Production of streptomycin

1. Reaccredited with A+ grade with a CGPA of 3.39 in the III cycle of NAAC
Affiliated to Manomanium Sundaranar University, Tirunelveli.
Post graduate & Research Centre – Department of Microbiology
(government aided)
IV SEM CORE: FEREMENTATION AND INDUSTRIAL MICROBIOLOGY
UNIT-3
PRODUCTION OF STREPTOMYCIN
ARULSELVAM.A SUBMITTED TO
REG NO:20211232516105 GUIDE:DR.S.VISWANATHAN
II M.SC MICROBIOLOGY ASSISTANT PROFFESSOR&HEAD
ASSIGNED ON: SPKC - ALWARKURUCHI

2. Introduction :
• Streptomycin was developed by
Schatz Bugie and Waksman in 1944 .
• According to the World Health Organisation (WHO), streptomycin
is the safest medicine for treating tuberculosis. Streptomycin has
been included in the list of essential medicine needed for the health
system by the WHO. In the year 1952, Waksman won Nobel Prize
in physiology and medicine for the discovery of streptomycin.

3. Organism Required for Streptomycin Production :
• Streptococcus griseus is an actinomycete, which is used for the preparation of streptomycin.
• Krinsky was the first scientist isolated S.griseus during the time of 1st world war from the Russian soil.
• S.griseus is commonly present in the soil, and it is gram-negative bacteria. It produces grey mycelium during sporulation
and produces grey yellow reverse pigment when it grows in the form colony.

4. Streptomycin :
• Streptomycin refers to the broad spectrum antibiotic.
• Oligosaccharide antibiotic belongs to aminoglycoside antibiotics family.
• Streptomycin is bactericidal
• It is active against both gram-positive and gram-negative bacteria.
• Streptomycin is inhibitory for several species of Mycobacterium and is an effective antibiotic for treatment of
tuberculosis caused by M. tuberculosis.

5. Structure of Streptomycin :

6. It consists of three components
(i) Streptidine (inositol with two guanido groups)
(ii) Streptose (methyl pentose), and
(iii) Streptoscamine (N-methyl-L-glycosamine) .
• It has the chemical formula C21H39N7O12 .
• Both guanido groups of streptidine are essential for the antibiotic activity. Removal of one group reduces
activity up to 90%.

7. Mechanism of action :
• Streptomycin is a protein synthesis inhibitor.
• It binds to the small 16S rRNA of the 30S subunit of the bacterial ribosome, interfering with the binding of formyl-
methionyl-tRNA to the30S subunit.
• This leads to codon misreading, eventual inhibition of protein synthesis and ultimately death of microbial cells.
Preparation Of The Medium :
The production of streptomycin requires Hockenhull nutrient medium with the following media composition:
Glucose: 2.5%
Soy meal: 4%
Sodium chloride: 0.25%
Distillers dried soluble: 0.5%
pH: 7.6-8.0

8. Fermentation process :
• Sterilize all the above contents and fed into the stirred tank fermentor .
• Inoculum require - 4-5%
• Temperature - 25-30 degrees Celsius
• pH - 7.6-8
• The production of streptomycin requires continuous fermentation
• The growth of S.griseus requires continuous aeration and agitation.
• The fermentation process takes 4-10 days.
• The fermentation process involves three consecutive phases.

9. First phase :
During the first phase, the S.griseus grows by utilizing soya 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. In
this phase, the incubation period lasts for 24 hours to 6-7 days. The pH during this phase remains constant between 7.6
and 8. There is little or no production of mycelium.
Third phase :
It is the final phase of fermentation, where the concentration of glucose depletes from the medium that results in
cessation of streptomycin production, and the bacterial cells begin to lyse. As a result, the medium’s pH increases by the
deposition of ammonia after the cell lysis. Before cell lysis, harvest the fermentative medium for the process of recovery
and purification.

11. Recovery of Streptomycin :
The recovery of streptomycin includes the following stages:
Filtration:
In the recovery of streptomycin, separate the bacterial biomass from the fermentative broth through Oliver’s pre-coated
pressure filter. After filtration, the remaining liquid is first acidified, filtered and then neutralized.
Adsorption:
Mix the activated carbon with the clear broth obtained after filtration in a series of three adsorption tanks. Separate
streptomycin from the broth to remove impurities. Then, the broth is fed to the pressure filter, which releases out the
spent broth. Wash the absorbate with dilute alcohol to remove impurities.
Elution:
Elute the streptomycin via ion exchange method by adding dilute hydrochloric acid. In this process, the impurities like
metallic salts are removed from the solution, which can contaminate the streptomycin.
Concentration:
Concentrate the remaining solution at 60 degrees Celsius under vacumm by the series of three single-phase evaporators.
Crystallization: Firstly, dissolve the streptomycin in methanol and then filter it again. Then, treat the filtrate with acetone
to obtain a precipitate. Rewash the precipitate with the acetone and later dry it under vacumm.

12. Final operations :
Then, dissolve the crystalline streptomycin in the solution containing pyrogen-free water. Pass the solution through the
special biological filter like Seitz filter to eliminate the microorganisms that can contaminate the streptomycin
Uses of Streptomycin
Clinical science:
Streptomycin is very effective against gram-negative bacteria, thus treating the infections and diseases caused by gram-
negative bacteria.It is widely used in the treatment of severe disease like tuberculosis.Streptomycin is used in conjugation
with antimycobacterial drugs, to treat pulmonary infections caused by the Mycobacterium avium complex.
Patients suffering from endocarditis caused by Enterococcus can be treated with streptomycin if they are sensitive to the
gentamicin. Tularemia infections, Plague, Brucellosis etc. can also be treated with the streptomycin.
Agriculture science: Streptomycin is used as “Pesticide” that inhibits the growth of bacteria, fungi, algae etc.
Cell culturing: It is used at the time of cell culture, to prevent undesired microorganisms’ growth.
Biochemistry: Streptomycin sulphate is also used in the purification of biomolecules like protein and sometimes nucleic
acids

13. References :
 https://biologyreader.com/production-of-streptomycin.html
 https://microbiologynotes.org/streptomycin-chemical-structure-production-recovery-and-uses/
 An Introduction to Industrial Microbiology K Sukesh · 2010
 Fermentation Microbiology and Biotechnology E. M. T. El-Mansi, C. F. A. Bryce, Arnold L. Demain · 2011

16. THANK YOU
The Chairman.
The secretory.
Management committee.
The principal, Sri paramakalyani college.
The head-Department of microbiology.
The staff members- Department of microbiology.