PENICILL

IN
PRAKSH DHAKAL

Public Health Microbiology

Tribhuvan University

INTRODUCTION :
First described by Fleming in 1929.
Isolated by Florey and Chain in 1940 from
Penicillium notatum.
The first clinical application of penicillin was in
1941.
Penicillins are produced by Penicillium and
Aspergillus species.
Penicillins belongs to the group of antibiotics
called -lactams which also include
cephalosporin,carbapenems,monbactams.
They are labile in acid and may be inactivated by
splitting the -lactam ring with -lactamases.

Currently the group of penicllins includes more than

20 antibiotics which are divided into several
categories.
1. Natural Penicillins :
( Penicllin G, Procaine, Penicillin G, Penicillin V,
Benzathine) The natural penicillins were the first
antibiotics introduced in clinical practice. Natural
antibiotics are active against gram-positive bacteria
such as Streptococcus spp, Staphylococcus spp.,
Bacillus spp., in less degree against Enterococcus spp.
L.monocytogenes, E.rhusiopathiae and corynebacteria
(including C.diphtheriae), the most of anaerobic
bacteria (Peptostreptococcus spp., Clostridium spp.) are
highly sensitive to natural penicillins. Natural
penicillins are highly effective against spirochetes
(Treponema, Borrelia, Leptospira).

2 . Penicillinase-Resistant Penicillins :
(Cloxacillin, Dicloxacillin, Methicillin, Nafcillin,
Oxacillin). Methicillin was the first member of
this group, followed by oxacillin, nafcillin,
cloxacillin and dicloxacillin. Currently Oxacillin
is the most widespread among penicillinaseresistant penicillins and is actively used for the
treatment of infections caused by staphylococci
including PRSA.

3.Aminopenicillins(Ampicillin, Amoxicillin,
Bacampicillin). Aminopenicillins were the first
penicillins discovered to be active against gramnegative bacteria (such as E. coli and H.
influenzae). Aminopenicillins are acid-resistant
and that is why are administered orally.
Aminopenicillins possess a broader spectrum of
antibacterial activity against some strains such
as E.coli, Shigella spp., Salmonella spp. and
P.mirabilis, which are characterized by low levels
of production of chromosomal -lactamase.
Aminopenicillins are susceptible to hydrolysis by
-lactamses and that is why are usually come in
combination with -lactamse inhibitors.

4.Extended Spectrum Penicillins(sometimes

called anti-pseudomonal penicillins). Extended
spectrum penicillins include both alphacarboxypenicillins (carbenicillin and ticarcillin) and
acylaminopenicillins (piperacillin, azlocillin, and
mezlocillin). These antibiotics possess similar
spectrum of antibacterial activity as
aminopenicillins but with additional bactericidal
activity against the majority of gram negative
microorganisms such as Pseudomonas aeruginosa.
Along with aminopenicillins, extended spectrum
penicillins are inactivated by beta-lactamase. These
drugs can be used in monotherapy of infectious
diseases or in combination with aminoglycosides.

Penicillins indication
The indications for penicillins vary depending on penicillins group.
Natural penicillins are used in:
Infections caused by S.pyogenes, and their consequences:
tonsilopharyngitis
scarlet fever
erysipelas
year-round rheumatism prevention.
Infections caused by S.pneumoniae:
community-acquired pneumonia
meningitis
sepsis
Infections caused by other streptococci:
Infective endocarditis (in combination with gentamicin or
streptomycin);
Meningococcal infections (meningitis)
Syphilis
Leptospirosis
Tick-borne borreliosis (Lyme disease)
Gangrenous emphysema
Actinomycosis

Penicillinase-Resistant Penicillins and namely

oxacillin is used in:

Staphylococcal infections of different localizations

and susceptible to oxacillin
Infections of skin, soft tissue, bone and joints
Pneumonia
Infectious endocarditis
Meningitis
Sepsis

Aminopenicillins are indicated for the treatment of:

Infections of upper and lower respiratory system
including sinusitis, aggravation of chronic bronchitis,
pneumonia
Urinary tract infections: cystitis, pyelonephritis
Meningitis caused by H.influenzae and L.monocytogenes
Endocarditis
Intestinal infections caused by Shigella and Salmonella
(ampicillin)
Eradication of H.pylori in patients with stomach and
duodenal ulcers
Prophylaxisof endocarditis
Intraabdominal infections
Skin and soft tissue infections

