early 1970s, a wide range of biotechnologies
were being developed, from drugs like penicillin and steroids to foods like Chlorella and singlecell protein to gasoholas well as a wide range of hybrid high-yield crops and agricultural
technologies, the basis for the Green Revolution.[79]
The development of biochemistry and the recognition that most important biological processes
take place at the molecular level led to the rapid growth of the field of molecular biology, with
such fundamental results as the discovery of the structure of deoxyribonucleic acid (DNA), the
molecule carrying the genetic code. Modern medicine has profited from this explosion of
knowledge in biology and biochemistry, with new methods of treatment ranging from penicillin,
insulin, and a vast array of other drugs to pacemakers for weak hearts and implantation of
artificial or donated organs.
The use of antibiotics began with penicillin in 1928 and steroids were discovered in
1935.

Penicillin
Penicillin (PCN or pen) is a group of antibiotics which include penicillin G (intravenous use),
penicillin V (use by mouth), and procaine penicillin, and benzathine penicillin (intramuscular
use). Penicillin antibiotics were among the first medications to be effective against many
bacterial infections caused by staphylococci and streptococci. Penicillins are still widely used
today, though many types of bacteria have developed resistance following extensive use.
About 10% of people report that they are allergic to penicillin; however, up to 90% of this group
may not actually be allergic.[2] Serious allergies only occur in about 0.03%.[2] All penicillins are lactam antibiotics.
Penicillin was discovered in 1928 by Scottish scientist Alexander Fleming.[3] People began using
it to treat infections in 1942.[4] There are several enhanced penicillin families which are effective
against additional bacteria; these include the antistaphylococcal penicillins, aminopenicillins and
the antipseudomonal penicillins. They are derived from Penicillium fungi.[5]

Medical uses[edit]
The term "penicillin" is often used generically to refer to benzylpenicillin (penicillin G, the
original penicillin found in 1928), procaine benzylpenicillin (procaine penicillin), benzathine
benzylpenicillin (benzathine penicillin), and phenoxymethylpenicillin (penicillin V). Procaine
penicillin and benzathine penicillin have the same antibacterial activity as benzylpenicillin but
act for a longer period of time. Phenoxymethylpenicillin is less active against gram-negative

bacteria than benzylpenicillin.[6][7] Benzylpenicillin, procaine penicillin and benzathine penicillin

can be given by intravenous or intramuscular injections, but phenoxymethylpenicillin can be
given by mouth because of its acidic stability.[8]

Susceptibility[edit]
While the number of penicillin-resistant bacteria is increasing, penicillin can still be used to treat
a wide range of infections caused by certain susceptible bacteria, including Streptococci,
Staphylococci, Clostridium, and Listeria genera. The following list illustrates minimum
inhibitory concentration susceptibility data for a few medically significant bacteria:[9][10]

Listeria monocytogenes: from less than or equal to 0.06 g/ml to 0.25 g/ml

Neisseria meningitidis: from less than or equal to 0.03 g/ml to 0.5 g/ml

Staphylococcus aureus: from less than or equal to 0.015 g/ml to more than 32 g/ml

Adverse effects[edit]
Common adverse drug reactions ( 1% of people) associated with use of the penicillins include
diarrhoea, hypersensitivity, nausea, rash, neurotoxicity, urticaria, and superinfection (including
candidiasis). Infrequent adverse effects (0.11% of people) include fever, vomiting, erythema,
dermatitis, angioedema, seizures (especially in people with epilepsy), and pseudomembranous
colitis.[11]
About 10% of people report that they are allergic to penicillin; however, 90% of this group are
not actually allergic.[2] Serious allergies only occur in about 0.03%.[2]
Penicillin can also induce serum sickness or a serum sickness-like reaction in some individuals.
Serum sickness is a type III hypersensitivity reaction that occurs one to three weeks after
exposure to drugs including penicillin. It is not a true drug allergy, because allergies are type I
hypersensitivity reactions, but repeated exposure to the offending agent can result in an
anaphylactic reaction.[12]
Pain and inflammation at the injection site is also common for parenterally administered
benzathine benzylpenicillin, benzylpenicillin, and, to a lesser extent, procaine benzylpenicillin.
Although penicillin is still the most commonly reported allergy, less than 20% of people who
believe that they have a penicillin allergy are truly allergic to penicillin;[13] nevertheless, penicillin
is still the most common cause of severe allergic drug reactions. Significantly, there is an
immunologic reaction to Streptolysin S, a toxin released by certain killed bacteria and associated
with Penicillin injection, that can cause fatal cardiac syncope.[14]
Allergic reactions to any -lactam antibiotic may occur in up to 1% of patients receiving that
agent.[15] The allergic reaction is a Type I hypersensitivity reaction. Anaphylaxis will occur in

approximately 0.01% of patients.[11] It has previously been accepted that there was up to a 10%
cross-sensitivity between penicillin-derivatives, cephalosporins, and carbapenems, due to the
sharing of the -lactam ring.[16][17] Assessments in 2006 found no more risk for cross-allergy for
second-generation or later cephalosporins than the first generation. However, as a general risk,
research shows that all beta lactams have the intrinsic hazard of very serious hazardous reactions
in susceptible patients. Only the frequency of these reactions vary, based on the structure.[18][19]
Papers in 2006 showed that a major feature in determining frequency of immunological reactions
is the similarity of the side chains (e.g., first generation cephalosporins are similar to penicillins);
this is why the -lactams are associated with different frequencies of serious reactions (e.g.,
anaphylaxis).[20]

Mechanism of action

Bacteria that attempt to grow and divide in the presence of penicillin fail to do so, and instead
end up shedding their cell walls.

Penicillin and other -lactam antibiotics act by inhibiting penicillin-binding proteins, which
normally catalyze cross-linking of bacterial cell walls.
Bacteria constantly remodel their peptidoglycan cell walls, simultaneously building and breaking
down portions of the cell wall as they grow and divide. -Lactam antibiotics inhibit the
formation of peptidoglycan cross-links in the bacterial cell wall; this is achieved through binding

of the four-membered -lactam ring of penicillin to the enzyme DD-transpeptidase. As a

consequence, DD-transpeptidase cannot catalyze formation of these cross-links, and an
imbalance between cell wall production and degradation develops, causing the cell to rapidly die.
The enzymes that hydrolyze the peptidoglycan cross-links continue to function, even while those
that form such cross-links do not. This weakens the cell wall of the bacterium, and osmotic
pressure becomes increasingly uncompensatedeventually causing cell death (cytolysis). In
addition, the build-up of peptidoglycan precursors triggers the activation of bacterial cell wall
hydrolases and autolysins, which further digest the cell wall's peptidoglycans. The small size of
the penicillins increases their potency, by allowing them to penetrate the entire depth of the cell
wall. This is in contrast to the glycopeptide antibiotics vancomycin and teicoplanin, which are
both much larger than the penicillins.[21]
Gram-positive bacteria are called protoplasts when they lose their cell walls. Gram-negative
bacteria do not lose their cell walls completely and are called spheroplasts after treatment with
penicillin.
Penicillin shows a synergistic effect with aminoglycosides, since the inhibition of peptidoglycan
synthesis allows aminoglycosides to penetrate the bacterial cell wall more easily, allowing their
disruption of bacterial protein synthesis within the cell. This results in a lowered MBC for
susceptible organisms.[22]
Penicillins, like other -lactam antibiotics, block not only the division of bacteria, including
cyanobacteria, but also the division of cyanelles, the photosynthetic organelles of the
glaucophytes, and the division of chloroplasts of bryophytes. In contrast, they have no effect on
the plastids of the highly developed vascular plants. This supports the endosymbiotic theory of
the evolution of plastid division in land plants.[23]
The chemical structure of penicillin is triggered with a very precise, pH-dependent directed
mechanism, effected by a unique spatial assembly of molecular components, which can activate
by protonation. It can travel through bodily fluids, targeting and inactivating enzymes
responsible for cell-wall synthesis in gram-positive bacteria, meanwhile avoiding the
surrounding non-targets. Penicillin can protect itself from spontaneous hydrolysis in the body in
its anionic form, while storing its potential as a strong acylating agent, activated only upon
approach to the target transpeptidase enzyme and protonated in the active centre. This targeted
protonation neutralizes the carboxylic acid moiety, which is weakening of the -lactam ring N
C(=O) bond, resulting in a self-activation. Specific structural requirements are equated to
constructing the perfect mouse trap for catching targeted prey.[24]

Structure[edit]

Chemical structure of Penicillin G. The sulfur and nitrogen of the five-membered thiazolidine
ring are shown in yellow and blue respectively. The image shows that the thiazolidine ring and
fused four-membered -lactam are not in the same plane.
The term "penam" is used to describe the common core skeleton of a member of the penicillins.
This core has the molecular formula R-C9H11N2O4S, where R is the variable side chain that
differentiates the penicillins from one another. The penam core has a molecular weight of
243 g/mol, with larger penicillins having molecular weights near 450for example, cloxacillin
has a molecular weight of 436 g/mol. The key structural feature of the penicillins is the fourmembered -lactam ring; this structural moiety is essential for penicillin's antibacterial activity.
The -lactam ring is itself fused to a five-membered thiazolidine ring. The fusion of these two
rings causes the -lactam ring to be more reactive than monocyclic -lactams because the two
fused rings distort the -lactam amide bond and therefore remove the resonance stabilisation
normally found in these chemical bonds.[25]

History[edit]
Discovery

Alexander Fleming, who is credited with discovering penicillin in 1928.

Sample of penicillium mould presented by Alexander Fleming to Douglas Macleod, 1935
Starting in the late 19th century there had been many accounts by scientists and physicians on the
antibacterial properties of the different types of moulds including the mould penicillium but they
were unable to discern what process was causing the effect.[26] The effects of penicillium mould
would finally be isolated in 1928 by Scottish scientist Alexander Fleming, in work that seems to
have been independent of those earlier observations.[27] Fleming recounted that the date of his
discovery of penicillin was on the morning of Friday 28 September 1928.[28] The traditional
version of this story describes the discovery as a serendipitous accident: in his laboratory in the
basement of St Mary's Hospital in London (now part of Imperial College), Fleming noticed a
Petri dish containing Staphylococcus that had been mistakenly left open was contaminated by
blue-green mould from an open window, which formed a visible growth.[29] There was a halo of
inhibited bacterial growth around the mould. Fleming concluded that the mould released a
substance that repressed the growth and caused lysing of the bacteria.[30]
Once Fleming made his discovery he grew a pure culture and discovered it was a Penicillium
mould, now known to be Penicillium notatum. Fleming coined the term "penicillin" to describe

the filtrate of a broth culture of the Penicillium mould. Fleming asked C. J. La Touche to help
identify the mould, which he incorrectly identified as Penicillium rubrum (later corrected by
Charles Thom). He expressed initial optimism that penicillin would be a useful disinfectant,
because of its high potency and minimal toxicity in comparison to antiseptics of the day, and
noted its laboratory value in the isolation of Bacillus influenzae (now called Haemophilus
influenzae).[29][31]
Fleming was a famously poor communicator and orator, which meant his findings were not
initially given much attention.[29] He was unable to convince a true chemist to help him extract
and stabilize the antibacterial compound found in the broth filtrate. Despite the lack of a true
chemist, he remained interested in the potential use of penicillin and presented a paper entitled
"A Medium for the Isolation of Pfeiffer's Bacillus" to the Medical Research Club of London,
which was met with little interest and even less enthusiasm by his peers. Had Fleming been more
successful at making other scientists interested in his work, penicillin for medicinal use would
possibly have been developed years earlier.[29]
Despite the lack of interest of his fellow scientists, he did conduct several experiments on the
antibiotic substance he discovered. The most important result proved it was nontoxic in humans
by first performing toxicity tests in animals and then on humans. His following experiments on
penicillin's response to heat and pH allowed Fleming to increase the stability of the compound.[31]
The one test that modern scientists would find missing from his work was the test of penicillin
on an infected animal, the results of which would likely have sparked great interest in penicillin
and sped its development by almost a decade.[29]

Medical application[edit]

Florey (pictured), Fleming and Chain shared a Nobel Prize in 1945 for their work on penicillin.

In 1930, Cecil George Paine, a pathologist at the Royal Infirmary in Sheffield, attempted to use
penicillin to treat sycosis barbae, eruptions in beard follicles, but was unsuccessful. Moving on to
ophthalmia neonatorum, a gonococcal infection in infants, he achieved the first recorded cure
with penicillin, on November 25, 1930. He then cured four additional patients (one adult and
three infants) of eye infections, and failed to cure a fifth.[32][33][34]
In 1939, Australian scientist Howard Florey (later Baron Florey) and a team of researchers (Ernst
Boris Chain, Arthur Duncan Gardner, Norman Heatley, M. Jennings, J. Orr-Ewing and G.
Sanders) at the Sir William Dunn School of Pathology, University of Oxford made progress in
showing the in vivo bactericidal action of penicillin. In 1940, they showed that penicillin
effectively cured bacterial infection in mice.[35][36] In 1941, they treated a policeman, Albert
Alexander, with a severe face infection; his condition improved, but then supplies of penicillin
ran out and he died. Subsequently, several other patients were treated successfully.[37]

Mass production[edit]
A technician preparing penicillin in 1943
By late 1940, the Oxford team under Howard Florey had devised a method of mass-producing
the drug, but yields remained low.[37] In 1941, Florey and Heatley travelled to the US in order to
interest pharmaceutical companies in producing the drug and inform them about their process.[37]
Florey and Chain shared the 1945 Nobel Prize in Medicine with Fleming for their work.
The challenge of mass-producing this drug was daunting. On March 14, 1942, the first patient
was treated for streptococcal septicemia with US-made penicillin produced by Merck & Co.[38]
Half of the total supply produced at the time was used on that one patient. By June 1942, just
enough US penicillin was available to treat ten patients.[39] In July 1943, the War Production
Board drew up a plan for the mass distribution of penicillin stocks to Allied troops fighting in
Europe.[40] The results of fermentation research on corn steep liquor at the Northern Regional
Research Laboratory at Peoria, Illinois, allowed the United States to produce 2.3 million doses in
time for the invasion of Normandy in the spring of 1944. After a worldwide search in 1943, a
mouldy cantaloupe in a Peoria, Illinois market was found to contain the best strain of mould for
production using the corn steep liquor process.[41] In 19411944, Jasper H. Kane developed the
practical, deep-tank fermentation method for production of large quantities of pharmaceuticalgrade penicillin. Large-scale production resulted from the development of a deep-tank
fermentation plant by chemical engineer Margaret Hutchinson Rousseau.[42] As a direct result of
the war and the War Production Board, by June 1945, over 646 billion units per year were being
produced.[40]

Penicillin was being mass-produced in 1944.

G. Raymond Rettew made a significant contribution to the American war effort by his techniques
to produce commercial quantities of penicillin.[43] During World War II, penicillin made a major
difference in the number of deaths and amputations caused by infected wounds among Allied
forces, saving an estimated 12%15% of lives.[citation needed] Availability was severely limited,
however, by the difficulty of manufacturing large quantities of penicillin and by the rapid renal
clearance of the drug, necessitating frequent dosing. Methods for mass production of penicillin
were patented by Andrew Jackson Moyer in 1945.[44][45][46] Florey had not patented penicillin,
having been advised by Sir Henry Dale that doing so would be unethical.[37]
Penicillin is actively excreted, and about 80% of a penicillin dose is cleared from the body within
three to four hours of administration. Indeed, during the early penicillin era, the drug was so
scarce and so highly valued that it became common to collect the urine from patients being
treated, so that the penicillin in the urine could be isolated and reused.[47] This was not a
satisfactory solution, so researchers looked for a way to slow penicillin excretion. They hoped to
find a molecule that could compete with penicillin for the organic acid transporter responsible for
excretion, such that the transporter would preferentially excrete the competing molecule and the
penicillin would be retained. The uricosuric agent probenecid proved to be suitable. When
probenecid and penicillin are administered together, probenecid competitively inhibits the
excretion of penicillin, increasing penicillin's concentration and prolonging its activity.
Eventually, the advent of mass-production techniques and semi-synthetic penicillins resolved the
supply issues, so this use of probenecid declined.[47] Probenecid is still useful, however, for
certain infections requiring particularly high concentrations of penicillins.[11]
After World War II, Australia was the first country to make the drug available for civilian use. In
the U.S., penicillin was made available to the general public on March 15, 1945.[48]

Dorothy Hodgkin determined the chemical structure of penicillin.

Structure determination and total synthesis[edit]

Dorothy Hodgkin's model of penicillin's structure.

