MODE OF ACTION AND AND INDUSTRIAL PRODUCTION OF SOME ANIBIOTICS

1. INDUSTRIAL PRODUCTION OF
ANTIBIOTICS
(Penicillin, Streptomycin, Tetracycline, Griseofulvin)
MAULSHREE
18MSCBT002
M.Sc. BIOTECHNOLOGY
3RD SEMESTER
MCE-JIBB.

2. CONTENT
• Introduction
• History
• Types of antibiotic
• Mechanism of action
• Penicillin
• Streptomycin
• Tetracycline
• Griseofulvin

3. INTRODUCTION
• Antibiotic: Any substance that can destroy or inhibit the growth of
bacteria and similar microorganisms
• Antibiotics are the secondary metabolites of microorganisms.
• Microbial production of antibiotics by secondary metabolism is one of the
key areas in the field of applied microbiology
• Antibiotics are produced industrially by a process of fermentation, where
the source microorganism is grown in large containers (100,000 –
150,000 liters or more) containing a liquid growth medium.
• During processing, the antibiotic must be extracted and purified to a
crystalline product.

4. HISTORY OF ANTIBIOTIC DEVELOPMENT
• 1928: Alexander Fleming
noted mould of the genus
penicillium contaminating
one of his cultures
preventing the growth of
bacteria.
• 1935: Domagk,
sulphonamide – synthetic
dye
• 1941: clinical trials of
penicillin- Florey and Chain

5. TYPES OF ANTIBIOTICS
• Bactericidal antibiotics kill bacteria directly,
• Bacteriostatic antibiotics stop bacteria from growing.
• Another important thing to remember about antibiotics is that they don't
work against all types of bacteria.
• Narrow-spectrum antibiotics are only effective against a narrow range of
bacteria,
• Broad-spectrum antibiotics are effective against a broad range of bacteria

6. MECHANISM OF ACTION
• Antibiotics act by disrupting various molecular targets within bacteria
and cell surface, preventing growth or initiating killing.
• 3 broad mechanisms:
• • Disrupt bacterial cell envelope
• • Block production of new proteins
• • Inhibit DNA replication

8. PENICILLIN
• Penicillin (PCN or pen) is a group of antibiotics which include penicillin G
(intravenous use), penicillin V (use by mouth), procaine penicillin, and
benzathine penicillin (intramuscular use).
• Penicillin was discovered in 1928 by Scottish scientist Alexander Fleming.
• • People began using it to treat infections in 1942.
• • Penicillin antibiotics were among the first medications to be effective against
many bacterial infections caused by staphylococci and streptococci.
• • They are still widely used today, though many types of bacteria have developed
resistance following extensive use.
• All penicillin are beta-lactam antibiotics.

9. • 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.
• Medium for penicillin
• The Penicillium chrysogenum usually contain its carbon source, which is
found in corn steep liquor and glucose.
• 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.

10. • Medium is sterilized at high heat and high pressure, usually through a
holding tube or sterilized together with the fermenter.
FERMENTATION
• It is done in a fed-batch mode as glucose must not be added in high
amounts at the beginning of growth The fermentation conditions for the
Penicillium mold, usually requires temperatures at 20-24°C while pH
conditions are kept at 6.5 The pressure in the bioreactor is much higher
than the atmospheric pressure (1.02atm). It is necessary to mix the
culture evenly throughout the culture medium. Fungal cells are able to
handle rotation speed of around 200 rpm.
• The seed culture is developed first in the lab by the addition of Penicillium
chrysogenum spores into a liquid medium. When it has grown to the
acceptable amount, it is inoculated into the fermenter.
• After about 40 hours, penicillin begins to be secreted by the fungus.
• After about 7 days, growth is completed, the pH rises to 8.0 or above and
penicillin production ceases

11. • Organic solvents such as amyl acetate /butyl acetate are added to dissolve
the Penicillin present in the filtrate
• Centrifugation is done to separate the solid waste from the liquid
component which contains the penicillin.
• Usually a disk centrifuge is used at this point. The supernatant will then
be transferred further in the downstream process to continue with
extraction.
• A series of extraction processes are carried upon the dissolved penicillin,
to obtain a better purity of the penicillin product.
• The penicillin-sodium salt is obtained from the liquid material by basket
centrifugation,
• Drying is necessary. Penicillin is stored in containers and kept in a dried
environment

12. STREPTOMYCIN
• Streptomycin is an antibiotic used to treat a number of bacterial infections.
• This includes tuberculosis, Mycobacterium avium complex, endocarditis,
brucellosis, Burkholderia infection, plague, tularemia, and rat bite fever.
• For active tuberculosis it is often given together with isoniazid, rifampicin, and
pyrazinamide. It is given by injection into a vein or muscle
MEDIUM
The culture medium for streptomycin consists of –
1. Carbon source : starch, dextrin, glucose,
2. Nitrogen source : natural agricultural by-products, soybean meal, corn steep
liquor, cotton seed flour, casein hydrolyte, or yeast & its extract. Inorganic N salts
like ammonium sulphate & ammonium nitrates are also used

13. • Inoculum -S. griseus spores maintained in soil stocks or lyophilized in
carrier are inoculated into sporulation medium, which builds up mycelial
inoculum.
• THE HOCKENHUL MEDIUM----Glucose, extracted soya meal, distillers
dried soluble sodium chloride, pH as 2.5% 4%, 0.5%, 0.25%, 7.3 - 7.5
respectively.
• Spores of S. griseus are inoculated into a medium to establish a culture
with high mycelial biomass
• • Yield in production vessel responds to high aeration & agitation
conditions.
• Other conditions involve-Temperature range 25-30°C pH range 7-8 Time
5-7 days
• Mycelium is separated from broth by filteration & streptomycin is
recovered. Recovery process - broth is acidified, filtered & neutralized

14. TETRACYCLINE
• A family of broad-spectrum antibiotics effective against a remarkably
wide variety of organisms. Bacteria susceptible to tetracycline include
Haemophilus influenzae, Streptococcus pneumoniae, Mycoplasma
pneumoniae, Chlamydia psittaci, Chlamydia trachomatis, and Neisseria
gonorrhoeae.
• The first member of the group to be discovered is chlortetracycline
(Aureomycin) in the late 1940s by Benjamin Minge Duggar, who derived
the substance from a golden-colored, fungus-like, soil-dwelling bacterium
named Streptomyces aureofaciens.

17. GRISEOFULVIN
• It is an antifungal antibiotic first isolated from a Penicillium species in
1939. It is a secondary metabolite produce by the fungus Penicillium
griseofulvum.
• Griseofulvin inhibit fungal cell mitosis and nuclear acid synthesis
• It is used in the treatment of Ringworm of the Beard, Ringworm of Scalp,
Fungal Disease of the Nails, Ringworm of Groin Area, Athlete's Foot,
Ringworm of the Body.

18. PREPARATION OF MEDIA
• Medium- Czapek Dox Medium ----Chemicals - Glucose 5%, Sodium Nitrate
0.2% , Potassium Hydrogen Phosphate 0.1%, Magnesium Sulphate 7H20
0.05%
Steps Involved In The Manufacturing Process
• Fermentation
• Pre treatment of fermentation broth
• Filtration
• Extraction
• Decolorization
• Isolation and separation
• Precipitation and purification