It is about information about well known pharmaceutical industry in Pune , which also includes production of penicillin and Streptomycin which is helpful for biotechnology students

1. Drug manufacturing company
HINDUSTAN ANTIBIOTICS LIMITED

2. INTRODUCTION
• Known as ‘penicillin factory’ situated at
Pimpari, Pune and set up on 10th March,
1954.
• The public sector company by Government
of India with active co-operation of WHO
and UNICEF.
• Board of Director - Ms. Nirja Saraf.
• To undertake commercial production of
antibiotics like Penicillin, Streptomycin,
Gentamicin, Ampicillin & Amoxycillin etc.
• Company has facility to manufacture
various dosage forms - injectables,
capsules, tablets, large volume parentrals,
liquid orals, etc.

3. MISSION
• To Manufacture products of highest
quality standards .
• To Follow current Good Manufacturing
Practices (cGMP), Good Laboratory
Practices (GLP) .
• To Ensure the products manufactured
and distributed fulfil to National and
International Standards of quality,
purity, efficacy & safety.”
• Company’s vision –
“Cure for Millions & Care for All.”

4. YEAR MILESTONES
1954 Production of Potassium Penicillin 1st Crystals.
1956 Factory inaugurated by Pt. Jawaharlal Nehru, Prime Minister of India.
1962 Streptomycin plant Started at commercial basis.
1975 Production of Ampicillin started for the first time in India on the commercial basis.
1978 Gentamicin First batch of Gentamicin Injectable started.
1987 Production value increased by 40 percent.
1988 Increased Penicillin capacity to 360 MMU.
1992 Increased of Penicillin capacity to 900 MMU.
2009 Production of Cephalosporin started .
2018 Achieved the highest value of production and sales for Streptomycin.
MILESTONES

5. PRODUCTS
PENICILLIN -
• Antibiotic obtained
from penicillium
mould
• Treat bacterial
infections , mostly
pneumonia.
STREPTOMYCIN -
• Aminoglycoside
• Obtained from
Streptomyces
• Treat tuberculosis,
Mycobacterium avium
complex, endocarditis,
brucellosis.
GENTAMICIN –
• Aminoglycoside
antibiotic
• Works by killing
bacteria or preventing
their growth.
• Treat bacterial
infections.
AMPICILLIN –
• Penicillin group of
beta- lactam
antibiotic
• Treat meningitis and
infections of the
throat, sinuses, lungs.
AMOXYCILLIN –
• Newer version of
penicillin that treat
more types of
bacterial infection.
• More potent –
contain modify
structure of penicillin.

6. • Penicillin is first antibiotic to be
discovered.
• In 1928, Alexander Fleming discovered
that mould from penicillium fungus
had antimicrobial properties.
• Penicillium is broad range antibiotic
which is used to treat wide range of
infections and diseases, including
streptococcus, scarlet fever, syphilis
and gonorrhoea.
• Penicillium is secondary metabolite
produced by microorganisms during
stationary phase.
PENICILLIN
Penicillium mould on bacterial
petri plate

7. INDUSTRIAL PRODUCTION – PENICILLIN
• From Penicillium chrysogenum.
• Fed batch culture- A fed batch culture is fed
continuously without removal of culture fluid.
• Penicillin notatum and Penicillin chrysogenum are
two species widely used in fermentation.
• Penicillin chrysogenum is widely used as it is high
yielding strain.
• It include upstream and downstream processes.
• Upstream process- Development and production.
• Downstream process- Extraction and purification.

8. Layout
Layout of production of penicillin by Fed batch method

9. INOCULUM PREPARATION-
• Lyophilized spores transferred
into agar plate(to allow growth).
• Spores start geminate and form
mycelia.
• Then culture incubated for 70
hours in rotary shaker at 200
rpm and 25 degree Celsius.
• Resulting spores transferred into
seed tank

10. 2)TRANSFER OF INOCULUM INTO SEED TANK
Seed fermenter -
1)Smaller in size than main fermenter
2)Two liter capacity
3)Made of stainless steel
4)Equipped with agitators(allow mixing of
medium)
5)Pump is there to delivered sterilized air.
Initial stages of fermentation designed for
considerable microbial growth. To support
mycelium growth shift prepared culture to
seed tank/seed fermenter.
Keep prepared culture under aeration and
agitation for 24-48 hours.
This stage is known as “Growth phase of
mould”
Seed fermenter

11. 3) FERMENTATION
• Fed batch culture- A fed batch culture is fed continuously without removal of
culture fluid.
• Add inoculum into fermenter filled with production medium.
• Production medium composition:
Media composition Percentage
Lactose 3-4
Glucose (carbon source) 10
Corn steep liquor (nitrogen
source)
4
Caco3 (acts as buffer) 1
Phenyl acetic acid 0.5
Antifoam (edible oil) 0.25-0.5
Potassium dihydrogen
phosphate
0.4

12. CONTINUED
Parameters Status
PH 6.5-7
Aeration Continuous flow of
sterilized air
Agitation Baffles is present which
prevent vortex and
enhance agitation
Temperature 26-28 degree Celsius
Fermentation process requires 7 days for penicillin production. By the 7 th day PH of
medium is 8 and penicillin production stops.