In 1945 the chemical structure of penicillin was determined using X-ray crystallography by
Dorothy Crowfoot Hodgkin, who was also working at Oxford.[49] She later received the Nobel
prize for this and other structure determinations.
Chemist John C. Sheehan at the Massachusetts Institute of Technology (MIT) completed the first
chemical synthesis of penicillin in 1957.[50][51][52] Sheehan had started his studies into penicillin
synthesis in 1948, and during these investigations developed new methods for the synthesis of
peptides, as well as new protecting groupsgroups that mask the reactivity of certain functional
groups.[52][53] Although the initial synthesis developed by Sheehan was not appropriate for mass
production of penicillins, one of the intermediate compounds in Sheehan's synthesis was 6aminopenicillanic acid (6-APA), the nucleus of penicillin.[52][54][55][page needed] Attaching different groups
to the 6-APA 'nucleus' of penicillin allowed the creation of new forms of penicillin.

Developments from penicillin[edit]

The narrow range of treatable diseases or "spectrum of activity" of the penicillins, along with the
poor activity of the orally active phenoxymethylpenicillin, led to the search for derivatives of
penicillin that could treat a wider range of infections. The isolation of 6-APA, the nucleus of
penicillin, allowed for the preparation of semisynthetic penicillins, with various improvements
over benzylpenicillin (bioavailability, spectrum, stability, tolerance).
The first major development was ampicillin in 1961. It offered a broader spectrum of activity
than either of the original penicillins. Further development yielded -lactamase-resistant
penicillins, including flucloxacillin, dicloxacillin, and methicillin. These were significant for
their activity against -lactamase-producing bacterial species, but were ineffective against the
methicillin-resistant Staphylococcus aureus (MRSA) strains that subsequently emerged.[citation needed]
Another development of the line of true penicillins was the antipseudomonal penicillins, such as
carbenicillin, ticarcillin, and piperacillin, useful for their activity against Gram-negative bacteria.
However, the usefulness of the -lactam ring was such that related antibiotics, including the
mecillinams, the carbapenems and, most important, the cephalosporins, still retain it at the center
of their structures.[56]

Production[edit]

A 1957 fermentor (bioreactor) used to grow Penicillium mould.

Penicillin is a secondary metabolite of certain species of Penicillium and is produced when
growth of the fungus is inhibited by stress. It is not produced during active growth. Production is
also limited by feedback in the synthesis pathway of penicillin.[citation needed]
-ketoglutarate + AcCoA homocitrate L--aminoadipic acid L-lysine + -lactam
The by-product, L-lysine, inhibits the production of homocitrate, so the presence of exogenous
lysine should be avoided in penicillin production.
The Penicillium cells are grown using a technique called fed-batch culture, in which the cells are
constantly subject to stress, which is required for induction of penicillin production. The
available carbon sources are also important: Glucose inhibits penicillin production, whereas
lactose does not. The pH and the levels of nitrogen, lysine, phosphate, and oxygen of the batches
must also be carefully controlled.[citation needed]

The biotechnological method of directed evolution has been applied to produce by mutation a
large number of Penicillium strains. These techniques include error-prone PCR, DNA shuffling,
ITCHY, and strand-overlap PCR.
Semisynthetic penicillins are prepared starting from the penicillin nucleus 6-APA.

Biosynthesis[edit]

Penicillin G biosynthesis
Overall, there are three main and important steps to the biosynthesis of penicillin G
(benzylpenicillin).

The first step is the condensation of three amino acidsL--aminoadipic acid, Lcysteine, L-valine into a tripeptide.[57][58][59] Before condensing into the tripeptide, the amino
acid L-valine must undergo epimerization to become D-valine.[60][61] The condensed
tripeptide is named -(L--aminoadipyl)-L-cysteine-D-valine (ACV). The condensation
reaction and epimerization are both catalyzed by the enzyme -(L--aminoadipyl)-Lcysteine-D-valine synthetase (ACVS), a nonribosomal peptide synthetase or NRPS.

The second step in the biosynthesis of penicillin G is the oxidative conversion of linear
ACV into the bicyclic intermediate isopenicillin N by isopenicillin N synthase (IPNS),
which is encoded by the gene pcbC.[57][58] Isopenicillin N is a very weak intermediate,
because it does not show strong antibiotic activity.[60]

The final step is a transamidation by isopenicillin N N-acyltransferase, in which the aminoadipyl side-chain of isopenicillin N is removed and exchanged for a phenylacetyl
side-chain. This reaction is encoded by the gene penDE, which is unique in the process of
obtaining penicillins.[57]

History of penicillin
From Wikipedia, the free encyclopedia

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Penicillin core structure, where "R" is the variable group.

The factual accuracy of part of this article is disputed. The dispute is about
Most of the examples cited in this table are not supported by evidence that 1) the
molds involved were penicillin-producing strains, or 2) that the mentioned
treatments were actually effective. See Talk. . Please help to ensure that disputed
statements are reliably sourced. See the relevant discussion on the talk page. (June
2014) (Learn how and when to remove this template message)

Alexander Fleming was the first to suggest the Penicillium mould must secrete an antibacterial
substance, and the first to concentrate the active substance which he named penicillin in 1928,
and during the next twelve years he grew and distributed the original mould, but he was not the
first to use its properties in medicine. Howard Florey organized a team which involved in the
mass production of penicillin include Ernst Chain, Howard Florey and Norman Heatley.
Year

Location

Descriptions
Many ancient cultures, including the ancient Egyptians, Greeks and in
Egypt,
ancient India, already used moulds and other plants to treat infection.[1]
Ancient
Greece & This worked because some moulds produce antibiotic substances.
times
India
However, they could not identify or isolate the active component in the
moulds.
"traditional" Russia
Russian peasants used warm soil as treatment for infected wounds.[2]
Soldiers in the army of king Dutugemunu (161137 BC) are recorded
to have stored oil cakes (a traditional Sri Lankan sweetmeat) for long
periods in their hearth lofts before embarking on their campaigns, in
c. 150 BC Sri Lanka
order to make a poultice of the cakes to treat wounds.[citation needed] It is
assumed that the oil cakes served the dual functions of desiccant and
antibacterial.[clarification needed]
Wet bread was mixed with spider webs (containing spores) to treat
1600s
Poland
wounds. The technique was mentioned by Henryk Sienkiewicz in his
1884 book With Fire and Sword.
The idea of using mould as a form of treatment was recorded by
1640
England
apothecaries, such as John Parkington, King's Herbarian, who
advocated the use of mould in his book on pharmacology.[citation needed]

1870

United
Kingdom

1871

United
Kingdom

1874

United
Kingdom

1875

United
Kingdom

1875
1877

France

1887

France

1895

Italy

1897

France

Sir John Scott Burdon-Sanderson, who started out at St. Mary's

Hospital 18521858 and as lecturer there 18541862 observed that
culture fluid covered with mould would produce no bacteria.
Burdon-Sanderson's discovery prompted Joseph Lister, an English
surgeon and the father of modern antisepsis to investigate that urine
samples contaminated with mould did not allow the growth of bacteria.
He also described the antibacterial action on human tissue of what he
called Penicillium glaucum.[3] A nurse at King's College Hospital whose
wounds did not respond to any antiseptic, was then given another
substance that cured her, and Lister's registrar informed her that it was
called Penicillium.
William Roberts observed that bacterial contamination is generally
absent in cultures of the mould Penicillium glaucum.
John Tyndall followed up on Burdon-Sanderson's work and
demonstrated to the Royal Society the antibacterial action of the
Penicillium fungus.[4]
Bacillus anthracis was shown to cause anthrax. This was the first
demonstration that a specific bacterium caused a specific disease.
Louis Pasteur and Jules Francois Joubert observed that cultures of the
anthrax bacilli, when contaminated with moulds, became inhibited.
Some references say that Pasteur identified the strain as Penicillium
notatum.[citation needed]
Garr found similar results.
Vincenzo Tiberio, physician of the University of Naples published a
research about a mould in a water well in Arzano, Italy that had an
antibacterial action.[5][6][7][8]
Ernest Duchesne at cole du Service de Sant Militaire in Lyon
independently discovered healing properties of a Penicillium glaucum
mould, even curing infected guinea pigs from typhoid. He published a
dissertation[9][10][11] in 1897 but this was ignored by the Institut Pasteur.
However Duchesne was himself using a discovery made by Arab stable
boys, who were using moulds to cure sores on horses. He did not claim
that the mould contained any antibacterial substance, only that the
mould somehow protected the animals.

Duchesne cured typhoid, but the penicillin isolated by Fleming

does not cure typhoid.

Duchesne injected a mould with the fungus Penicillium

glaucum. In contrast, Fleming isolated the substance penicillin
from the mould Penicillium notatum.

The term Penicillium glaucum was used as a catch-all phrase at

the time for different fungi, but not for Penicillium notatum and
the mould was unfortunately not preserved, which makes it

impossible to be certain today which fungus might have been

responsible for the cure, and consequently, even less certain
which substance was responsible.
Andre Gratia and Sara Dath observed a fungal contamination in one of
their Staphylococcus aureus cultures that was inhibiting the growth of
1920
Belgium the bacterium. They identified this as a species of Penicillium and
presented their observations as a paper. There was little attention to this
paper.
An Institut Pasteur scientist, Costa Rican Clodomiro Picado Twight
1923
Costa Rica
recorded the antibiotic effect of Penicillium.
Scottish biologist Sir Alexander Fleming noticed a halo of inhibition of
bacterial growth around a contaminant blue-green mould on a
Staphylococcus plate culture. He concluded that the mould was
releasing a substance that was inhibiting bacterial growth. He grew a
United
pure culture of the mould and discovered that it was Penicillium
1928
Kingdom notatum. With help from a chemist, he concentrated what he later
named "penicillin". During the next twelve years, he grew and
distributed the original mould, unsuccessfully trying to get help from
any chemist that had enough skill to make a stable form of it, for mass
production.
Cecil George Paine, a pathologist at the Royal Infirmary in Sheffield,
attempted to treat sycosis (eruptions in beard follicles) with penicillin
but was unsuccessful, probably because the drug did not penetrate deep
United
1930
enough. Moving on to ophthalmia neonatorum, a gonococcal infection
Kingdom
in babies, he achieved the first cure on 25 November 1930. He cured
four patients (one adult, the others babies) of eye infections, although a
fifth patient was not so lucky.[12]
In Oxford, Howard Walter Florey organized his large and very skilled
United
biochemical research team, notable among them Ernst Boris Chain and
1938
Kingdom Norman Heatley, to undertake innovative work to produce a stable
penicillin.
Peoria, Illinois: Moyer, Coghill and Raper at the USDA Northern
Regional Research Laboratory (NRRL) developed methods for
19411943 USA
industrialized penicillin production and isolated higher-yielding strains
of the Penicillium fungus.[13][14]
Brooklyn, New York: Jasper H. Kane and other Pfizer scientists
19411944 USA
developed the practical, deep-tank fermentation method for production
of large quantities of pharmaceutical-grade penicillin.[15]
United
Oxford: Using X-ray crystallographic analysis, Dorothy Hodgkin
1945
Kingdom elucidated the correct chemical structure of penicillin.[16]
Kundl, Tyrol: Hans Margreiter and Ernst Brandl of Biochemie (now
1952
Austria
Sandoz) developed the first acid-stable penicillin for oral
administration, Penicillin V.[17]

1957

USA

Chemist John C. Sheehan at the Massachusetts Institute of Technology

(MIT) completed the first chemical synthesis of penicillin in 1957.[18]

Discovery and Development of Penicillin

International Historic Chemical Landmark
Designated November 19, 1999, at the Alexander Fleming Laboratory Museum in London, U.K.
Also recognized at the U.S. Department of Agriculture National Center for Agricultural
Utilization Research in Peoria, Ill., and the five American pharmaceutical companies that
contributed to penicillin production research during WWII: Abbott Laboratories, Lederle
Laboratories (now Pfizer, Inc.), Merck & Co., Inc., Chas. Pfizer & Co. Inc. (now Pfizer, Inc.) and
E.R. Squibb & Sons (now Bristol-Myers Squibb Company).
Commemorative Booklet (PDF)
The introduction of penicillin in the 1940s, which began the era of antibiotics, has been
recognized as one of the greatest advances in therapeutic medicine. The discovery of penicillin
and the initial recognition of its therapeutic potential occurred in the United Kingdom, but, due
to World War II, the United States played the major role in developing large-scale production of
the drug, thus making a life-saving substance in limited supply into a widely available medicine.

Alexander Flemings Discovery of Penicillin

Penicillin heralded the dawn of the antibiotic age. Before its introduction there was no effective
treatment for infections such as pneumonia, gonorrhea or rheumatic fever. Hospitals were full of
people with blood poisoning contracted from a cut or a scratch, and doctors could do little for
them but wait and hope.
Antibiotics are compounds produced by bacteria and fungi which are capable of killing, or
inhibiting, competing microbial species. This phenomenon has long been known; it may explain
why the ancient Egyptians had the practice of applying a poultice of moldy bread to infected
wounds. But it was not until 1928 that penicillin, the first true antibiotic, was discovered by
Alexander Fleming, Professor of Bacteriology at St. Mary's Hospital in London.
Returning from holiday on September 3, 1928, Fleming began to sort through petri dishes
containing colonies of Staphylococcus, bacteria that cause boils, sore throats and abscesses. He
noticed something unusual on one dish. It was dotted with colonies, save for one area where a
blob of mold was growing. The zone immediately around the moldlater identified as a rare
strain of Penicillium notatumwas clear, as if the mold had secreted something that inhibited
bacterial growth.
Fleming found that his "mold juice" was capable of killing a wide range of harmful bacteria,
such as streptococcus, meningococcus and the diphtheria bacillus. He then set his assistants,

Stuart Craddock and Frederick Ridley, the difficult task of isolating pure penicillin from the mold
juice. It proved to be very unstable, and they were only able to prepare solutions of crude
material to work with. Fleming published his findings in the British Journal of Experimental
Pathology in June 1929, with only a passing reference to penicillin's potential therapeutic
benefits. At this stage it looked as if its main application would be in isolating penicillininsensitive bacteria from penicillin-sensitive bacteria in a mixed culture. This at least was of
practical benefit to bacteriologists, and kept interest in penicillin going. Others, including Harold
Raistrick, Professor of Biochemistry at the London School of Hygiene and Tropical Medicine,
tried to purify penicillin but failed.

Penicillin Research at Oxford University

It was Howard Florey, Ernst Chain and their colleagues at the Sir William Dunn School of
Pathology at Oxford University who turned penicillin from a laboratory curiosity into a lifesaving drug. Their work on the purification and chemistry of penicillin began in earnest in 1939,
just when wartime conditions were beginning to make research especially difficult. To carry out
a program of animal experiments and clinical trials the team needed to process up to 500 liters a
week of mold filtrate. They began growing it in a strange array of culture vessels such as baths,
bedpans, milk churns and food tins. Later, a customized fermentation vessel was designed for
ease of removing and, to save space, renewing the broth beneath the surface of the mold. A team
of "penicillin girls" was employed, at 2 a week, to inoculate and generally look after the
fermentation. In effect, the Oxford laboratory was being turned into a penicillin factory.
Meanwhile, biochemist Norman Heatley extracted penicillin from huge volumes of filtrate
coming off the production line by extracting it into amyl acetate and then back into water, using a
countercurrent system. Edward Abraham, another biochemist who was employed to help step up
production, then used the newly discovered technique of alumina column chromatography to
remove impurities from the penicillin prior to clinical trials.
In 1940, Florey carried out vital experiments, showing that penicillin could protect mice against
infection from deadly Streptococci. Then, on February 12, 1941, a 43-year old policeman, Albert
Alexander, became the first recipient of the Oxford penicillin. He had scratched the side of his
mouth while pruning roses, and had developed a life-threatening infection with huge abscesses
affecting his eyes, face, and lungs. Penicillin was injected and within days he made a remarkable
recovery. But supplies of the drug ran out and he died a few days later. Better results followed
with other patients though and soon there were plans to make penicillin available for British
troops on the battlefield.
War-time conditions made industrial production of penicillin difficult. A number of British
companies, including Glaxo (now GlaxoSmithKline) and Kemball Bishop, a London firm later
bought by Pfizer, took up the challenge.

Penicillin Production in the United States during WWII

Substantial amounts of penicillin would be needed for the extensive clinical trials required to
confirm the promise of the early results and to provide adequate supplies of the drug for
therapeutic use if it did live up to its potential. Florey recognized that large-scale production of
penicillin was probably out of the question in Britain, where the chemical industry was fully
absorbed in the war effort. With the support of the Rockefeller Foundation, Florey and his
colleague Norman Heatley traveled to the United States in the summer of 1941 to see if they
could interest the American pharmaceutical industry in the effort to produce penicillin on a large
scale.
Yale physiologist John Fulton helped to put his British colleagues in touch with individuals who
might be able to assist them in their goal. They were referred to Robert Thom of the Department
of Agriculture, a foremost mycologist and authority on the Penicillium mold, and eventually to
the Department's Northern Regional Research Laboratory (NRRL) in Peoria, Illinois, because of
the expertise of its Fermentation Division. This contact proved to be crucial to the success of the
project, as the NRRL was a key contributor of innovations that made large-scale production of
penicillin possible.