Carboxypenicillinsare usually indicated for the

treatment of:
Nosocomial infections caused by sensitive strains
P.aeruginosa. Carboxypenicillins must be used only in
combination with other antibacterial preparations
(aminoglycosides, fluorquinolones). Ticarcillin in
combination with clavulonate is indicated in:
Lower respiratory tract infections
Urinary tract infections
Intraabdominal infections
Sepsis
Infections of skin, soft tissue, bones and joints

ACTION MECHANISM :

All -lactam antibiotics exert bactericidal action

which is associated with inhibition of bacterial
cell wall production. Their target is penicillindepending bacterial proteins which fulfill a
function of enzymes on the final stage of
peptidoglycan synthesis-biopolymer and the main
component of bacterial cell wall. Suppressing of
peptidoglycan synthesis leads to consecutive
bacterial lysis. Some of penicillin antibiotic exert
bacteriostatic effect in low doses which may or
may not control the infection

CHEMICAL STRUCTURE :

The basic structure of penicillin is 6aminopenicillanic acid(6-APA), consisting of a

thiazolidine ring with a condensed B-lactam ring.

BIOSYNTHESIS AND REGULATION :

B-lactam thiazolidine ring of penicillin is
constructed from L-cysteine and L-valine.
Biosynethesis occurs in a non ribosomal process
by means of a dipeptide composed of L-alphaaminoadipic acid ( L-alpha AAA) and L-cysteine
or a breakdown product of cystathionine.
Then subsequently , L valine is connected via an
epimerization reaction resulting in the formation
of the tripeptide delta (L alpha-aminoadipyl)cysteniyl D-valine.

The first product of the cyclization of the

tripeptide which can be isolated is isopenicillin
N, but the biochemical reactions leading to this
intermediate are not understood.
Benzylpenicillin is produced in the exchange of Lalpha-AAA with activated phenylacetic acid.
Penicllin biosynthesis is affected by phosphate
concentration and also shows a distinct catabolite
repression by glucose, in addition to regulation by
concentration of ammonium ion, the latter by an
unexpalined mechanism.

STRAIN DEVELOPMENT :
The pencillin production of Flemings isolate was
about 2 international units/ml ; todays process
yield a penicillin titer of about 85,000 units/ml .
This is an increase from 0.0012 g/l to about 50 g/l .

PRODUCTION METHOD :
Penicllin G and V are produced using submerged
process in 40,000 to 200,000 liter fermenters
Due to difficulties with O supply ,larger tanks
2
cannot be employed.
It is a aerobic process with a volumetric oxygen
absorption rate of 0.4-0.5 mM/l-min.
The inoculum is started using lyophilized spores.
Spore concentration ( optimal 5x103/ml ) and
pellet formation are crucial for the subsqent
yield.
For optimal penicillin formation ,pellets must be
grow not as compact balls , but in loose form.

The growth phase is of 40 hrs duration cwith a

doubling time of 6 hrs, during which time the
greatest part of the cell mass is formed.
Oxygen supply should be maintained.
After growth phase penicillin production phase
occurs. In this phase growth is sharply reduced,
By feeding with various culture medium
components , the production phase can be
extended to 120-180 hrs.
The medium varies according to strain used and
usually consists of corn steep liquor ( 4-5 % by
dry wt) which may be replaced by other nitrogen
sources, in addition soy meal, yeast extract or
whey; a carbon source and buffers.

PH is kept constant at 6.5.

Phenyl acetic acid or pehnoxyacetic acid is fed
continuously as precursor.
About 65 % of the metabolized carbon source is
used for maintenance energy, 25% for growth
and only 10 % for penicillin production.
Yield increase of 25 % is reported by adding
glucose and acetic acid.
Pencillin is excreted into the medium and less
than 1% remains mycelium bound.

After separation of mycelium, product recovery is

accomplished by means of a two stage continuous
countercurrent extraction of the fermenter broth
with amyl or butyl acetate at 0-3Oc and PH 2.33.0 . The yield is around 90 %.

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