ADIGRAT UNIVERSITY

COLLEGE OF ENGINEERING AND TECHNOLOGY

DEPARTMENT OF CHEMICAL ENGINEERING
PROJECT ON: PENICILIIN PRODUCTION
COURSE TITLE: PROCESS INDUSTRY II
COURSE CODE: ChEg 3142
PREPARED BY NETWORK THREE
NAME
1. ETAY HAILU

IDNO
RET0457/06

SECTION

ONE

2. H/MARIAM ASGEDOM RET0748/06

ONE

3. KIROS G/MEDHIN

RET0937/06

ONE

4. LETU DESALEGN

RET0969/06

ONE

5. LICHIYA ALEM

RET0971/06

ONE

6. WASIE KEBRIE

RET1578/06

ONE

SUBMITTED TO:
INSTRUCTOR G/KIROS ARAYA
SUBMISSION DATE:
01, 10, 2008 E.C

ACKNOWLEDGMENT
First and for most we would like to say thanks for our instructor G/kirosAraya to give this
project and the formats of the project. Moreover, for all group members and for our advisor that
supports by giving idea and economical aid until the project is complete.

ABSTRACT

Penicillin antibiotics were among the first drugs to be effective against different diseases.
The discovery of penicillin and its medicinal uses was arguably the most important scientific
discovery of the 20th century. As new ways were found to ensure that more penicillin was being
produced and that the purification process was as effective and possible. Despite the expanding
number of penicillin resistant bacteria, penicillin are used to treat a wide range of infections
caused by certain susceptible bacteria Since then, the development of large scale production has
allowed penicillin to be used whenever needed to kill off bacteria and prevent serious infection.
The ability of some bacteria to now produce penicillinase to break down and render penicillin
completely useless has come about due to the wide scale use of the drug and has therefore
limited the effectiveness of penicillin as a clinical treatment.

II
LIST OF CONTENT
Contents

Page

Acknowledgment
Abstract

II

List of figures IV
Penicillin production 1
1. Introduction

1.1 Back ground

1.2 Statement problem

1.3 Objective 3
2. Literature review

2.1 General Structure of Penicillins 4

2.2 Specific Conditions for Penicillin Production
2.3 The Industrial Production of Penicillin
2.3.1

Stages of penicillin Production

2.4 Classification of Penicillins

3. Materials and methods

3.1. Materials 9
3.2. Methods 10
4. Enviromental impact

10

5. Result and discussion

11

5.1 Result

11

5.2 Discution

11

Conculision

12

Refference

13

III
List of figures page

5
5

Fig-1General structure of penicillin 4

Fig-2Effect of dilution rate on fermentor penicillin
Fig-3 seed culture

IV

PENCILLIN PRODUCTION
1. Introduction
Penicillins are a group of closely related antibiotics that kill bacteria.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 on set 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. There are several types of penicillins, each used to treat different kinds of
infections, such as skin infections, dental infections, ear infections, respiratory tract infections,
urinary tract infections, gonorrhea, and other infections caused by bacteria. These drugs will not
work for olds, flu, and other infections caused by viruses.

1.1 Background
You are all undoubtedly familiar with the story of penicillin. 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 moldPenicilliumglaucum.

Louis Pasteur and Jules Francois Joubert, in 1877, observed that cultures of the anthrax
bacilli, when contaminated with unidentified molds, became inhibited. But at the last
Alexander Fleming discovered penicillin by chance is a myth.

1.2Statement of the problem

The most common side effect of penicillin is diarrhea . Nausea , vomiting , and upset
stomach are also common. With some penicillins, particularly the broad spectrum
products, there is a risk of increased growth of organisms that are not affected by
penicillin.
This situation can lead to candidal infections of the mouth and vagina.Most side effects
of penicillin cannot be prevented. Amoxicillin has a lower incidence of diarrhea than
ampicillin and is the preferred drug in most cases.