Increasing the Yield of Penicillin

Orville May, Director of the NRRL, agreed to have the Laboratory undertake a vigorous program
to increase penicillin yields under the direction of Robert Coghill, Chief of the Fermentation
Division. It was agreed that Heatley would remain in Peoria to share his expertise with his
American colleagues. Within a few weeks, Andrew Moyer found that he could significantly
increase the yield of penicillin by substituting lactose for the sucrose used by the Oxford team in
their culture medium. Shortly thereafter, Moyer made the even more important discovery that the
addition of corn-steep liquor to the fermentation medium produced a ten-fold increase in yield.
Corn-steep liquor was a by-product of the corn wetmilling process, and the NRRL, in an attempt
to find a use for it, tried it in essentially all of its fermentation work. Later, the Peoria laboratory
increased the yield of penicillin still further by the addition of penicillin precursors, such as
phenylacetic acid, to the fermentation medium.
It was recognized that the Oxford group's method of growing the mold on the surface of a
nutrient medium was inefficient, and that growth in submerged culture would be a superior
process. In submerged culture fermentation, the mold is grown in large tanks in a constantly
agitated and aerated mixture, rather than just on the surface of the medium. Florey's Penicillium
culture, however, produced only traces of penicillin when grown in submerged culture. Under the
direction of Kenneth Raper, staff at the NRRL screened various Penicillium strains and found
one that produced acceptable yields of penicillin in submerged culture.
Soon a global search was underway for better penicillin producing strains, with soil samples
being sent to the NRRL from around the world. Ironically, the most productive strain came from
a moldy cantaloupe from a Peoria fruit market. A more productive mutant of the so-called

cantaloupe strain was produced with the use of X-rays at the Carnegie Institution. When this
strain was exposed to ultraviolet radiation at the University of Wisconsin, its productivity was
increased still further.

U.S. Pharmaceutical Companies Support Production

While Norman Heatley remained in Peoria helping the NRRL staff to get the penicillin work
started, Howard Florey visited various pharmaceutical companies to try to interest them in the
drug. Although Florey was disappointed in the immediate results of his trip, three of the
companies (Merck, Squibb and Lilly) had actually conducted some penicillin research before
Florey's arrival and Pfizer seemed on the verge of investigating the drug as well. At this time,
however, the promise of penicillin was still based on only limited clinical trials.
Florey next visited his old friend Alfred Newton Richards, then vice president for medical affairs
at the University of Pennsylvania. More importantly, Richards was chair of the Committee on
Medical Research (CMR) of the Office of Scientific Research and Development (OSRD). The
OSRD had been created in June, 1941, to assure that adequate attention was given to research on
scientific and medical problems relating to national defense. Richards had great respect for
Florey and trusted his judgment about the potential value of penicillin. He approached the four
drug firms that Florey indicated had shown some interest in the drug (Merck, Squibb, Lilly and
Pfizer) and informed them that they would be serving the national interest if they undertook
penicillin production and that there might be support from the federal government.
Richards convened a meeting in Washington, D.C., on October 8, 1941, to exchange information
on company and government research and to plan a collaborative research program to expedite
penicillin production. In addition to representatives of the CMR, the National Research Council
and the U.S. Department of Agriculture, participants included research directors Randolph T.
Major of Merck; George A. Harrop of the Squibb Institute for Medical Research; Jasper Kane of
Pfizer; and Y. SubbaRow of Lederle. The next CMR penicillin conference, held in New York in
December, ten days after Pearl Harbor and U.S. entry into the Second World War, was more
decisive. At this meeting, which was attended by the heads of Merck, Squibb, Pfizer and Lederle,
as well as the company research directors, Robert Coghill's report on the success at the NRRL
with corn steep liquor was encouraging to the industry leaders present.
As Coghill later recalled, George W. Merck, who had been pessimistic about the possibility of
producing adequate quantities of penicillin given the constraints of available fermentation
techniques and yields,"...immediately spoke up, saying that if these results could be confirmed in
their laboratories, it was possible to produce the kilo of material for Florey, and industry would
do it!". It was agreed that although the companies would pursue their research activities
independently, they would keep the CMR informed of developments, and the Committee could
make the information more widely available (with the permission of the company involved) if
that were deemed in the public interest.

Although there was some concern that investments in fermentation processes might be wasted if
a commercially-viable synthesis of penicillin were developed, other companies also began to
show an interest in the drug. Some firms worked out collaborative agreements of their own (e.g.,
Merck and Squibb in February 1942, joined by Pfizer in September). Merck's pilot plant
continued to produce several hundred liters of penicillin culture per week using both flasks and
tray, and in December, Heatley joined the Merck research staff for several months, where he
introduced the Oxford cup plate method of penicillin assay, which soon became a standard
method industry-wide. By March 1942 enough penicillin had been produced under OSRD
auspices to treat the first patient (Mrs. Ann Miller, in New Haven, Connecticut); a further ten
cases were treated by June 1942, all with penicillin supplied by Merck & Co., Inc.

"The discovery and development of penicillin 1928-1945 commemorative booklet produced by

the National Historic Chemical Landmarks program of the American Chemical Society in 1999
(PDF).

Scaling-up Penicillin Production

Pharmaceutical and chemical companies played an especially important role in solving the
problems inherent in scaling up submerged fermentation from a pilot plant to a manufacturing
scale. As the scale of production increased, the scientists at Merck, Pfizer, Squibb and other
companies faced new engineering challenges. Pfizer's John L. Smith captured the complexity and
uncertainty facing these companies during the scale-up process: "The mold is as temperamental
as an opera singer, the yields are low, the isolation is difficult, the extraction is murder, the
purification invites disaster, and the assay is unsatisfactory."

Because penicillin needs air to grow, aerating the fermentation mixture in deep tanks presented a
problem. When corn steep liquor was used as the culture medium, bubbling sterile air through
the mixture caused severe foaming. Squibb solved this problem by introducing glyceryl
monoricinolate as an anti-foaming agent. Submerged fermentation also required the design of
new cooling systems for the vats and new mixing technology to stir the penicillin mash
efficiently.
Lilly was particularly successful in making the mold synthesize new types of penicillin by
feeding precursors of different structure. Once the fermentation was complete, recovery was also
difficult; as much as two-thirds of the penicillin present could be lost during purification because
of its instability and heat sensitivity. Extraction was done at low temperatures. Methods of
freeze-drying under vacuum eventually gave the best results in purifying the penicillin to a
stable, sterile, and usable final form.
The steps of fermentation, recovery and purification and packaging quickly yielded to the
cooperative efforts of the chemical scientists and engineers working on pilot production of
penicillin. On March 1, 1944, Pfizer opened the first commercial plant for large-scale production
of penicillin by submerged culture in Brooklyn, New York.
Meanwhile, clinical studies in the military and civilian sectors were confirming the therapeutic
promise of penicillin. The drug was shown to be effective in the treatment of a wide variety of
infections, including streptococcal, staphylococcal and gonococcal infections. The United States
Army established the value of penicillin in the treatment of surgical and wound infections.
Clinical studies also demonstrated its effectiveness against syphilis, and by 1944, it was the
primary treatment for this disease in the armed forces of Britain and the United States.

The real story behind penicillin

In a monthly column for PBS NewsHour, Dr. Howard Markel revisits moments that
changed the course of modern medicine on their anniversaries, like the development of
penicillin on Sept. 28, 1928. Above: Jean-Claude Fide is treated with penicillin by his
mother in 1948. Photo by Bert Hardy/Picture Post
The discovery of penicillin, one of the worlds first antibiotics, marks a true turning point
in human history when doctors finally had a tool that could completely cure their
patients of deadly infectious diseases.
Many school children can recite the basics. Penicillin was discovered in London in
September of 1928. As the story goes, Dr. Alexander Fleming, the bacteriologist on duty
at St. Marys Hospital, returned from a summer vacation in Scotland to find a messy lab
bench and a good deal more.
Upon examining some colonies of Staphylococcus aureus, Dr. Fleming noted that a
mold called Penicillium notatum had contaminated his Petri dishes. After carefully
placing the dishes under his microscope, he was amazed to find that the mold
prevented the normal growth of the staphylococci.

Sir Alexander Fleming (1881 1955), studying a test tube

culture with a hand lens. Photo by Chris Ware/Getty Images.

It took Fleming a few more weeks to grow enough of the persnickety mold so that he
was able to confirm his findings. His conclusions turned out to be phenomenal: there
was some factor in the Penicillium mold that not only inhibited the growth of the bacteria
but, more important, might be harnessed to combat infectious diseases.
As Dr. Fleming famously wrote about that red-letter date: When I woke up just after
dawn on September 28, 1928, I certainly didnt plan to revolutionize all medicine by
discovering the worlds first antibiotic, or bacteria killer. But I guess that was exactly
what I did.
Fourteen years later, in March 1942, Anne Miller became the first civilian patient to be
successfully treated with penicillin, lying near death at New Haven Hospital in
Connecticut, after miscarrying and developing an infection that led to blood poisoning.
But there is much more to this historic sequence of events.
Actually, Fleming had neither the laboratory resources at St. Marys nor the chemistry
background to take the next giant steps of isolating the active ingredient of the
penicillium mold juice, purifying it, figuring out which germs it was effective against, and
how to use it. That task fell to Dr. Howard Florey, a professor of pathology who was
director of the Sir William Dunn School of Pathology at Oxford University. He was a
master at extracting research grants from tight-fisted bureaucrats and an absolute
wizard at administering a large laboratory filled with talented but quirky scientists.
This landmark work began in 1938 when Florey, who had long been interested in the
ways that bacteria and mold naturally kill each other, came across Flemings paper on
the penicillium mold while leafing through some back issues of The British Journal of
Experimental Pathology. Soon after, Florey and his colleagues assembled in his wellstocked laboratory. They decided to unravel the science beneath what Fleming called
penicilliums antibacterial action.

A petri-dish of penicillin showing its inhibitory effect on some

bacteria but not on others. Photo by Keystone Features/Getty Images.

One of Floreys brightest employees was a biochemist, Dr. Ernst Chain, a Jewish
German migr. Chain was an abrupt, abrasive and acutely sensitive man who fought
constantly with Florey over who deserved credit for developing penicillin. Despite their
battles, they produced a series of crude penicillium-mold culture fluid extracts.
During the summer of 1940, their experiments centered on a group of 50 mice that they
had infected with deadly streptococcus. Half the mice died miserable deaths from
overwhelming sepsis. The others, which received penicillin injections, survived.
It was at that point that Florey realized that he had enough promising information to test
the drug on people. But the problem remained: how to produce enough pure penicillin to
treat people. In spite of efforts to increase the yield from the mold cultures, it took 2,000
liters of mold culture fluid to obtain enough pure penicillin to treat a single case of sepsis
in a person.
In September 1940, an Oxford police constable, Albert Alexander, 48, provided the first
test case. Alexander nicked his face working in his rose garden. The scratch, infected
with streptococci and staphylococci, spread to his eyes and scalp. Although Alexander
was admitted to the Radcliffe Infirmary and treated with doses of sulfa drugs, the
infection worsened and resulted in smoldering abscesses in the eye, lungs and
shoulder. Florey and Chain heard about the horrible case at high table one evening and,
immediately, asked the Radcliffe physicians if they could try their purified penicillin.
After five days of injections, Alexander began to recover. But Chain and Florey did not
have enough pure penicillin to eradicate the infection, and Alexander ultimately died.

A laboratory technician examining flasks of penicillin culture,

taken by James Jarche for Illustrated magazine in 1943.

Another vital figure in the lab was a biochemist, Dr. Norman Heatley, who used every
available container, bottle and bedpan to grow vats of the penicillin mold, suction off the
fluid and develop ways to purify the antibiotic. The makeshift mold factory he put
together was about as far removed as one could get from the enormous fermentation
tanks and sophisticated chemical engineering that characterize modern antibiotic
production today.
In the summer of 1941, shortly before the United States entered World War II, Florey
and Heatley flew to the United States, where they worked with American scientists in
Peoria, Ill., to develop a means of mass producing what became known as the wonder
drug.
Aware that the fungus Penicillium notatum would never yield enough penicillin to treat
people reliably, Florey and Heatley searched for a more productive species.
One hot summer day, a laboratory assistant, Mary Hunt, arrived with a cantaloupe that
she had picked up at the market and that was covered with a pretty, golden mold.
Serendipitously, the mold turned out to be the fungus Penicillium chrysogeum, and it
yielded 200 times the amount of penicillin as the species that Fleming had described.
Yet even that species required enhancing with mutation-causing X-rays and filtration,
ultimately producing 1,000 times as much penicillin as the first batches from Penicillium
notatum.
In the war, penicillin proved its mettle. Throughout history, the major killer in wars had
been infection rather than battle injuries. In World War I, the death rate from bacterial
pneumonia was 18 percent; in World War II, it fell, to less than 1 percent.

This is the penicillin table in a U.S. evacuation hospital in

Luxembourg in 1945. Photo by Photo12/UIG.

From January to May in 1942, 400 million units of pure penicillin were manufactured. By
the end of the war, American pharmaceutical companies were producing 650 billion
units a month.
Ironically, Fleming did little work on penicillin after his initial observations in 1928.
Beginning in 1941, after news reporters began to cover the early trials of the antibiotic
on people, the unprepossessing and gentle Fleming was lionized as the discoverer of
penicillin. And much to the quiet consternation of Florey, the Oxford groups
contributions were virtually ignored.
That problem was partially corrected in 1945, when Fleming, Florey, and Chain but
not Heatley were awarded the Nobel Prize in Physiology or Medicine. In his
acceptance speech, Fleming presciently warned that the overuse of penicillin might lead
to bacterial resistance.
In 1990, Oxford made up for the Nobel committees oversight by awarding Heatley the
first honorary doctorate of medicine in its 800-year history.
Maybe this September 28, as we celebrate Alexander Flemings great accomplishment,
we will recall that penicillin also required the midwifery of Florey, Chain and Heatley, as
well as an army of laboratory workers.

Penicillin is one of the earliest discovered and widely used antibiotic agents, derived from the
Penicillium mold. Antibiotics are natural substances that are released by bacteria and fungi into
the their environment, as a means of inhibiting other organisms - it is chemical warfare on a
microscopic scale.

Sir Alexander Fleming

Born August 6, 1881 in Darvel, Scotland

Died March 11, 1955 in London, England

In 1928, Sir Alexander Fleming observed that colonies of the bacterium Staphylococcus aureus
could be destroyed by the mold Penicillium notatum, proving that there was an antibacterial
agent there in principle. This principle later lead to medicines that could kill certain types of
disease-causing bacteria inside the body.
At the time, however, the importance of Alexander Fleming's discovery was not known. Use of
penicillin did not begin until the 1940s when Howard Florey and Ernst Chain isolated the active
ingredient and developed a powdery form of the medicine.

History of Penicillin
Originally noticed by a French medical student, Ernest Duchesne, in 1896. Penicillin was rediscovered by bacteriologist Alexander Fleming working at St. Mary's Hospital in London in
1928. He observed that a plate culture of Staphylococcus had been contaminated by a blue-green
mold and that colonies of bacteria adjacent to the mold were being dissolved. Curious, Alexander
Fleming grew the mold in a pure culture and found that it produced a substance that killed a
number of disease-causing bacteria.
Naming the substance penicillin, Dr. Fleming in 1929 published the results of his investigations,
noting that his discovery might have therapeutic value if it could be produced in quantity.

Dorothy Crowfoot Hodgkin

Hodgkin used x-rays to find the structural layouts of atoms and the overall molecular shape of
over 100 molecules including penicillin.
Dorothy's discovery of the molecular layout of penicillin helped lead scientists to develop other
antibiotics.

Dr. Howard Florey

It was not until 1939 that Dr. Howard Florey, a future Nobel Laureate, and three colleagues at
Oxford University began intensive research and were able to demonstrate penicillin's ability to
kill infectious bacteria. As the war with Germany continued to drain industrial and government
resources, the British scientists could not produce the quantities of penicillin needed for clinical
trials on humans and turned to the United States for help. They were quickly referred to the

Peoria Lab where scientists were already working on fermentation methods to increase the
growth rate of fungal cultures. One July 9, 1941, Howard Florey and Norman Heatley, Oxford
University Scientists came to the U.S. with a small but valuable package containing a small
amount of penicillin to begin work.
Pumping air into deep vats containing corn steep liquor (a non-alcoholic by-product of the wet
milling process) and the addition of other key ingredients was shown to produce faster growth
and larger amounts of penicillin than the previous surface-growth method. Ironically, after a
worldwide search, it was a strain of penicillin from a moldy cantaloupe in a Peoria market that
was found and improved to produce the largest amount of penicillin when grown in the deep vat,
submerged conditions.