13. CONTINUED
First Phase - Second Phase - Third Phase -
Growth of mycelium occurs in
this phase
Intense synthesis of
penicillin occurs due
to rapid
Concentration of antibiotics
decreases in medium
Yield of antibiotic is low Mycelium mass
increases
Autolysis of ammonia starts
PH increase due to release of
ammonia.
PH remains same. Liberation of ammonia and
slight increase in PH

14. 5) RECOVERY AND PURIFICATION:
• After penicillin fermentation, broth is filtered on rotary vacuum filter.
• Mycelia is separated from broth.
• Addition of phosphoric acid or sulphuric acid to convert penicillin into anionic
form(PH 2-2.5).
• Continuous, counter current, multistage extractors used.(to avoid degradation
temperature 2-4 degree Celsius, low PH, filtration time is very short).
• Active charcoal is added for removal of pigment and other impurities.
• Product is back extracted into water from solvent by adding sodium/potassium
hydroxide to form its salt.
• The resultant sodium/potassium penicillin is then crystalized.
• Then it is washed, dried and used for commercial purpose.

15. INDUSTRIAL PRODUCTION - STREPTOMYCIN
• Streptomycin is an antibiotic that act against the gram-negative bacteria as well as gram
positive bacteria.
• It is the first antibiotic to treat tuberculosis.
• Streptomycin first isolated by scientist Albert Schatz from Rutgers University in 1943.
• Streptomycin is produced by submerged culture method at industrial or commercial level.
1)
• INOCULUM PRODUCTION
2)
• PREPARATION OF MEDIUM
3)
• FERMENTATION

16. LAYOUT
Layout of Streptomycin production by submerged method

17. INOCULUM PRODUCTION
Streptomyces griseus spores in the stock soil culture. Then,
inoculate the inoculum of S.griseus into the agar media at 28
degrees Celsius.
Then the spores of S.griseus sporulate and form the mycelial
structure in the flask or seed tanks.
The sporulation of S.griseus occurs well in a liquid culture
medium that is in broth , and it forms endospores in low
nutrient availability. pH range – 5-11.

18. CONTINUED
Growth of Streptomyces griseus

19. PREPARATION OF MEDIUM
• The production of streptomycin requires
Hockenhull nutrient medium.
• Carbon source: Monosaccharide like glucose
and Sugars like fructose, maltose, starch etc.
used as the carbon source.
• Nitrogen source: Components like soy meal,
meat extract, ammonium salts etc. serve as the
nitrogen source.
• Mineral source: Magnesium, calcium,
potassium etc. Provides a mineral source .
• Growth stimulating factors: L-naphthalene
acetic acid and phenyl acetic acid .
• Accelerators : Compounds like proline, oils,
fatty acids, antioxidants.
• The precursors are not required for the
production of streptomycin.
Hockenhull nutrient medium composition
Nutrient %
Glucose 2.5
Soy meal 4
Sodium Chloride 0.25
Distillers dry soluble 0.5
pH 7.6-8.0

20. FERMENTATION
Stirred tank fermenter
Sterilize all the nutrient contents and fed into the
stirred tank fermenter. Then, transfer the bacterial
inoculum of about 4-5% to the medium.
Maintain a temperature between of 25-30 degrees
Celsius and pH between 7.6-8 inside the fermenter.
The production of streptomycin requires continuous
fermentation, and the growth of S.griseus requires
continuous aeration and agitation.
The fermentation process is aerobic and takes 4-10
days. The fermentation process involves three
consecutive phases.

21. CONTINUED
FIRST PHASE - The S.griseus grows by utilizing soy meal
and releases ammonia through proteolytic activity. The pH
rises during this phase. S.griseus slowly utilizes the carbon
source present in the soy meal and the glucose and
thereby producing streptomycin at a slower rate.
SECOND PHASE - After the utilization of soy meal and
glucose, the ammonia is secreted and the streptomycin is
synthesized rapidly. Incubation - 24 hours to 6-7 days. The
pH remains constant between 7.6 and 8. There is little or
no production of mycelium.
THIRD PHASE - The concentration of glucose depletes from
the medium that results in termination of streptomycin
production and the bacterial cells begin to lyse. Before cell
lysis, harvest the fermentative medium for the process of
recovery and purification.

22. RECOVERY AND PURIFICATION
FILTRATION ADSORPTION ELUTION
CONCENTRATION
EVAPORATION
Crystallization
Final
operations
FERMENTATION
BROTH
Final product – STREPTOMYCIN
By-product – VITAMIN B12

23. ACHIEVEMENTS AND AWARDS
• HAL has achieved an all
time high sales of 59.92
Lacs pouches (6 grams)
of Streptocycline during
the year 2017-18.
• HAL’s R & D has the
distinction of being the
only laboratory in the
public sector in India
which has come out with
its own discoveries of new
molecules Viz. Hamycin
for skin infections and
Aureofungin for plant
fungal control.
• Special Products: HAL has
to its credit production of
exclusive products like
“Sodium Chloride with
Glycerine Injection” and
“Potassium Iodate
Tablets” for Defence
Sector and “Narcotics
Detection Kits” for security
applications.
• HAL has significantly
improved its performance
during the year 2017-18
with the value of
production.

24. SOURCES
 https://biologyreader.com/wp-
content/uploads/2020/04/production-process-of-penicillin.jpg
 https://th.bing.com/th/id/OIP.fw8Aqj-
BAeEs_uIrw8UmKgAAAA?pid=ImgDet&rs=1
 https://www.researchgate.net/profile/Marijke-
Luttik/publication/7512467/figure/fig1/AS:814960196190212@1
571313060890/Conidial-growth-of-Penicillium-chrysogenum-Image-
kindly-made-available-by-CBS-Fungal_Q320.jpg
 Industrial Production of Penicillin (learninsta.com)
 https://www.news-medical.net/health/Penicillin-Production.
 https://www.slideshare.ne
 Production of Penicillin? Overview, Industrial Production & Uses -
Biology Reader