1.3 Objective
Penicillins are useful against infections in many parts of the body, including the
mouth and throat, skin and soft tissue, tonsils, heart, lungs, and ears. However, since
many bacteria are resistant to penicillin, it is often wise to do a culture and sensitivity
test before using penicillins. In some cases, there are only a few types of bacteria that
are likely to be a problem, and so it is appropriate to use a penicillin without testing. For
example, dentists often prescribe penicillin to prevent infections after dental surgery.
To be able to identify useful products from microorganisms
To be able to identify the microorganisms used and the main stages in the
production of penicillin.
To be able to describe how Downstream processing is carried out to extract
and purify the endproduct of fermentation.

2.Literature Review
2.1General Structure of Penicillins

Fig-1 general structure of penicillin

Have -Lactam functional group, thus belong to the -Lactam antibiotic group.

They all have a basic ring-like structure (a -Lactam) derived from two amino acids
(valine and cysteine) via a tripeptide intermediate. The third amino acid of this tripeptide
is replaced by an acyl group(R).
The nature of this acyl group produces specific properties on different types of penicillin.

2.2 Specific Conditions for Penicillin Production

Most penicillins form filamentous broths. This means they can be
difficult to mix due to their high viscosity. Also the increasing viscosity of the broth can hinder
oxygen transfer.
A solution for the viscosity and the filamentous growth of penicillium species could be bubble
columns (air lift reactors) which would distribute the oxygen equally and also to
agitate the medium.
Penicillin is an aerobic organism; oxygen supply is critical: reactor
must have an efficient oxygen supply system
The optimum pH for penicillin growth is 6.5: maintain pH
Efficiently (pH controller and acid-base reservoir).
Strain Stability problems (mutations): careful strain maintenance is
required.
Biomass doubling is about 6h: provisions must be made.

2.3 The Industrial Production of Penicillin

This can be broadly classified into two processes.
1. Upstream Processing
- referring to processes before input to the fermenter and encompases any technology that leads
to the synthesis of a product.It includes the exploration, development and production.
2. Downstream Processing
- referring to processes done to purify the output of the fermenter until it reaches to the desired
product, such as extraction and purification of a product from fermentation.

2.3.1 Stages of penicillin Production

Medium for penicillin
1. The Penicilliumchrysogenumusually contain its carbon source which is found in corn steep
liquor and glucose.
2. A medium of corn steep liquor and glucose are added to the fermenter. Medium also consists
of salts such as MgSO4, K3PO4 and sodium nitrates. They provide the essential ions required
for the fungus metabolic activity.
Heat sterilization
3.Medium is sterilized at high heat and high pressure, usually through a holding tube or
sterilized together with the fermenter.
4. The pressurized steam is used and the medium is heated to 121C at 30 psi or twice the atm
Pressure.
Note: High temp and short time conditions are used to minimize degradation of certain
components of the media.
Fermentation
5. It is done in a fed-batch mode as glucose must not be added in high amounts at the beginning
of growth (which will result in low yield of penicillin production as excessive glucose inhibit
penicillin production).
6. The fermentation conditions for the Penicillium mold, usually requires temperatures at 2024C while pH conditions are kept at 6.5
7. The pressure in the bioreactor is much higher than the atmospheric pressure (1.02atm). This is
to prevent contamination from occurring as it prevents external contaminants from entering.
8. It is necessary to mix the culture evenly throughout the culture medium. Fungal cells are able

to handle rotation speed of around 200 rpm.

Fig-2 Effect of dilution rate on fermentor penicillin concentration, sugar

concentration &
Volumetric productivity of CSTR
Seed Culture
9. The seed culture is developed first in the lab by the addition of Penicilliumchrysogenum
spores into a liquid medium. When it has grown to the acceptable amount, it is inoculated into
the fermenter.

Fig-3 seed culture

10. The medium is constantly aerated and agitated. Carbon and nitrogen are added sparingly
alongside precursor molecules for penicillin fed-batch style. Typical parameters such as pH,
temperature, stirrer speed and dissolved oxygen concentration, are observed.
11. After about 40 hours, penicillin begins to be secreted by the fungus.