Andrew J. Moyer
By November 26, 1941, Andrew J. Moyer, the lab's expert on the nutrition of molds, had
succeeded, with the assistance of Dr. Heatley, in increasing the yields of penicillin 10 times. In
1943, the required clinical trials were performed and penicillin was shown to be the most
effective antibacterial agent to date. Penicillin production was quickly scaled up and available in
quantity to treat Allied soldiers wounded on D-Day. As production was increased, the price
dropped from nearly priceless in 1940, to $20 per dose in July 1943, to $0.55 per dose by 1946.
As a result of their work, two members of the British group were awarded the Nobel Prize. Dr.
Andrew J. Moyer from the Peoria Lab was inducted into the Inventors Hall of Fame and both the
British and Peoria Laboratories were designated as International Historic Chemical Landmarks.

Andrew J Moyer Patent

On May 25, 1948, Andrew J Moyer was granted a patent for a method of the mass production of
penicillin.

Resistance to Penicillin
Four years after drug companies began mass-producing penicillin in 1943, microbes began
appearing that could resist it.
The first bug to battle penicillin was Staphylococcus aureus. This bacterium is often a harmless
passenger in the human body, but it can cause illness, such as pneumonia or toxic shock
syndrome, when it overgrows or produces a toxin.

Penicillin history: The discovery of penicillin, one of the major events in the history of
medicine.
In 1871 the scientist Joseph Lister by chance noticed that the mould which grows on
cheese and fruit can make microbes (germs) grow weaker. He made some successful
experiments on patients but did not, it seems, fully recognize the implications of his
findings.
In 1906 the young physician Alexander Fleming became a research assistant at St
Mary's Hospital in London. Fleming went to France during the First World War to treat
wounded soldiers and could see for himself that there was no effective way of treating
many infections. Back at St Mary's after the war, Fleming was determined to find a
better way of killing germs. In 1928 he was studying staphylococci bacteria (that can,
among other things, infect wounds). By pure luck, he noticed that on a dish containing
agar on which he had been growing germs, near some mould, the germs were less
common. He grew more of the mould, naming it penicillin from its Latin name
Penicillium.
Fleming discovered the mould was effective against bacteria that caused diseases such
as anthrax, meningitis and diphtheria. He published his discoveries but did not have the
resources to experiment more widely with penicillin.
In 1938 Howard Florey, professor of pathology at Oxford University, started work on
some of the mould that Fleming had grown. Florey was an Australian doctor who had
already made a number of important discoveries and was also studying natural
substances that could kill bacteria. He had been impressed by an article that Fleming
had published in 1929 describing the effects of penicillin. Unlike Fleming, Florey had a
large research department, including Ernst Chain, a German Jew who had fled the Nazis.
By 1939 Florey and Chain had begun to realize the importance of penicillin and they had
saved a number of people's lives with it as of 1942.
The widespread use of penicillin was speeded up by the advent of World War II. The
governments of the US and Britain resolved to produce all the penicillin they could and
by then, the manufacture of other drugs had become big business, so many major
companies were eager to produce penicillin. In 1943 Florey went to Russia to help it
make penicillin and by 1944, there was enough penicillin for all the allied troops
wounded in the D-Day invasion of Europe.

1945 Chain, Fleming and Florey shared the Nobel Prize in Physiology or Medicine "for
their discovery of penicillin and its curative effect in various infectious diseases." That is
where the usual story of penicillin stops.
The name of Norman G. Heatley (1911-2004) is rarely mentioned in this context. An
Oxford University biochemist, Dr. Heatley developed a technique for isolating penicillin
during World War II but it only yielded minuscule amounts of the precious substance.
To make more penicillin, Dr. Heatley had to assemble an apparatus from bottles,
containers and hospital bedpans Against enormous odds, Dr. Heatley's machine worked.
It separated enough penicillin to test on a human, but the limited supply ran out before
the patient could fully recover.
In June 1941, Drs. Heatley and Florey went with samples of their penicillin mold to a
Department of Agriculture research laboratory in Peoria, Illinois, where, working with
some of the top biologists in the US, they set in motion the research that led to largescale production of penicillin.
"Not only did Heatley make a crucial contribution, but without him the enterprise may
not have succeeded at all," said Sir James Gowans, a former professor of pathology at
Oxford. "He was the one that came up with the key step in the isolation of penicillin." Dr.
Heatley, however, was not included in the Nobel Prize Physiology or Medicine for the
development of penicillin. Nobel Prizes honor great contributors but they often omit
great contributors.

Penicillin was discovered by Alexander Fleming in 1928, and in 1940, several years before the
introduction of penicillin as a therapeutic, a bacterial penicillinase was identified by two members
of the penicillin discovery team (1). Once the antibiotic was used widely, resistant strains capable
of inactivating the drug became prevalent, and synthetic studies were undertaken to modify
penicillin chemically to prevent cleavage by penicillinases (-lactamases). Interestingly, the
identification of a bacterial penicillinase before the use of the antibiotic can now be appreciated
in the light of recent findings that a large number of antibiotic r genes are components of natural
microbial populations
Following the discovery of penicillin, it seemed that S. aureus infections were controllable;
however, the respite from resistance was short-lived. The landmark discovery and introduction of
methicillin (the first designer antiresistance antibiotic) in 1959 were thought to be a sure defense
against the penicillinases, but the appearance of methicillin-resistant S. aureus (MRSA) within just
3 years led inexorably to other multiantibiotic-resistant variants, and the acronym now denotes
multidrug-resistant S. aureus. Relatively recently, MRSA has moved outside the hospital and
become a major community-acquired (CA) pathogen, with enhanced virulence and transmission
characteristics. CA-MRSA has most of the properties of MRSA, albeit with different mec gene
clusters, and has acquired new pathogenicity genes, such as the gene encoding the cytotoxic
Panton-Valentine leukocidin (44). These are regulated by defined signaling systems (101).
A long-recognized hospital denizen, the toxin-producing anaerobe Clostridium difficile, is
increasingly found as the cause of severe intestinal infections; recently, hypervirulent toxin-

producing strains have been recognized (80, 145). Being a Gram-positive spore former, it is a
hardy organism and is readily transmitted by hospital personnel, on equipment, and as aerosols.
Its renewed prominence is considered the result of extensive hospital use of antibiotics such as
expanded-spectrum cephalosporins, the newer penicillins, and fluoroquinolones that cause
significant depletion of the Gram-negative intestinal microflora, thus enhancing C. difficile
colonization. In other words, these infections are the direct result of antibiotic use.

Penicillin: The Turning Point

By: Camille Adames
Samantha Victores
Jenny Shim
Thesis Statement: The discovery and creation of penicillin is a turning
point in history because it increased life expectancy, broadened medical
research and practice, and created healthier sanitary conditions.
Life in the 1940's centered around World War II. Men were going off to
war and women were doing the men's jobs, They did what they could
have not done when the men were there, such as play a professional
sports or do jobs that men usually took . Many soldiers suffered from
war wounds and often died from them. Some were lucky to survive long
enough to see their family, friends, and spouses; but they died
eventually from the infections. Of course, this could have happened to
anyone in the 1940's or before. A simple cut from knife could lead to
your death because there were no antibiotics to stop bacteria from
growing and entering your body. Due to this, people were not really
expected to live past the age of 60 or 68. People could have also died
from surgeries. People who had surgeries had a higher chance
of getting an infection than those of us today because, again, there was
nothing to stop bacteria to grow. People began to collect scrap metals to
help the war effort, especially after the tragic bombing of Pearl
Harbor. The center of everyone's life in the world was World War II, and
people all over the globe were affected by the outcomes of the war.

History of Penicillin

Penicillin was accidentally invented by Alexander

Fleming. Around August or September of 1928 Sir
Alexander Fleming started growing staphylococcus
bacteria in a culture plate. He left it untouched for a while
and he had accidentally let it grow mold. He noticed that
the mold had been killing the bacteria. Instead of throwing
out the bacteria like any other person would, he began to
study the mold and bacteria. Fleming realized that the
mold is called Penicillium notatum, hence the name
penicillin. At first, Fleming thought of this discovery as
useless because it was very hard to produce, meaning it
can not be easily mass produced to be distributed among
people. He eventually gave up, but his work was later
used to create the first mass produced antibiotic.

Turning Point in History

To Economic Impact

In the late 1930's, Howard Florey and Ernst Chain found

Flemming's research and helped to develop penicillin.
They discovered an easier way to purify penicillin. When
Florey and Chain first tested penicillin, they did not use
humans. They actually used eight mice. Four of the mice
were given penicillin and four were not. Six mice died and

two given the medicine survived. Then, Florey and Chain

tested penicillin on a human.On February 1, 1941, a
policeman with a severe case of staphylococcus was
given the antibiotic. The penicillin ran out and the man
died, but he did get much better over the time he had
penicillin. Chain and Florey soon discovered that the corn
steep liquor could be used to mass produce penicillin. On
top of that Ernest Chain and Howard Florey convinced a
big company to produce and sell penicillin. This event plus
the discovery of using corn steep liquid really helped to
launch the reign and mass impacts of penicillin. This was
the era of the "wonder drug." Penicillin was a huge
accomplishment and saved many lives.

Economic Impact
To Political Impact

The United States economy has been through many

ups and downs because of
penicillin. In the 1940s when penicillin was first produced,
it helped fight many infections
and diseases, such as syphillis. Also, penicillin was very
important for big operations.
These caused penicillin to be in high demand. Distributers
of penicillin gained a lot of
money from hospitals, benefiting the economy. However,
before penicillin was produced,
many people had to be treated for minor things. This put
economic strain on hospitals
and the sick person's family. Men who were sick or had
passed away due to illnesses

caused less workers. This lead to bad economy.

Since penicillin needed to be produced at high rates,
a lot of money was needed,
and the economy at that time was not very good. The
money used for penicillin could
harm the economy even more. However, the penicillin
produced was sold and a lot of
money has been made off of that. Other antibiotics made
after penicillin also contributes
to the money made. If antibiotics were never made, people
would have to spend billions
more on just on healthcare. After penicillin, the cost of
penicillin was brought down. When
penicillin was first produced, it was more expensive. It
used to cost a few dollars for 100,000 units of penicillin,
but now it only costs a few cents for the same amount of
penicillin. This could allow the government put those few
dollars into a fund for something else.

Political Impact
Button Text

Over the years, penicillin has taken a toll on the

government. The governments
job is to help save biodiversity and the environment.

Biodiversity is what we use for

the production of penicillin. The Government has taken a
lot of time, money and
effort to help penicillin. First of all, the government must
spend a lot of money to fund
the research and production of antibiotics, including
penicillin. However, the production
has slowed down a lot, so the government did not have to
as much money than before.
Many national governments must create laws and
guidelines for the distribution of penicillin, for they want to
finance facilities in order to encourage the development of
antibiotics.
Governments must be in charge of preserving effective
antibiotics and the research and development of new
ones. Research takes place in government agencies, and
they must
pay for them as well. Even today research for penicillin still
continues.
Some committees are established for infectious
diseases. For example,
one committee, named the Swann Committee, wants for
penicillin and tetracycline
to be banned as growth promotes in the United Kingdom.
However, these temporary
expert panels concerned with infectious diseases were
soon replaced by standing advisory committees. Penicillin
is now under the control of the Therapeutic Substances
Act. The U.S. government has worked with other foreign
government for the ownership of penicillin. This has gave
lots of fame to the United States.

Penicillin was a big help during World War II. During

the war, many soldiers got
wounds and illnesses, therefore penicillin contributed in
saving lives. This caused more
soldiers to fight, which meant a greater chance that the US
would not lose this war and
getting attacked. This caused good outcomes of the war
and benefits the governments.
Also, if penicillin wasn't used in past wars, we could have
lost the war due to less soldiers
and been overrun by foreign governments and many more
other things. Now, the government would not have to
forcefully choose someone to join the army.

Penicillin: An accidental discovery changed the course of medicine

Penicillin was first discovered in 1928 and is now the most widely
used antibiotic in the world.
It all started with a mold that developed on a staphylococcus culture plate. Since then, the
discovery of penicillin changed the course of medicine and has enabled physicians to treat
formerly severe and life-threatening illnesses such as bacterial endocarditis, meningitis,
pneumococcal pneumonia, gonorrhea and syphilis.
Penicillin discovered
Sir Alexander Fleming, a Scottish researcher, is credited with the discovery of penicillin in
1928. At the time, Fleming was experimenting with the influenza virus in the Laboratory of
the Inoculation Department at St. Marys Hospital in London.
Often described as a careless lab technician, Fleming returned from a two-week vacation to
find that a mold had developed on an accidentally contaminated staphylococcus culture
plate. Upon examination of the mold, he noticed that the culture prevented the growth of
staphylococci.
An article published by Fleming in the British Journal of Experimental Pathology in 1929
reads, The staphylococcus colonies became transparent and were obviously undergoing

lysis the broth in which the mold had been grown at room temperature for one to two
weeks had acquired marked inhibitory, bactericidal and bacteriolytic properties to many of
the more common pathogenic bacteria.
Fleming described
rapidly increases
and changes color

the colony as a fluffy white mass which

in size and after a few days sporulates
from dark green to black to bright yellow.

Even in the early

effect against
effective against

experimentation stages, penicillin had no

gram-negative organisms but was
gram-positive bacteria.

Published reports
sometimes finds
up just after dawn
to revolutionize all
antibiotic, or
exactly what I

credit Fleming as saying: One

what one is not looking for. When I woke
on Sept. 28, 1928, I certainly didnt plan
medicine by discovering the worlds first
bacteria killer. But I guess that was
did.

Though Fleming
stopped studying penicillin in 1931, his
research was
continued and finished by Howard Flory
and Ernst Chain,
researchers at University of Oxford who
are credited with the development of penicillin for use as a medicine in mice.
Widespread use
Penicillin made a difference during the first half of the 20th century. The first patient was
successfully treated for streptococcal septicemia in the United States in 1942. However,
supply was limited and demand was high in the early days of penicillin.
Penicillin helped reduce the number of deaths and amputations of troops during World War
II. According to records, there were only 400 million units of penicillin available during the
first five months of 1943; by the time World War II ended, U.S. companies were making 650
billion units a month.
To date, penicillin has become the most widely used antibiotic in the world. by Katie
Kalvaitis

How penicillin changed medicine: perspective from an infectious disease expert

The discovery of penicillin changed the world of medicine enormously. With

its development, infections that were previously severe and often fatal, like
bacterial endocarditis, bacterial meningitis and pneumococcal pneumonia,
could be easily treated. Even dating all the way back to World War II and
today with the war in Iraq, soldiers experienced injuries that would have
been fatal without penicillin and other antibiotics that were developed
subsequently. It is really impossible for me to imagine what the world would
be like without penicillin. I question whether there would be a discipline of
infectious diseases as we know it today.

Theodore C.
Eickhoff

There were beginning treatments for pneumococcal pneumonia in the 1930s with antisera
and sulfonamides, but use of these treatments quickly came to a halt, and everyone began
using penicillin. This quickly led to a number of pharmaceutical industries beginning to
screen a variety of other natural products for antibacterial activity, which led to a whole host
of new antibiotics, such as streptomycin, aminoglycosides, tetracycline and the like.
Penicillin clearly led the way in that development.
It is interesting that using penicillin for the treatment of infections like pneumococcal
pneumonia and bacterial endocarditis never had a randomized, controlled trial because the
difference with treatment was so clearly apparent that no one even thought of doing a
randomized controlled trial.

Penicillin was discovered when Sir Alexander Fleming noticed that Penicillium notatum could
neutralize a bacterial colony of Staphylococcus aureus, according to About.com. Fleming made
the discovery by mistake when he left a plate culture of the bacteria open for mold contamination
unintentionally, claims Wikipedia.
French scientist Ernest Duchesne studied the link between mold and bacteria in 1897, but his
research was ignored, notes Wikipedia. He demonstrated his research by curing guinea pigs that
were suffering from typhoid.
Alexander Fleming was a bacteriologist who worked at St. Mary's Hospital in London, notes
About.com. His research revealed that the antibacterial properties of the bluish-green mold could
be used to eliminate bacteria that caused certain ailments.
Fleming grew the mold and discovered that it released a substance that neutralized a variety of
diseases, reports About.com. However, Fleming's research was not deemed noteworthy at the
time. He named the mold penicillin, and he published his research in 1929, believing that it could
be an effective medicine if produced on a mass scale.
It was not until 1939 that Dr. Howard Florey and three other colleagues at Oxford University
effectively proved that penicillin could kill bacteria, explains About.com. Since the scientists

could not conduct clinical trials because of the war effort, they concluded their research in the
United States. Penicillin was produced on a commercial scale in the early 1940s, and it was used
to treat soldiers who fought on D-Day.