12. After about 7 days, growth is completed, the pH rises to 8.0 or above and penicillin
production ceases.
Removal of biomass
13. Filtration is carried out as bioseparation is required to remove the biomass from the culture
(removing the fungus and other impurities away from the medium, which contains the
penicillin).
14. A Rotary vacuum filter is commonly employed for filtration as it is able to run in continuous
mode in any large scale operations.
Addition of solvent
15. After filtration, phosphoric acid, a non-oxidising agent, is introduced,
to decrease pH from 8.0 to 6.5 so as to prevent loss of activity of penicillin.
16. Organic solvents such as amyl acetate /butyl acetate are added to dissolve the
penicillin present in the filtrate.
17. At this point, penicillin is present in the solution and any other solids will be considered
as waste (can be used as fertilizers and animal feed).
Centrifugal Extraction
18. Centrifugation is done to separate the solid waste from the liquid component which contains
the penicillin.
19. Usually a disk centrifuge is used at this point.
20. The supernatant will then be transferred further in the downstream process to continue
with extraction
Extraction.
21. A series of extraction processes are carried upon the dissolved penicillin, to obtain a better
purity of the penicillin product
22. The acetate solution is first mixed with a phosphate buffer, followed by a chloroform
solution, and mixed again with a phosphate buffer and finally in an ether solution.
23. Penicillin is present in high concentration in the ether solution and it will be mixed with a
solution of sodium bicarbonate to obtain the penicillin-sodium salt, which allow penicillin to
be stored in a stable powder form at rtp.
24. The penicillin-sodium salt is obtained from the liquid material by basket centrifugation, in
which solids are easily removed.

 Fluid bed drying

25. Drying is necessary to remove any remaining moisture present in the powdered
Penicillin salt.
26. In fluid bed drying, hot gas is pumped from the base of the chamber containing the
Powdered salt inside a vacuum chamber.
27. Moisture is removed this way, and this result in a much drier form of penicillin.
Storage
28. Penicillin is stored in containers and kept in a dried environment.
29. The resulting penicillin (called Penicillin G) can be chemically and enzymatically modified
to make a variety of penicillins with slightly different properties.
30. These can be semi-synthetic penicillins, such as; Penicillin V, Penicillin O, ampicillin and
amoxycillin.

2.4 Classification of Penicillins

1. Natural Penicillins:
The natural penicillins are based on the original penicillin-G structure.
They are effective against both gram-positive strains and gram negative bacteria.
Examples of Natural Penicillins are Penicillin G, Procaine, Penicillin O, Penicillin V and
Benzathine.
2. Penicillinase-Resistant Penicillins:
The Penicillinase-resistant penicillins have a narrower spectrum of activity in contrast to the
naturalpenicillins.
Their antimicrobial efficacy is targeted on penicillinase-producing strains of gram-positive
cocci, such as Staphylococcal species and these drugs are also known as anti-staphylococcal
penicillins.
Examples of Penicillinase-Resistant Penicillins are Dicloxacillin, Methicillin, Nafcillin,
Oxacillinand Cloxacillin.

3. Amino penicillins:
Active against gram-negative bacteria (such as E. coli).Aminopenicillins are acid-resistantcan be administered orally.
Orally administered amoxicillin and ampicillin are used primarily to treat mild infections
such as otitis media, sinusitis, bronchitis, urinary tract infections and bacterial diarrhoea.
Examples of Aminopenicillins are Amoxicillin, Ampicillin and Bacampicillin.
4. Extended Spectrum Penicillins:
These agents have similar spectrums of activity as compared to the aminopenicillins but it has
an additional activity, which is against several gram negative organisms in the family
Enterobacteriaceaeand some strains of Pseudomonas aeruginosa.

3. MATERIALS AND METHODS

3.1. Materials
pH 6.5
Temperature 20-24 C
Oxygen
Nitrogen: corn steep liquor 8.5 %
Glucose 1%
80% ethanol
phenylacetic acid
Probenecid
Lactose 1%
Calcium Carbonate 1%
Sodium hydrogen phosphate 0.4%
Antifoaming agent: vegetable oil

3.2 METHODS
In penicillin production there are two methods ;

1.Primary metabolism
is the metabolism of energy production for the cell and for its own biosynthesis. In aerobic
organisms (such as Penicilliumchrysogenum) it involves the conversion of sugars suchas glucose
to pyruvic acid and the production of energy.

2.Secondary metabolism:
regards the production of metabolites tha tare not used in energy production for example
penicillin from Penicilliumchrysogenum. The metabolite is being utilized as a defence
mechanism against other microorganisms in the environment. Penicilliumchrysogenumcan kill
off the competition to allow itself to propagate efficiently.
4. Enviromental Impact of Penicillin Production
There are five important impacts of the discovery of penicillin on society. One impact is now
bacterial infections are not troublesome . Penicillin forever altered the treatment of bacterial
infections and was recognized as the most life-saving drug in the world . A second impact is that
penicillin also has use in bacteriological media because it inhibits unwanted microbes in bacterial
cultures . A third impact of the discovery of penicillin is that this discovery triggered the Age of
Antibiotics, further research on antibiotics by scientists around the world, and advancement of
medicine. For instance, penicillin brought about the greatest search in medical history . People
began to think if there is one antibiotic, then there must be many more; and many more would be
found . Antibiotics are not only produced by certain fungi but are also derived from bacteria
particularly Actinomycetes . Nonetheless, without discovering penicillin, many other antibiotics
would probably never have been discovered. A fourth impact is a man-made negative impact.
Misuse or overuse of penicillin results in microbial resistance; the germ has taught itself to
outsmart its killer. With the convenience of penicillin, patients and doctors may have become too
quick to rely on a pill, disallowing the human body to heal itself naturally or alternatively. Taken

10

to an apocalyptic extreme, this could mean eventually all antibiotics lose their effectiveness and
humans begin to die from common infections.
5. RESULTS

AND DISCUSSION

5.1 RESULTS
There were major results from the research of penicillin. First, a specific type of
Penicilliumproduced a powerful antibacterial substance in culture . The antibacterial power of
the culture reached its maximum when incubated at a temperature of 20C (68F) . the
antibacterial power diminished until it had almost disappeared . Second, the best medium for the
production of the antibacterial substance was nutrient broth. Third, the active agent was filterable
and the name penicillin was given to filtrates of broth cultures and mold . Fourth, the active
agent was not destroyed by boiling for five minutes.
5.2 DISCUSSION
Penicillium species produce a very powerful antibacterial substance, which
affects the growth of a variety of bacteria . The least sensitive bacteria are
the Gram-negative bacilli while the most susceptible are the pyogenic cocci .
The inhibition is almost specific to microbes . The inhibitory substances are
hardly ever strong enough to withstand a slight dilution with fresh nutrient
mater. So, penicillin is not inhibitory to the original Penicillium used in its
preparation . Hence, penicillin can withstand additional penicillin, which is
important because it needs to be kept to a certain concentration in the body
in order to attain complete effectiveness.

11

Conclusion
Generally,Penicillins are a group of closely related antibiotics that kill bacteria.The various
antibiotics are used to treat a number of what are now common diseases and to prevent the on set
of infections when our skin, our first barrier to fight off disease, is somehow broken through a
simple cut or a more serious wound.

12

Reference
Hare,T; White,L/penicillin production
http://penicillin.wikispaces.com/General+bioprocess+flow
http://penicillin.wikispaces.com/Various+types+of+Penicillin
http://microbiologyprocesses.blogspot.com/penicillin-production.html
http//www.google.com

When life gives you molds,make

penicillin!

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