Penicillin: the first miracle drug

Many of you are here only because penicillin

saved your life, or the life of one of your
parents or grandparents. Penicillin's ability to
cure people of many once-fatal bacterial
infections has saved so many lives that it is
Early penicillin culture facility at the Sir
easy to understand why it was once called a
William Dunn School of Pathology,
"miracle drug".
Oxford, England.
Museum of the History of Science,
Antibiotics are chemicals, effective at very low
Oxford

concentrations, created as part of the life

process of one organism, which can kill or
stop the growth of a disease-causing microbe--a germ. In 1929, Alexander Fleming, a doctor
and researcher at St. Mary's Hospital in London, England, published a paper on a
chemical he called "penicillin", which he had isolated from from a mold, Penicillium
notatum. Penicillin, Fleming wrote, had prevented the growth of a neighboring colony of
germs in the same petri dish. Dr. Fleming was never able to purify his samples of penicillin,
but he became the first person to publish the news of its germ-killing power. Howard
Florey, Ernst Chain and Norman Heatley expanded on Fleming's work in 1938, at Oxford
University. They and their staff developed methods for growing, extracting and purifying
enough penicillin to prove its value as a drug.
World War II (1939-1945) had begun by the time their research was showing results. The
main research and production was moved to the United States in 1941, to protect it from
the bombs pounding England. Work began on how to grow the mold efficiently to make
penicillin in the large quantities that would be needed for thousands of soldiers. As the
destruction of the war grew, so did interest in penicillin in laboratories, universities and
drug companies on both sides of the Atlantic. The scientists knew they were in a race
against death, because an infection was as likely to kill a wounded soldier as his wound.

Photograp
h courtesy
of
Associate
d Press

Petri dish
containin
g colony
of
Penicilliu
m
notatum.
Sold by
Christie's
Auction
House
in 1998
for
$13,121.

Chains of
conidia
(spores)
produced by
hyphal branch
from
mycelium

Photograph courtesy of Associated Press

Sir Alexander Fleming, 1952

Creating the right environment for growth was the first step in producing enough penicillin
to be used as a drug. In Oxford, experiments showed that Penicillium notatum grew best in
small shallow containers on a broth of nutrients. Penicillium need lots of air. In the United
States, it was discovered that huge "deep fermentation" tanks could be used if sterilized air
was pumped continually through the tanks. Production increased even more when corn
steep liquor, a thick, sticky by-product of corn processing, was added to the tanks. Corn
steep liquor contained concentrated nutrients that increased the yield 12-20 times.
Formerly considered a waste material, corn steep liquor became a crucial ingredient in the
large-scale production of penicillin.
Scientists were also determined to find another strain of Penicillium that might grow better
in the huge deep fermentation tanks. Army pilots sent back soil samples from all over the
world to be tested for molds. Residents of Peoria, Illinois, were encouraged to bring moldy
household objects to the local U.S. Department of Agriculture laboratory, where penicillin
research was being conducted. Laboratory staff members also kept an eye out for
promising molds while grocery shopping or cleaning out their refrigerators.

Photograph courtesy of Merck

Archives, Merck & Co. Inc.
Photograph courtesy of
Photograph courtesy of Upper part of fermentors
Merck Archives, Merck
Merck Archives, Merck (tanks) used to produce
& Co. Inc.
& Co. Inc.
penicillin and vitamin B12.

Refrigeration
equipment for large
fermentation unit at
Cherokee Plant,
Danville, PA.

Fermentation unit
used in purifying
penicillin in 1945.

In 1943, laboratory worker Mary Hunt brought in an ordinary supermarket cantaloupe

infected with a mold that had "a pretty, golden look." This Penicillium species, Penicillium
chrysogenum grew so well in a tank that it more than doubled the amount of penicillin
produced. The deep fermentation method, the use of corn steep liquor and the discovery of
P. chrysogenumby Mary Hunt made the commercial production of penicillin possible.
Researchers continued to find higher-yielding Penicillium molds, and also produced higher
yielding strains by exposing molds to x-rays or ultraviolet light.
Penicillin kills by preventing some bacteria from forming new cell walls. One by one, the
bacteria die because they cannot complete the process of division that produces two new
"daughter" bacteria from a single "parent" bacterium. The new cell wall that needs to be
made to separate the "daughters" is never formed.
Some bacteria are able to resist the action of antibiotic drugs, including penicillin.
Antibiotic resistance occurs because not all bacteria of the same species are alike, just as
people in your own family are not exactly alike. Eventually, the small differences among the
bacteria often mean that some will be able to resist the attack of an antibiotic. If the sick
person's own defenses can not kill off these resistant bacteria, they will multiply. This
antibiotic-resistant form of a disease can re-infect the patient, or be passed on to another
person.

Taking antibiotics for viral illnesses like colds can also cause antibiotic resistant bacteria to
develop. Antibiotics have no effect on viruses, but it will kill off harmless and even the
beneficial bacteria living in the patient's body. The surviving resistant bacteria, free from
competition, will live and multiply and may eventually cause disease.
Patients with bacterial infections, who don't finish their antibiotic prescriptions completely,
also allow resistant bacteria to develop. This happens because a small number of semiresistant bacteria, which needed the full course of antibiotics to kill them, survive. Instead
of being a small part of the bacteria causing an infection, the more resistant bacteria take
over when sensitive bacteria are killed by the antibiotic.
Today, in the United States, deaths by infectious bacterial diseases are only one-twentieth of
what they were in 1900, before any antibiotic chemicals had been discovered. The main
causes of death today are what are referred to as "the diseases of old age": heart disease,
kidney disease and cancer. We would be shocked to hear of someone dying from an
infection that started in a scratch, but, before antibiotics like penicillin, it was common for
people to die from such infections.
Humans can slow the creation of antibiotic resistant diseases by understanding the uses and
limits of antibiotics. Take all of an antibiotic, and only take them when prescribed by a
doctor. Research to develop new antibiotics to treat resistant bacteria continues, but
research takes time. Time is running out because the world's biodiversity is decreasing--the
source of half of our disease-fighting chemicals.
An example of the importance of preserving the world's biodiversity occurred in 1996, in
New York state. Students at Cornell University collected a fungus that finally made it
possible to identify the two very different life stages of the mold that produces the drug
cyclosporin. Cyclosporin prevents the rejection of transplanted organs. Without it,
transplant operations would be impossible. Knowing the full life history of the cyclosporinproducing fungus may make it easier to find related molds. Even people who see no special
beauty or value in the world's biodiversity may one day benefit from the currently
unknown and powerful substances, produced by fungi and other microbes, that are waiting
for discovery in familiar places.

Penicillin: Who Found This Functional Fungus

The first name for penicillin was mould juice. Scottish bacteriologist Alexander Fleming
accidentally discovered the antibiotic in 1928, when he came back from a vacation and found
that a green mold called Pennicilium notatum had contaminated Petri dishes in his lab and
were killing some of the bacteria hed been growing.

So he isolated the mold, grew more of it, and then experimented to see how many other bacteria
it could kill. Lots of them, it turned out. We now know that penicillin works by preventing
bacteria from forming new cell walls. No new walls, no new cells, no new bacterial growth.
If you lived back then, almost a century ago, you could die from a scratch if it got infected. Or a
dental procedure. Or a deep cut. Or any of the many, many things that can happen, whether
youre working in the backyard, making dinner, or fighting in battle.
And thats partly why Fleming was so keen on finding a way to control infections. As a captain
in the Royal Medical Corps during World War I, hed worked in battlefield hospitals in France,
where soldiers died from infected wounds. So he made it his goal to find antibacterial
substances.
When he did find P. notatum, he didnt formally name it penicillin until March 7, 1929. He
wasnt able to produce enough to help all the people who needed it, but he did publish his
research.
In 1938, Oxford Pathologist Howard Florey discovered Flemings research and expanded on it,
working with a biochemist named Ernst Boris Chain, who had fled Germany. British biochemist
Norman Heatley further developed the work, vigorously growing and purifying penicillin.
After much research and experimentation as well as a trip to the United States, where Florey
and Heatley worked with American scientists an injectable, mass-produced form of penicillin
was ready by 1942. That was just in time to help soldiers wounded in World War II.
The numbers prove the point: Bacterial pneumonia killed 18 percent of fallen soldiers in WWI,
but in WWII it killed less than 1 percent of soldiers. Penicillin went on to change the way we
treat illness and wounds and it all started with mould juice!

Penicillin
Introduction
Today, the use of penicillin and other antibiotics are common place. The
various antibiotics are used to treat a number of what are now common
diseases and to prevent the onset of infections when our skin, our first
barrier to fight off disease, is somehow broken through a simple cut or a

more serious wound. It is something that we all take for granted, today.
However, many diseases and simple wounds that are so easily treated
today because of the availability of antibiotics has not always been
available. Antibiotics are a relatively recent discovery and the first
practical one, penicillin, was not available until the early 1940s. Even the
concept of using fungal products, such as penicillin, to produce medicine is
a relatively new one. However, many folk remedies that have included
fungi have long been utilized, but the incorporation of fungi into the
remedy was inadvertent and not known. For example, over three thousand
years ago, the Chinese had put moldy soybean curd on boils and other
types of skin infections. Other cultures have placed warm earth, which
contains molds and other fungi, as first aid in injuries. There was
undoubtedly antibiotics in the soybean curd and earth that were placed on
injuries. So, although the concept of antibodies is relatively recent, its
use has been around for some time.
The discovery of penicillin has often been described as a miracle drug,
and that is exactly what it was. Prior to the discovery of penicillin, death
could occur in what would seem, today, to be very trivial injuries and
diseases. It could occur from minor wounds that became infected or from
diseases such as Strep Throat, and venereal diseases such as syphilis and
gonorrhea were a much more serious issue.
Early in 2005, an email came asking for information about penicillin. The
email was from Dr. Morton Paterson, a retired philosophy professor, now
living in Canada. He was writing his autobiography for his grandchildren as
a legacy for them. Part of that autobiography had to do with the impact
penicillin played in his life. When he emailed me the story, I knew that it
was one that I wanted to share. As a boy, just prior to the discovery of
mass producing penicillin, Dr. Paterson had badly scraped his knee, an
injury that he almost died from. The following is his account of this injury
and how it was treated at that time:

It was the late spring of 1942, and I was seven years old. My sister Lorna had
just been born. One day I was outside playing with my friends - running while playing
tag or something. There weren't any parks or grassy fields, so the kids played on the
rocks or on the streets. I fell on the street, which was covered with chunks of slag
(waste from the Smelter), and scraped my right knee. I guess it was bleeding pretty
bad, so I ran home. Later I was told that it was on a Wednesday, and that my
temperature shot up and up. By Saturday Mum and Dad had a sick boy on their
hands, so on the advice of Dr. Chappell, our family doctor, I was rushed to St.
Joseph's Hospital in Sudbury.
The cut on my knee had become infected, and I had blood poison. For a few days
I guess I was "out of it", in a coma, and hung in the balance between life and death. I
was diagnosed as having osteomyelitis, which means "bone infection". Apparently what
happens with osteomyelitis is that the infected blood seeks out a part of the body
which is already weak for some reason. In my case that happened to be the socket in
my left hip.
Don't ask me why I had a weak hip - cause I don't know the answer. I just did.
Anyway, they knew they had to operate fast to stop the infection before it
traveled to a vital organ. That led to three months in hospital. The surgeon was Dr.
Mowat, and I remember him as a very kind and soft-spoken man. He had to scrape out
the infected bone, but then leave the large incision open so the nurses could pack it
every day with fresh gauze. Later I was told that the reason for not closing up the
incision was that oxygen (fresh air) was needed to clear up the infection. Without
oxygen the infection would stay in the bone, and be a continuing threat.
I've never been so scared in all my life. I didn't know why my hip was so sore
and not getting better, and could tell that Dr. Mowat and my parents were pretty
worried. As the nurses peeled away the old packing and re-packed my hip with fresh
gauze they tried their best to cheer me up and not let on they were worried. I
remember them saying, "Now be a brave little soldier, Mortie!"
Surgery had to be performed a few more times to clean out bone chips in the
incision. All I can remember about those extra surgeries was being wheeled out of my
room, down the corridor, and into a large bright "operating room". Suddenly a doctor
(I later learned he or she is called the anesthetist) behind me would cover my face
with a cloth and tell me start counting. Then the doctor would a couple of drops of
ether onto the cloth. I would get to about 3 before falling asleep.
Looking back to that operating room experience these sixty-three years later I
still remember my panic, crying out when the cloth went over my face. Ether had the

most sickening smell I ever smelled, and I guess the scariest part was not knowing
when they'd cart me down the corridor again and have that awful cloth suddenly
draped over my face. Another thing about ether was that I'd be so sick when I came
to back in my room. The smell seemed to, linger forever, and I kept bringing up. The
nurses would give me a pill to help me sleep, so eventually I'd doze off.
Nowadays children who are going to have surgery are given a tour of the
operating room ahead of time, and told quite a bit about what's going to happen. Also,
ether isn't used any more, and I think the anesthetic is usually given with a needle.
That's about it. When the infection was finally contained (by mid-summer), less
and less packing was put into the incision till the day finally came that I could go
home on little crutches that I still have. But there are a couple of other things I
want to say about that summer.
One evening about ten o'clock, shortly after I'd been admitted to hospital, two
nurses came to my bedside to check up on me. I wasn't asleep yet, but my eyes were
closed and they thought I was. I remember clearly one of the nurses saying to the
other, "Do you think he'll make it?" The other nurse answered, "I doubt it."
I didn't let on that I'd heard them. I was likely scared enough already so that
the extra dose of pessimism didn't really register. Or - and this is quite likely true -

perhaps little Mortie made up his mind right then and there that he WAS going to
make it.
I wouldn't have put it this way back then, but a line made famous forty years later by
Prime Minister Pierre Trudeau may have been forming behind those closed eyes; " Just

watch me!"

So after several months and with a lot of self determination, "Mortie"

was finally able to go home. At first he was only able to walk with
crutches, but was eventually able to walk without them. However, to this
day he still walks with a limp from that accident so many years ago. In
1946, there was a reoccurrence of the osteomyelitis, but by this time
penicillin was available to the general public. The following, again, has been
written by Dr. Paterson:
In 1946 my osteomyelitis returned!
Darn it. We had all thought that it was over and done with, even though we'd been
warned that it might come back. But this time the infection was gone in only a few days.

Now he was working underground at Frood Mine. The income from selling insurance just
didn't pay the everyday bills for a family of five, plus now the extra cost of Morty's
hospitalization.
This was twenty years before universal medical coverage (Medicare) in Canada, so anybody
not covered by a private plan had to pay their own medical bills. In 1942 Dad had worked
for INCo, so the St. Jo's Hospital costs were covered. This time hospital costs (at INCo's
private Copper Cliff Hospital) would be covered again by the INCo plan. I was there for
only for a short time, not months.
Frood was the deepest of all the mines in the Sudbury area (some said the main
shaft was a mile deep!), and Dad worked down there for three years. Mining is always
dangerous, but so-called "hard rock" mining (for nickel) is safer than "soft-rock" mining
(for coal). Many Nova Scotia "men of the deep" have died in coal mines, but few nickelminers. Still, the danger is always here, mostly from dynamite blasts that go wrong.

So Dad chose a hazardous job so his son could get hospital care, He worked
days and afternoon shifts, and at times "bonus hours" as well. But Mum couldn't
sleep when Dad was on afternoon shift. and getting home around 1 am or later.
There was lots of stress at home.

Penicillin was a miracle drug that was now available for the treatment of conditions

caused by infection - like osteomyelitis. Sulfa drugs had stopped the bone disease in '42,
but still a hip fusion was needed. Also, though the sulfa worked on the short run, it did
not eliminate the infection from the bone marrow.
This time Dr. Mowat used penicillin to stop the infection right away. It also
eradicated it completely. Not once in all these years did osteomyelitis ever come back!
Miracle drug, indeed yes. But what is truly amazing is that penicillin was eventually
produced in large quantities in order to treat soldiers who were dying on the battlefields
of Europe - not directly because of their wounds, but because of poisoned blood that
carried the infection that set into those wounds. When they were treated with penicillin,
many soldiers recovered. Of the thousands of soldiers who had died in battle in prepenicillin World War I, many had died from pneumonia from infection.
Remember that a nurse had told me in 1942 to "be a good little soldier"? I wasn't a

real soldier, but I'm glad she said that.

So Dr. Paterson required medical care twice for osteomyelitis; before and
after penicillin became available. From reading his story, I think we can all
appreciate the importance of penicillin.

Dr. Paterson was not the only one to have osteomyelitis. Mickey Mantle,
the former New York Yankee, centerfielder, Hall of Famer, also
contracted this disease while in high school. While playing high school
football, Mantle was kicked hard in his right shin. His ankle became
swollen to twice its normal size and he later had a 104 F. He eventually
developed osteomyelitis. When he was finally taken to the hospital, the
doctors thought the only thing that could be done was to amputate his
leg. However, his mother would have none of that and took him to another
hospital where the doctors treated him with penicillin. The penicillin
saved his leg and he went on to have a successful baseball career.
The Story of the Discovery of Penicillin
You are all undoubtedly familiar with the story of penicillin. In all
introductory text books, in the life science, the story always tells how
penicillin was discovered accidentally, at St. Mary's Hospital, in London,
by Dr. Alexander Fleming. Fleming was examining a culture of

Staphylococcus aureus, a pathogenic bacterium on which he was doing

some research, when he noticed that it had become contaminated by a
species of Penicillium. Although, the species of the mold was unknown to
Fleming, at the time, he did observe that it was inhibiting the bacterial
growth. Fleming wrote a paper on his findings in 1929 and the rest is
history. However, it was never that simple. Such a short summary really
does not tell you the entire story, and in this case, says that Fleming's
discovery of penicillin was one of chance and does not credit other people,
who were just as deserving or more so in the development of penicillin for
medicinal use.
Some luck, surely was involved, as is true with many events. Alexander
Fleming did not have any ambition to become a doctor throughout his life.
His start into bacteriological medicine came from an unlikely string of
events. In 1900, when the Boer War broke out between England and

colonies in southern Africa, Fleming and two brothers joined a Scottish

regiment, which turned out not to be such a dangerous time for them as
they had chanced upon a country club environment. They spent much of
their time shooting, swimming, and even playing water polo. Following the
war, Fleming returned home to discover that his uncle had died and left
him and his brothers with a sizable inheritance. His older brother, Tom,
who was a successful doctor by this time, advised him to invest his money
on his career and suggested that he attend medical school. Fleming
scored high on his examination and was able to select from three medical
schools. He knew nothing of these schools and selected St. Mary's
Hospital, in London, only because he had once played water polo against
them. After graduation, Fleming had trained to be a surgeon for just as
random a reason, but then found himself in a choice that was even more
bizarre. He had the option of taking a position, as a surgeon and leaving
St. Mary's or he could join the Inoculation Service and stay at St.
Mary's. The major influence on Fleming staying was that the captain of
St. Mary's rifle club knew of his option and was desperate to improve his
team. Knowing that Fleming was a great shot he did all he could to keep
him at St. Mary's. He convinced Fleming to join his department in order
to work with its brilliant director -- and to join the rifle club. Fleming
would stay at St. Mary's, where his discovery of penicillin was made, and
for the rest of his career .

The Discovery of Penicillin

That Alexander Fleming discovered penicillin by chance is a myth. Before
Fleming, there were a series of observations that influenced his research,
and allowed him to come to the correct conclusion when a chance
contamination in his bacterial culture was observed. This same
opportunity came to others as well, but their only response to the

contamination was that it had ruined their experiment and they had
discarded the cultures and thought nothing more of it.
Discoveries in science are rarely made by chance. Often, it involves
knowledge that has been gained over a long period of time so that all
discoveries, today, have come about because we have "stood on the
shoulders of giants that have come before us." Another words,
discoveries are based on previous discoveries from the past. In the case
of of penicillin, it was based upon historical records that had originated
as early as 1500 B.C. There were already records describing the use of
molds and fermented materials in the treatment of diseases. Because
such treatments were carried out without an understanding on the nature
of the cure or an understanding of cellular and biochemical processes of
the human body, there was a likelihood of the patient dying from the
treatment as well as being cured. It would not be until the late nineteenth
century when Pasteur put forth the concept of the Germ Theory of
Disease, i.e., diseases were caused by microorganisms. Not until then was
there any concerted effort made that would destroy the microorganisms
that were responsible for the actual causes of diseases. This led to the
search for "The Magic Bullet". Another words a search was under way to
find something that could kill the disease causing organisms without
harming the person that it was infecting. One of the most common
problems that occurred, and still occurs today, was the contamination of
bacterial cultures by other microorganisms, especially fungi. These
contaminations led to a number of observations in the late 1800s:

Joseph Lister, an English surgeon observed, in 1871, that urine

samples contaminated with mold did not allow the growth of
bacteria.

In 1874, William Roberts observed that bacterial contamination is

generally absent in cultures of the mold Penicillium glaucum.

Louis Pasteur and Jules Francois Joubert, in 1877, observed that

cultures of the anthrax bacilli, when contaminated with unidentified
molds, became inhibited.

In 1897, Ernest Duchesne had completed his Ph.D. on on the

evolutionary competition among microorganisms. Specifically, he
studied focusing on the interaction between E. coli and Penicillium
glaucum. Some of Duchesne observations included how the bacteria
was eliminated by the fungus when both were grown on the same
culture, and in inoculation of laboratory animals with lethal typhoid
bacilli and P. glaucum, the mold prevented the animal from
contracting typhoid. Although, he urged that further research be
carried out, he was unable to do so. Upon completing his degree, he
went into the army and while serving, died of tuberculosis.

In the 1920's, there were two unknown scientist, Andre Gratia and
Sara Dath who observed a fungal contamination in one of their
Staphylococcus aureus cultures that was inhibiting the growth of
the bacterium. They identified this as a species of Penicillium and
presented their observations as a paper. There was little attention
paid to this paper and the two scientist did not pursue this line of
research further. Perhaps it was due to the lack of interest by
other scientists who had heard their paper, or perhaps there was
not a realization as to what they had. Regardless of the reason,
these two scientists missed being a major part of the penicillin
story in not pursuing this research further.

Thus, a number of observations set the stage for Fleming's accidental

discovery. As early as 1920, Fleming was searching for antibacterial
agents, which was influenced by his war time experience. During the First
World War, Fleming witnessed the deaths of many soldiers that died not
from the wounds that were received during combat, but from septicemia
or in layman's term, blood poisoning that was responsible for the death of
the wounded soldiers following successful operations on those wounds. It

was only recently that surgeons had adopted Lister's concept of

antiseptic surgery. Lister argued that since bacterial infections was
harmful to the human body, that antiseptics should be used on the wounds
to kill the bacteria. However, Fleming while serving with a colleague, from
St. Mary's Hospital, argued that while this was true, the antiseptics also
had an adverse effect on the patient. They reasoned that the leukocytes
of the immune system was the first line of defense in preventing
infection and that application of antiseptics to the wounds of patients
would destroy the leukocytes more rapidly than the invading bacteria.
They recommended the use of a saline solution to cleanse wounds.
However, there was no one who wound follow this procedure since Lister's
antiseptic theory was at that time a revolutionary, cutting edge science
concept that experiments had proven to be true. So without a doubt,
Fleming was searching for antibacterial agents when he discovered
penicillin. In 1922, Fleming discovered lysozyme, what can be thought of
as the "little brother" of penicillin. Lysozymes are enzymes present in
biological substances as varied as egg whites, tears and mucus, that
causes bacteria to lyse or burst. The first biological substance that he
tested was mucous from his nose (he was sick with a cold at the time).
This would later be a major discovery, but at that time, lysozymes were
seen as interesting, but not with a great deal of applications since only
the less virulent bacteria would respond to these enzymes.
In 1928, he was researching the properties of the group of bacteria
known as staphylococci and became another in the long line of scientists
to benefit from a seemingly chance observation. His problem during this
research was the frequent contamination of culture plates with airborne
molds. However, he was also known as a sloppy scientist as well. Cultures
that he worked on were constantly forgotten, in his lab, which was
normally in a state of great disorder. After returning from a month long
vacation, Fleming observed that many of his culture plates were
contaminated with a fungus. He immediately threw the plates in a tray of

lysol. Fortunately, a former member of his lab was visiting and he took
the contaminated cultures that had not been submerged in the lysol to
show his visitor what he has been doing. It was only then that he noticed
the unusual inhibition zone around the fungus. He realized at this point
that that this may be something important and for the rest of that day
showed all of his colleagues the culture and continued to study the antibacterial properties of the mold. Subsequently Fleming isolated an
extract from the mold and he named it penicillin. Although his discovery
was published, there was not a great deal of attention paid to this paper.
Despite this success, further attempts by Fleming to produce a
concentrated extract of penicillin failed and he was unable to prove that
it had any therapeutic value and doubted it himself at this time.
It could be argued here that Fleming did accidentally discover penicillin
since he wasn't looking for it at that moment in time. However, he had
been looking for several years prior to this and without his background in
lysozyme research, Fleming may not have really ever considered further
investigation of the contaminating mold just as several scientists before
him had done. This was certainly the opinions of his colleagues at St.
Mary's Hospital.
As further test continued, Fleming began to realize that he was on the
verge of a great discovery. However, he still did not know the identity of
the fungus. He had virtually no background with fungi and knew little
about these organisms. Because of the lack of information on this fungus,
the press had initially called this mold "yellow magic" because when it was
cultured in a large vat of liquid nutrient, the liquid became a bright yellow
color. Although, Fleming was not very knowledgeable about fungi, he was
able to identify it as a species of Penicillium, and even gave it a tentative
species name, P. rubra, which was incorrect. However, it was his
identification to genus, that prompted Fleming to name this compound,
that inhibited bacterial growth, penicillin. The fungus would eventually be

identified by Charles Thom, who was the authority on the taxonomy of

Penicillium, as Penicillium notatum.

Although very similar to lysozymes that he had tested, it was obvious
that penicillin was far more powerful an antibacterial compound. His crude
extracts could be diluted 1,000 times and still be effective in killing
bacteria. Further test were done with various species of bacteria and also
test were carried out with laboratory animals to be certain that penicillin
would not be toxic to animals. These results were, again, very interesting,
but how could they be translated to practical use? Fleming only continued
to work on and off with with penicillin between 1928-1931, but was unable
to produce it in the quantity necessary for testing or practical
applications. In addition, there were problems in the production of
penicillin. Fleming found that, under the conditions which he was growing
his mold, penicillin was unstable and the culture stopped producing
penicillin after eight days. However, this did not dampen Fleming's
enthusiasm, he continued to work on penicillin and talk of its great
potential value in medicine. Unfortunately, this had the opposite effect on
his colleagues that he wanted. After several years of working on penicillin
and not being able to make practical use of it, many of his colleagues grew
tired of hearing his stories on what he was doing with penicillin. It was
only his conversation with Howard Florey at a professional medical
meetings that they attended that may have sparked an interest. So,
Fleming did not become famous overnight, the newspaper did not come to
interview him, representatives from pharmaceutical companies from near
and far did not come to see him when his discovery was made public, and
there were, in fact, many people that doubted that the discovery would
be of any value.
It would not be Fleming, but one of his former student, Dr. Cecil Paine,
that would be the first to demonstrate the value of penicillin in medicine.
Unfortunately, he would become the forgotten man in the development of

penicillin. Paine became inspired, to do research on penicillin, by Fleming's

original penicillin paper and not because he was inspired by Fleming in one
of his classes. Paine recalled, when interviewed about Fleming, that he
was "a shocking lecturer, the worst you could possibly imagine,'" not
because of inherent shyness, but because he never appeared enthusiastic
about his subject. This was not only Paine's opinion, but was a general
opinion of Fleming as a lecturer. This may possibly be one reason he failed
to stimulate any enthusiasm for his penicillin work among his
contemporaries.
Paine was, at that time, employed in a joint position in Sheffield, at the
University Medical School and Royal Infirmary at the time he was working
with penicillin. His first effort at using penicillin was in treating patients
with a skin infection called sycosis barbae. This infection of the hair
follicle, of the beard, is caused by Staphylococus, a very common
infection, in pre-antibiotic days. Following the infection, pus would form in
the follicle, and would become swollen and inflamed. Paine treated the
infection by soaking gauze with the crude penicillin extract that he
produced and then using the gauze to cover the patient's face. This
proved to be a failure, but did not dampen his enthusiasm to continue
working with penicillin.
Paine's next attempt, this time a dramatically successful one, at using
penicillin, involved the lacerated eye of a local miner. The stone
responsible for the laceration was still embedded in the eye and an
infection had set in which was caused by Pneumococcus, a pathogenic
bacterium whose presence normally would have required the removal of
the eye. However, after Paine irrigated the eye with the crude penicillin
extract, the miner's eye and eyesight was saved. Another case involved a
different form of eye infection. This time occurring in the eyes of a
baby. The baby had contracted gonorrhea, from his mother, during birth.
At this time, gonorrhea was not treatable and babies often became blind

through hereditary gonorrhea. Paine's irrigation of the babies eyes with

his penicillin extract was again successful.
If these successful treatments with penicillin had become known at the
time, it may have spurred the development of penicillin many years
earlier. However, Paine never published his results concerning these
patients nor did he ever present an oral paper on these patients even
though he had written and presented papers on other subjects. When
asked years later why he did not publish on this important event, he said
that he felt that since he was using a crude extract and that there was
not sufficient testing, he didn't feel it was worthy of publication.
Furthermore, he had to stop working with penicillin because he had taken
a position elsewhere which called for his research to take a different
direction. Thus, Fleming was still floundering at this time to get people to
carry out more research with penicillin. Fleming had accepted by this time
that personally, he lacked the skills to advance the science of penicillin
further. He believed now that he would at this point require the
expertise of a chemist. He would be proven right later.
As an added note on Paine. Paine would have an influence on someone who
would be important in penicillin research a few years later. While still at
Sheffield University, in 1932, he discussed his penicillin work with a newly
arrived Professor of Pathology. The professor, said Paine "took not the
slightest interest at that time." Yet six years later, he was to begin a
program of research that would lead to the mass production of penicillin.
The Professor's name was Howard Florey who we will have much more to
say about later.
By the mid 1930's, several new discoveries came about in medicine that
began to make Fleming doubt as to whether he should continue his work
with penicillin. Sulfa drugs, a class of synthetic chemical substances, was
developed in Germany and was effective in treating some bacterial

infections. These drugs inhibited the action of para-aminobenzoic acid, a

substance that bacteria need in order to reproduce. Another synthetic
drug, simply known as M and B, after the English firm of May and Baker,
where the drug was developed, was demonstrated to be effective against
pneumonia. Fleming began working with both these drugs and had stopped
working with penicillin by 1934. That he was still interested in penicillin
would, in later years, be obvious.
During this same ten year period, between 1928-1938, Dr. Howard W.
Florey, Professor of Pathology at Oxford's Sir William Dunn School of
Pathology, also became interested in research on lysozymes, and later, in
1938, in antibiotic when he came across Fleming's paper on penicillin.
Florey, however, did not work alone, he had a team of scientist that
consisted of Drs. Ernst B. Chain, Leslie Falk, Norman G. Heatley and
twenty other scientists and technicians. Where Fleming had a poorly
equipped lab with no staff support, Florey's lab was well staffed and
equipped.
There are many conflicting stories concerning the relationship between
Florey and Chain. Chain is always credited as the person who actually
began working with penicillin in Florey's lab and even located a culture
that was sub-cultured from Fleming's original isolate of P. notatum.
Florey, on the other hand, was the person in charge of the research
program, but was often thought of as not being very knowledgeable about
penicillin before Chain's rediscovery of Fleming's paper. This seems
unlikely since Florey had been on the editorial board of the journal which
published Fleming's first penicillin paper and there is every likelihood
that he may have even helped in editing it. He also knew Fleming quite well
from attending professional meetings and considering Fleming's obsession
with penicillin, it seems highly unlikely that the subject would not have
come around to penicillin. Finally, when Florey met Paine in 1932, Florey
had heard about the potential for penicillin. However, when questioned

about the latter, Florey denied that this had any influence on his
direction of research and that he had forgotten that conversation by
1938, when he choose to work on penicillin. Nevertheless, one of his
former students remembered quite distinctly that Florey mentioned
Paine's work in a lecture, at Oxford, in 1936.
One thing that Florey and Chain did agree upon was that scientific
interest rather than the desire to introduce a life-saving drug was the
reason they devoted their efforts into penicillin research. This would be
one of the few things that Chain and Florey would agree upon. As the
years passed, there would be an ever widening rift between the two. Chain
explained his motive for working with penicillin:
I became interested - immediately - in Fleming's paper, not
because I hoped to discover a miraculous drug for the
treatment of bacterial infection which for some reason had
been overlooked, but because I thought it had great scientific
interest. In fact, if I had been working at that time in aimdirected scientific surroundings, say in the laboratory of a
pharmaceutical firm, it is my belief that I would never have
obtained the agreement of my bosses to proceed with my
project to work with penicillin.
I thought this would be a relevant passage to quote because in many
universities, this one in particular, there are those politicians who wish to
restrict apparently purely academic research because they see no value in
its pursue. They should probably read this passage by Chain.
It was also Chain who began immediate work with penicillin by extracting
what he believed to purified penicillin. After a great deal of effort,
enough was extracted for experimentation to begin. Tests began with
laboratory animals. Initially only two mice were tested. Chain was anxious
to begin, but being a biochemist by training, he was not qualified to inject

laboratory animals. Rather than Florey, Chain asked another colleague,

J.M. Barnes, to inject the two mice. The two mice were injected and were
unharmed. Chain explained Barnes involvement was due to the absence of
Florey who was away from Oxford and explained that he had approached
Florey four times in several weeks about doing the injections. The last
time he requested his assistance, Florey turned to Mrs. Margaret
Jennings, who was in the lab, and said, pointing to Chain, "in one of my
weak moments I promised this man to test his fractions and here he
comes pestering me again." After what Chain described as "these
humiliating remarks in front of others not involved in the project, it
became clear to me that Florey was not really interested in penicillin"
This was the reason for asking Barnes rather than Florey to carry out the
inoculations. As another digression, following this successful inoculation,
Florey was not at all pleased with this test being done in his absence.
Chain maintained that Florey was not interested in participating until
after he had carried out this test successfully.
Following the successful injection of two white mice, fifty white mice
were inoculated with deadly Streptococcus germs, followed by injection
of penicillin in half the mice. The second group, did not receive penicillin.
All of the latter group died the next day while the former mice all
recovered. Florey now thought they were ready to test penicillin on human
subjects. However, it would be difficult to produce enough penicillin, For
initial tests, by Florey, in 1940, on human subjects, it had required two
professors, five graduates and ten assistants working almost every day of
the week for several months to produce enough penicillin to treat six
patients.
The first real test for the Florey lab came when Albert Alexander, a 48
year old London policeman had nicked himself while shaving. It was only a
minor cut and was ignored for several days, but this would have a major
impact on medical history. After a time, Alexander's face became swollen

and infected and developed a temperature that rose to 105F. The minor
cut had developed septicemia. When he was rushed to Radcliffe Hospital,
sulfa drugs were administered without success. When the doctors
believed that the policeman only had several more hours of life remaining,
Florey and Chain requested that they be given permission to treat this
patient with their "purified" penicillin. This would be their first human
subject to which penicillin was applied. Florey and Chain took charge of
the case personally and injections of penicillin were begun. After five
days, the impossible had happened, the patient was recovering.
Unfortunately, because penicillin was in such short supply, that small
amounts had to be extracted from the patient's urine. after five days,
the supply of penicillin that was produced for experimentation had been
used up. There was no more penicillin available, and the policeman had a
relapse and died five days later. However, the experiment did
demonstrate that penicillin was not harmful to humans and was effective.
Penicillin had temporarily halted the infection, but in the end it had failed
in its first test. Unfortunately, because the patient had died, there were
a number of macabre jokes that came about, one that is well known, even
to this day: "the treatment was a success, but the patient died."
The next two treatments, however, would be successful. The first was a
15 year old boy who had a form of blood poisoning from a badly inflamed
hip joint and the second was a 48 year old laborer who had a large
carbuncle on his back. In both cases, the penicillin treatment was a
success. Encouraged with this success, they next tested penicillin on a 4
year old boy who was ill from an infection, following measles. Sulpha drugs
were used unsuccessfully. The boy, by this time, had developed an
infection in one eye socket which spread to the base of the skull and was
semi-comatose and it was believed that he had contracted meningitis by
this time. Penicillin was not tried until the infection had already
progressed to this advanced state. On the third day of penicillin
treatment, the swelling in the boy's eye had retreated and he was

emerging from the coma. The infection had been defeated and the
penicillin injections stopped in order to preserve the supply of penicillin.
Then suddenly the boy had a sudden convulsion and he died four days
later. Although the boy died, the autopsy revealed that he had died of a
burst blood vessel in his brain, and that the infection had been cured by
the penicillin injections.
During this period Fleming was not idle. Fleming contacted Florey to
request penicillin for the treatment of an old friend Harry Lambert, who
was in critical condition, suffering from meningitis. Sulfa drugs had
already been used and without success. Florey thought the procedure
planned by Fleming a risky one, but did supply him with the penicillin.
Fleming injected the penicillin into his friend's spine and the treatment
proved completely successful, with his friend making a miraculous
recovery. However, Florey had, at the same time, injected penicillin into
the spines of artificially infected animals, with the result that they had
all died! Luckily, Fleming did not learn about Florey's result until after he
had injected Lambert. He probably would have hesitated to give the
injection resulting in the death of his friend. Although the relationship
between Florey and Fleming seemed to be an amiable one. It would later
turn sour.
By this time, it was now 1941, it was now acknowledged that penicillin was
indeed a worthwhile drug and could save thousands of lives.
Unfortunately, the process that Florey and Chain developed, for
extracting penicillin, was producing only one part penicillin per million part
of culture medium. Thus, their penicillin was not purified as they thought
it had been. The amount was small indeed, and scientist could be heard
saying, "you could get more gold out of ordinary seawater than penicillin
out of the mold."

With World War II raging in Europe and the German bombardment of

England, all of England's resources and efforts had to be placed into the
war effort. Florey and Chain would be unable to utilize any factory
resources in England to experiment further with producing penicillin in
adequate amounts to be useful for large scale usage. Also, it was feared
that even if a factory could be utilized, it was feared that all their
efforts cold possibly be lost if the factory was bombed.
However, during the summer of 1941, Florey had negotiated an agreement
with the Rockefeller Foundation, which had been funding his research, to
fly he and one of his assistant to the United States to continue his work
with penicillin. The United States, who was at that time a neutral country,
would enter into World War II in another few months. This gave added
incentive to the penicillin project which became declared a war project
and was given top priority.
Despite the efforts and resources that were being given to producing
large quantities of penicillin, it soon became obvious that Fleming's
original culture would not be able to produce enough penicillin regardless
of the environment in which the fungus was grown. By 1942, there was
only enough penicillin produced to treat a few hundred people. There are
many species of Penicillium, and a search was started to find other
species that could be tested for penicillin production. Eventually, one was
found, on a moldy cantaloupe in a market in Peoria, Illinois. This species
would be identified as Penicillium chrysogeum, and would produce
approximately 200 times as much penicillin than P. notatum. This is the
species that is currently used to produce penicillin. However, even this
amount would be inadequate to produce the amount that would be
required.
Scientist then began to try to increase the amount of penicillin produced
by P. chrysogenum, by irradiating it with X-rays and UV rays in order to

induce mutations of this species. This eventually lead to a mutant that

produced 1000 times the amount of penicillin than Fleming's original
culture. In addition to the development of this mutant, a new means of
growing the mold was also perfected. Previously, penicillin was grown in
flask, the size of milk bottles, and hundreds of bottles of Penicillium

notatum were needed to produce enough penicillin for only a single person.
The new method involved growing the mold in large metal tanks, which
held 25,000 gallons of nutrient, were aerated so that the mold could grow
throughout the entire tank rather than on top. Aeration was the key to
growing it in such large tanks. Previously, this had not been tried because
it was known that the mold would only grow on the surface of the liquid
medium. Thus, utilization of a large tank, under such circumstances would
be highly inefficient in terms of cost, space and penicillin production.
With this new method, production quantity began to rise. In 1943, 29
pounds were produced, and with increases in the number of
pharmaceutical companies producing penicillin, there was a tremendous
increase. By the end of the war, enough penicillin was produced to treat
seven million patients/year.
With respect to the war, the use of penicillin was immediately apparent.
During World War I, death rate from pneumonia in the American Army
totaled 18%. In World War II, it fell to less than 1%.
One illness after another, that was tested, was cured by penicillin, which
was by this time dubbed a "wonder drug." In addition to pneumonia and
blood poisoning, the major causes of death, in hospitals, during the war,
strep throat, scarlet fever, diphtheria, syphilis, gonorrhea, meningitis,
tonsillitis, rheumatic fever, and many other diseases were successfully
treated with penicillin.
During this period of time when mass production of penicillin was being
perfected, Fleming had little to do with penicillin. Florey did update him

as to the progress being made and samples were often sent to him, but by
this time Fleming was no longer part of penicillin research. Although,
Fleming was the discoverer of penicillin, he was almost forgotten by the
time penicillin was being mass produced. However, he would be rescued
from oblivion, in 1943, when he was knighted along with Howard Florey,
and two years later was awarded the Nobel Prize in Physiology and
Medicine along with Florey and Chain.
So Fleming eventually did get credit for the discovery of penicillin, but it
was certainly not an overnight success. Also, Fleming had little to do with
penicillin while research was being done with regards to its application and
mass production. Thus, Fleming's role in penicillin was not that of a lone
researcher discovering a wonder drug, but rather one of many people that
contributed to the successful exploitation of penicillin in medicine.
Penicillin brought about the biggest search in medical history. It was
reasoned that if there was one antibiotic in nature, there must be many
more, and many more would later be found. However, few would be fungal
in origin. Most of the later antibiotic discovered would be derived from
bacteria, specifically Actinomycetes. Yet, without the discovery of
penicillin, all of these other antibiotics would possibly never been
discovered.
As a postscript, Fleming would, unfortunately, make a prediction that
would come true. That the use of penicillin would, in time, be of limited
value because bacteria would eventually recombine genetically to resist
the effects of penicillin. By as early as 1952, as much as three-fifths of
all staph infections were penicillin resistant. Various steps were taken so
as to continue the use of antibiotics. New antibiotics are constantly being
sought for this reason. Other approaches include using combination of
antibiotics and changing the chemical structure of antibiotics in the
laboratory so that all of slightly different properties. These attempts

have all been tried and have been successful, but unfortunately, the
bacteria are still recombining genetically to become resistant to these
new efforts.
Penicillin Terms
Alexander Fleming: Person often credited with the discovery of penicillin
and its properties. For this discovery, he was knighted in 1943 and was
one of three scientists to receive the 1945 Nobel Prize, in physiology and
medicine. The other scientists were Howard Florey and Ernst Chain.
Antibiotic: A chemical substance that is used in the treatment of
bacterial infectious diseases and has the ability to either kill or inhibit
the growth of having the capacity in dilute solutions to either kill or
inhibit the growth of certain harmful microorganisms.
Chain, Ernst: One of the main scientists who worked with Florey in the
research with penicillin, and one of the three scientist to share the Nobel
Prize, in 1945, for their work on penicillin, in physiology and medicine.
The other two scientists were Howard Florey and Alexander Fleming.
Florey, Howard: In1938, directed his efforts to doing research with
penicillin, at Oxford. His research program eventually was the one that
led to a method in the purification and mass production of penicillin. This
led to a Knighthood, in 1943, and the Nobel Prize, in 1945, in physiology
and medicine. The later was shared with Ernst Chain and Alexander
Fleming.
Germ Theory of Disease: Concept put forth by Pasteur that each disease
is caused by a specific type of microorganisms.
Magic bullet: A theoretical substance that could kill disease, causing
organisms, without harming the infected organism.

Paine, Cecil: The first scientist to demonstrate the potential usefulness

of penicillin, in human patients. Paine used a crude extracts of penicillin
from a culture of Penicillium notatum, and was able to save a miners
injured eye that had become badly infected and would ordinarily have
been removed. Also, treated a case of hereditary gonorrhea that
prevented blindness in the child. Normally, children with such diseases
would have become blind. Although it was disputed by Florey, Paine told
Florey of his success with using penicillin on human patients, which
probably stimulated his interest in working with penicillin.
Penicillin: First antibiotic isolated and used in treatment of bacterial
diseases and infections.
Septicemia: Also called "blood poisoning", it is caused by virulent
microorganisms that have invaded the bloodstream, usually through a local
infection.
Sulfa Drugs: A class of synthetic chemical substances developed during
the mid 1930s for treatment of certain of bacterial infections.
Questions to Think About
1

Why was knowledge of the Germ Theory necessary before thought

could be given to search for the "Magic Bullet"?

Although Alexander Fleming is usually given credit for the discovery

of penicillin, why is it unfair to single him out as the only person to
receive credit for this discovery?

Fleming was not the first to observe the inhibition of bacterial

growth by fungi. Who were some other people that made the same
observation? What do you suppose was the reason that some of
them did not follow up on this observation?

In World War I and World War II, many soldiers wounded in battle
died, not as the result of their wounds, but for another reason.
What was the main reason that these soldiers were dying after
their wounds had been successfully treated?

Fleming was unable to further his research in penicillin after making

a crude extract of penicillin, from P. notatum. Why was Florey
successful in continuing Fleming's work on penicillin while Fleming
could not?

What was a possible reason as to why Florey suddenly switched the

direction of research, in his lab, from lysozymes to penicillin, in
1938.

Although P. notatum was the species in which Fleming first

discovered penicillin and it was that species (in fact the same
culture that Fleming isolated) that Florey and his lab continued
their work with penicillin, his lab eventually began using another
species, P. chrysogeum, to produce penicillin. Why was there a
change in species?

Although penicillin and a number of other antibiotics that followed

are presently used to treat various bacterial diseases and
infections, with each passing day, these antibiotics are becoming
less and less effective. Why?

History of penicillin
Penicillin has been used throughout history to fight disease, but it was not until 1928
that it was officially discovered. In 1928, Alexander Fleming was conducting a
laboratory experiment, and incidentally ran into the fact that the Penicillium fungus
had strong antibacterial properties. A list of significant events leading up to Flemings
discovery follows:

- 1640 John Parkington recommended using mold for treatment in his book on
pharmacology - 1870 Sir John Scott Burdon-Sanderson observed that culture fluid
covered with mould did not produce bacteria - 1871 Joseph Lister experimented with
the antibacterial action on human tissue on what he called Penicillium glaucium 1875 John Tyndall explained antibacterial action of the Penicillium fungus to the
Royal Society - 1877 Louis Pasteur postulated that bacteria could kill other bacteria
(anthrax bacilli) - 1897 Ernest Duchesne healed infected guinea pigs from typhoid
using mould (Penicillium glaucium) - 1928 Sir Alexander Fleming discovered
enzyme lysozyme and the antibiotic substance penicillin from the fungus Penicillium
notatum
While Flemings discovery was a major step towards antibiotics, it was not until the
1940s that penicillin could be used as a medicine. [2] Howard Florey and Ernst
Chain were able to develop upon Flemings findings and ultimately isolate the active
ingredient, penicillin, and create pills used to fight bacterial infections.[2]

How It Works
Penicillin works by preventing cells from dividing. It does not allow them to
synthesize cell wall, and thus when the cells attempt to duplicate, they rupture and
end up killing themselves. Because penicillin has such a strong focus on a bacterias
cell wall, it is far more effective on Gram-positive organisms. When a Gram-positive
organism attempts to duplicate itself, it must create more cell wall and split off, the
penicillin causes there not to be any new cell wall, and instead of duplicating, the cell
will simply rupture, effectively killing it.[3]

Diseases Cured with Penicillin

Penicillin antibiotics are most effective against gram-positive bacteria, e.g. the
genera bacillus, clostridium, streptococcus, and staphylococcus). There are many
different bacterial infections, diseases, and conditions that have been combated with
the help of Penicillin. Here is a brief list of examples: Chlamydia Penicillin can be
prescribed to pregnant women in order to prevent the child from contracting the
disease as well.[4] Stomach Ulcers Penicillin was found to be effective in
controlling the effects of stomach ulcers.[4] Tooth Abscesses Penicillin kills the
bacteria responsible for causing these infections.[4] Step Throat and Scarlett Fever caused by the Group A Streptococcus bacterium streptococcus pyogenes, which
penicillin can kill. [5] [6] Staph Infections - can lead to amputation of affected limbs
and appendages if left untreated, or even death in cases in which the infection
enters the blood stream. Fortunately, Penicillin was found to be effective in killing
staphylococcus aureus, the bacterial cause of the infections.[7] Leptospirosis (Weils
Syndrome) Can cause kidney and liver failure if left untreated.[8] Lyme Disease

Penicillin can help prevent the disease from spreading throughout the body; which
could lead to further complications with the heart and nervous system in the later
stages of the disease.[9] Typhoid Fever can lead to kidney failure and eventually
death if left untreated. Before penicillin, was much more widespread in the US;
35,000 reported cases during the 1920s, about 400 cases reported annually in
modern times.[10] Gas Gangrene can prevent further spread of this condition
which is caused by a clostridium perfringens infection.[11] Necrotizing Fasciitis
(Flesh-eating disease) destroys soft tissues around infection site; will spread
without treatment.[12]
While Penicillin antibiotics were at first extremely killing the bacteria that cause these
diseases, the bacteria soon built up immunity to these antibiotics as they were
overprescribed. Due to this, many bacterial strains are now resistant to Penicillin,
and some of these diseases are no longer able to be treated using Penicillin
antibiotics. This development has led pharmaceutical companies to have to produce
new antibiotics that work in the same way as Penicillin antibiotics once did.

Developments from Penicillin

Although penicillin itself has a narrow spectrum of activity, virtually every antibiotic
discovered or developed post- penicillin owes its existence to it. Not only do the
penam sub-class of Beta- Lactam antibiotics stem directly from this first miracle
drug; but the discovery of penicillin also ignited the desire in scientists to find and/
or develop other antibiotics as well (the first of which was streptomycin, which
effectively cured Tuberculosis). The penam subclass, which is the most widely used
group of antibiotics, consists of penicillin and its derivatives- all of which end in
cillin and possess a central Beta- Lactam ring structure. The penams can be
broken up into two camps: Extended spectrum and Narrow spectrum antibiotics. The
extended spectrum penams include amoxicillin, ampicillin, and mezlocillin, among
others, and as their name suggest, these antibiotics work against very wide range of
bacteria. Some extended spectrum penams (called antipseudomonal penicillins)
even proved useful against Gram-negative bacterial cells, including pipericillin,
carbenicillin, and ticarcillin.
The Narrow spectrum antibiotics, on the other hand, act on a smaller, more specific
group of bacteria. Narrow spectrum drugs can be broken up into those that are BLactamase sensitive (e.g.- benzylpenicillin, azidocillin) and those that are BLactamase resistant. B- Lactamase is an enzyme that many bacteria started
producing as a means to resist the B- Lactam antibiotics. New antibiotics, the BLactamase resistant set, were then developed to combat the bacterias resistance to

penicillin and penicillin- like drugs. Antibiotics that are resistant to the B- Lactamase
enzyme, and thus can kill bacteria that the other penams cannot, include oxicillin,
flucloxicillin, and methicillin; however, only two years after methicillin hit the market,
strains of Staphylococcus aureus began exhibiting signs of resistance to it. Now
Methicillin- resistant Staphylococcus aureus (MRSA) has become one of the
infections most feared in hospitals and on athletic fields.

The Future of Penicillin

Bacteria are often able to become resistant to antibiotics quickly, which makes
staying ahead of bacteria when developing antibiotics very difficult. What causes this
is that when antibiotics are used such as penicillin, if there are any microbes left
behind, they begin to repopulate with resistances to the antibiotic. These resistances
easily come as microbes share their information about the antibiotics, which causes
a need to develop more antibiotics; however this takes a lot of time and money and
is not very profitable. Staphylococcus aureus is a gram-positive microbe that was
the first bacteria penicillin was used against. The infection combated penicillin by
acquiring an enzyme, B- Lactamse, that would snip the central ring of penicillin. By
doing this, the microbe became resistant to penicillin and now most of
Staphylococcus aureus cannot be fought with penicillin. Microbes can become
resistant to antibiotics in a couple ways. If an infection alters a protein that the
antibiotic uses to bind to in a spontaneous mutation, the antibiotic would not be able
to work. Infections can also acquire new instructions. They can pick up new DNA
that carries information allowing the bacteria to modify such as Staphylococcus
Aureus did in its ability to snip the central ring of penicillin. New DNA can be shared
between different species of bacteria and this sharing is often random however the
microbes that gain the DNA leading to resistance will survive. This new DNA can be
picked up from a dead cell, passed from cell to cell or transferred through viruses.
There are also ways for us to combat the growing resistances of microbes. By
limiting out own use of antibiotics, bacteria will have less exposure to the antibiotics.
This would cause it to be more difficult for them to develop resistances. To do this,
doctors must be educated to prescribe antibiotics only when they are actually
needed. When antibiotics are prescribed for viruses they are not effective and only
help microbes build up resistance to the microbes. Also patients should make sure
to take the full course of antibiotics. When stopping taking antibiotics early or just
when symptoms subside, there could still be a few infectious microbes left which
would re populate with greater resistance to that antibiotic. There is also a lot of use
of antibiotics on farm animals, which can cause a transfer of the animals
resistances into us. Even with the growing resistance there is a declining
development of new antibiotics for economic reasons. From 1983 to 1987 there
were 16 new antibiotics made and approved by the FDA, from 2003-2007 there
were 5, and since 2008 there have only been 2. This is because there are now far

fewer pharmaceutical companies investing in making new antibiotics because there

are not profitable ways to produce new antibiotics. Top selling drugs in 2009 where
Lipitor, Plavix, remicade, advair, Enbrel, avastin, ability, rituxan, humira, diovan,
crestor, and lovenox. None of these are antibiotics and all are used for chronic
conditions which is a much more profitable field for developing drugs. In order to
continue to develop new antibiotics there would need to be financial incentives for
the companies to develop new antibiotics and increases in funding. Because this is
unlikely there is not an expectation for many new antibiotics until there is a dire need
which would create a profitable demand in the economy and become beneficial for
companies to develop new antibiotics.

Maker of the Miracle Mould

The story of penicillin - the first antibiotic used successfully to treat people with serious
infectious diseases - begins with a bit of luck. Alexander Fleming, a British scientist,
noticed in 1928 that mould had prevented the growth of bacteria in his lab. But the main
plot of the story involves the rediscovery of penicillin 10 years later by an Australian
scientist born one hundred years ago this year. Howard Florey and his dedicated team's
systematic, detailed work transformed penicillin from an interesting observation into a
life saver.
Emma Burkervisc used to be the tea lady at the Australian National University's John
Curtin School of Medical Research (Howard Florey played a crucial role in the
establishment of the School and University later in his life). Emma's life was saved by
penicillin in a German refugee camp after World War II. Imagine how she felt many
years later, bumping into Florey in the corridors where she worked - the man who made
the supply of penicillin possible. Imagine how you'd feel if you met someone who saved
your life; and think of Florey and his team's impact on the world.
Breaking the Mould
Florey gathered a team of scientists at Oxford University in Britain in the 1930s, when
working together on scientific discoveries as a group was not at all common. Nowadays
scientists work together all the time, but Florey realised that science had reached a
point where a team of specialists was needed - the job was too big for one person.
His team commenced a careful investigation of the properties of anti-bacterial
substances that are produced by mould. One member of the team, Ernst Chain, found

an article about Alexander Fleming's work while flicking through a medical journal, and
this prompted them to begin looking at penicillin.
Individual members of the group concentrated their attention on areas in which they had
the most knowledge, but they often met to exchange ideas. Chain worked on purifying
penicillin with Edward Abraham. Norman Heatley improvised methods for extracting
penicillin using ether and bedpans (see 'penicillin production' below). A. D. Gardner and
Jena Orr-Ewing studied how penicillin reacted with other organisms. Howard Florey
looked with Margaret Jennings at the impact of penicillin on animals. Ethel Florey later
worked with her husband on clinical trials of penicillin.

In May, 1940 they performed one of the most important medical experiments in history.
The work was so urgent that they came in to begin the experiment on the weekend, and
on Saturday 25 May, Florey's team tested penicillin on eight mice injected with a lethal
dose of streptococci bacteria. Four of the mice were treated with penicillin, while four
were used as controls. By the next day, the treated mice had recovered and the
untreated mice were dead. In the early days of World War II, the lives of eight mice may
seem insignificant. But their rescue by penicillin led to the treatment of Allied soldiers as
early as D-Day, in June 1944, and probably influenced the outcome of the war.
The First Patient
The results were so exciting Florey knew that it was time to test the drug on humans.
The first patient in 1941 had been scratched by a rose thorn. Albert Alexander's whole
face, eyes and scalp had swollen. He had already had an eye removed and abscesses
drained; even his remaining eye had to be lanced to relieve the pain of the swelling. He
was given penicillin, and within a day he began to recover. But Florey's team didn't have
enough of the drug to see the patient through to a full recovery. Their efforts to recycle
the penicillin by extracting it from his urine failed, and he unfortunately had a re-lapse
and died. Because of the awful experience, the team then concentrated their efforts on
sick children, who did not require such large quantities.
In 1943 Florey travelled to North Africa to test the effects of penicillin on wounded
soldiers. His trials were seen as a miracle. Instead of amputating wounded limbs or
simply leave them to heal, he suggested soldiers' wounds be cleaned and sewn up, and
that the patients then be given penicillin. Thanks to Florey and his team, the drug was
available to treat Allied troops by the end of World War II. It has since revolutionised
medical science, saving millions of lives.
Before Penicillin

How many times have you accidentally pricked your finger on a rose thorn, or perhaps a
sewing needle? Nowadays, if the wound became infected you'd be cured almost
immediately. But before the second half of this century, you could have been in big
trouble. Infections were feared then as cancer is feared today. Your glands would swell
up and require lancing to release the pus. A surgeon might even have to amputate your
arm in an attempt to save your life. This was the nightmare of many infectious diseases
before Howard Florey developed penicillin.
Setting the Stage for Penicillin
Three thousand years before penicillin, moulds and fermented materials had been used
to cure various skin infections, although without an understanding of how they actually
worked. But it wasn't until the late 1800s that scientific studies of antibiotics began.
French chemist Louis Pasteur, after discovering that infectious diseases are spread by
bacteria, observed that mould inhibited the growth of anthrax (an infectious disease
spread from animals to humans). British surgeon Joseph Lister noted that samples of
urine contaminated with mould didn't allow bacteria to grow, but he was unable to
identify the substance in the mould. French medical student Ernest Duchesne
successfully tested a substance from mould that inhibited bacterial growth in animals,
but died at an early age in 1912, never seeing the world's acceptance and use of his
important discovery.
After World War I, Alexander Fleming was conducting an experiment with bacteria when
a tear fell from his eye into a culture plate. He later noticed that a substance in his tear
(which he named lysozyme) killed the bacteria, but was harmless to the body's white
blood cells. Years later, Fleming was doing research on the flu when a similar
coincidence occurred. While he was on holidays, a bit of mould had fallen into a
discarded culture plate containing bacteria, forming a clear patch. When he returned he
recognised this pattern from his previous experience with lysozyme. He concluded that
the mould was producing an antibiotic substance and named the antibiotic penicillin,
after the Penicillium mould that produced it.
His discovery was an amazing piece of luck. If Fleming hadn't left a petri dish of bacteria
on his bench when he went on holidays; if he had properly disinfected the dish; if the
weather had been different from the ideal conditions for bacteria and mould growth in
the laboratory; and especially if Fleming hadn't the experience to recognise the
importance of the observation, penicillin may not have been discovered as an antibiotic.
But Fleming couldn't extract the bacteria-killing substance, so he couldn't try it as a
treatment for general infections. He moved on to other research - leaving Howard Florey
and his team to pave the way for penicillin's use as a lifesaver more than a decade later.
Fabulous Fungus

Penicillin was the first naturally occurring antibiotic discovered (Prontosil, the first
chemical used to cure certain infectious diseases, had been discovered in 1933 but had
serious side effects). There are now more than 60 antibiotics, which are substances that
fight bacteria, fungi and other microbes harmful to humans - the word means against
(anti) life (bio).
An antibiotic is a drug produced by microbes. Penicillin is obtained in a number of forms
from Penicillium moulds. Penicillin G is the most widely used form, and is the one that
killed bacteria during Duchesne's work in 1896, Fleming's work in 1928 and Florey's
work in 1939.
Bacteria reproduce by dividing to produce two new cells. They enlarge to about twice
their size before the DNA chromosome is copied. The two new chromosomes move
apart and a cell wall forms between them. But if penicillin is around the new cell wall
won't be able to form. It doesn't harm old bacterial cell walls, it just stops new ones
forming (see Balloon bacteria). This means the bacteria can't reproduce, so the disease
can't spread.
Natural penicillin is administered by injection, because if it's swallowed stomach acids
destroy the drug before it reaches the bloodstream. One shot of penicillin these days is
more than the entire amount used by Florey's team in all its clinical trials! About one in
10 people is allergic to penicillin, showing symptoms ranging from minor rashes to
serious breathing difficulties. If you're allergic to penicillin, there are now other
antibiotics that can be taken as a substitute.
Penicillin Production
Florey's team worked under difficult circumstances with a lack of funding and
equipment, but ensured penicillin production grew from the manufacture of a scarce and
very impure brown powder to the commercial production of a purified and powerful
antibiotic.
At first penicillin was made using old dairy equipment. Hospital bedpans were used to
grow mould. Liquid containing penicillin was drained from beneath the growing mould
and filtered through parachute silk on bookshelves. But the team needed drug
companies to help it produce the large amounts required for test patients.
Companies in Britain were unable to help out on a large scale because of the war, so
Howard Florey and Norman Heatley took a dangerous flight to the United States in a
blacked-out plane across the Atlantic. The trip was against the wishes of Ernst Chain,
who wanted to first patent their ideas in Britain. This would have made the team very
rich indeed, but it was thought in Britain at the time that patenting medical discoveries
was unethical.

Florey explained his penicillin-making methods to people in the US, and there happened
to be a Department of Agriculture laboratory looking for a new use for a thick liquid that
was a by product from the corn-milling process. When this liquid was used, 10 times the
amount of penicillin was able to be produced than before. Mary Hunt, known as Mouldy
Mary for her enthusiasm in finding new sources of mould, then found mould growing in
cantaloupe (rockmelon) was twice as successful again at producing penicillin.
By late 1943, mass production of the drug had commenced - only four years after the
first mouse experiments and in spite of the war, a sign of Florey's persistence and
determination. By the end of the war, many laboratories were manufacturing the drug,
including the Merck, Squibb and Pfizer companies in the US and the Commonwealth
Serum Laboratories in Australia. In fact, Australia was the first country that made the
drug available for civilian use.
However, several strains of bacteria became resistant to penicillin after a few years,
through mutation of the cells. To overcome this problem, scientists in the 1950s made
artificial penicillin by chemically changing natural penicillin. Resistant bacteria multiply
when non-resistant bacteria die. Hospitals in Australia and around the world are now
seeing the arrival of antibiotic-resistant bacteria due to the overuse of antibiotics,
exposing the very young, very old and very sick people to infections and diseases.
So Who Was Howard Florey?
Florey's father owned a shoe business in Adelaide, having migrated from England for a
warmer climate due to his wife's poor health. When she died of tuberculosis, he married
Bertha and they had two daughters, followed by Howard.
Howard was brilliant at schoolwork and outstanding at sport. He was inspired by his
high school chemistry teacher to study medicine at the University of Adelaide. Here he
met Ethel, a fellow medical student. He won the South Australian Rhodes Scholarship
(a prize awarded for excellent leadership and determination in academia and sport) and
went to Oxford University at the end of 1921. Howard and Ethel wrote to each other for
a few years, before she moved to England to marry him in 1926. After an unhappy
marriage - partly due to her poor hearing and health, partly due to his intolerance - she
died in 1966. Howard married again in 1967 - Margaret Jennings, an important member
of the penicillin team - just one year before he died of a heart attack, aged 69.
His role as the leader of the team of scientists that discovered and developed penicillin
won him the Nobel Prize in 1945 with Fleming and Chain. He was knighted in 1944, and
was decorated around the world by the UK, France, Australia and the US for his
influence on the outcome of World War II. He was the first Australian elected to the
prestigious position of President of the Royal Society in 1960, where he was known as

'the Bushranger President'. He was made Baron Florey of Adelaide in 1965 and
accepted the Chancellorship of the Australian National University in 1965.
Brilliant But Humble
Florey was humble about his achievements, describing them in an interview in 1967 as
a "terrible amount of luck" that "involved many others." He said, "All we did was to do
some experiments and have the luck to hit on a substance with astonishing properties."
He was a quiet man, but sure of himself and his direction. He inspired those around him
with his scientific enthusiasm, his skill and his total honesty and lack of pretentiousness.
Florey was aware of the effect his drug had on population growth. He said in 1967, "I'm
now accused of being partly responsible for the population explosion... one of the most
devastating things that the world has got to face for the rest of this century." Perhaps to
counter this effect, contraception research was a lifelong interest and he was an active
advocate for population control.
He avoided the media when penicillin's success was discovered because he disliked interviews
and didn't want to create a huge demand for the scarce supply of the drug. Although a suburb of
Canberra is named after Howard Florey and his face appeared on the old $50 note, his dislike
of publicity may be why his vital role in the penicillin story is still largely unknown around the
world. Let's hope that on the anniversary of his birth, the true story becomes clear.