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PRODUCTION, PURIFICATION

AND PRODUCT RECOVERY OF


STREPTOMYCIN
PRESENTED BY
SUMAN RAI
CCT, DHARAN
INTRODUCTION
Antibiotics are secondary metabolites produced
at the end of log phase.
They are the agents that are produced by one
microorganisms and kills or inhibit the growth
of other organisms.
The term antibiotic was first used in 1942 by
Selman Waksman.
Penicillin was the first antibiotic discovered by
Alexander Fleming in 1929 from PenicilliIum
notatum.
Most commonly used antibiotics are :
Penicillin, Streptomycin, Ampicillin, Kanamycin,
Chloramphenicol, etc..
Antibiotics are produced industrially by a
process of fermentation.
STREPTOMYCIN
Aminoglycoside antibiotic.
It was discovered by Selman Waksman in
1944.
It is active against gram negative bacteria and
tuberculosis organism Mycobacterium tuberculosis.
Very less effective against gram positive but
still used in gram positive penicillin resistance
bacteria.
Streptomycin is a protein synthesis inhibitor.
It is produced by several species of
microorganisms but commonly, Streptomyces
griseus, S bikiniensis, S homidus etc are used
commonly.
• Molecular structure:
C21H39N7O12
PRODUCTION OF STREPTOMYCIN
• Medium for growth: Medium for the
fermentative process of Streptomycin production
essentially compromises of:
I. Carbon sources: E.g. dextrin, glycerol, glucose,
starch.
II. Nitrogen source: E.g. cotton seed meal,
soybean meal, casein hydrolysate etc.
III. vegetable/ Animal fat: E.g. soybean oil, linseed
oil.
Parameters:
Temperature : between 25-30° C (~ 28 °C)
pH : Ranges between 7.6-8
Duration : 5-7 days
( Yield >1200 mcg/ml )
Streptomycin hardly gets destroyed by the
presence of contaminating microorganisms.
PROCESS
Production phases
Phase 1
Phase 2
Phase 3
• Purification phase and product recovery
Phase 1
 Extends up to only 24 hours
 Produces a large proportion of mycelium
 Glucose up-take of the medium is very low
(which may be the reason for low production).
 Strong proteolytic activity of organisms
releases ammonia from soybean meal.
 The pH rises to about 8.0 due to the
production of ammonia.
Phase 2
Most crucial and critical stage
Streptomycin is generated at high rate
Extends from 1 day to almost 6/7 days of
incubation
The ammonia is utilized and pH falls to 7.0
Glucose also being used-up to the maximum
extent.
Phase 3
 The mycelium undergoes autolysis with the
rise in pH value.
 After a period of continued streptomycin
production, the pH falls again.
 Reduction in the oxygen requirement and
the content of the medium is exhausted.
Purification and product recovery
Fermented liquid is filtered to remove the
mycelium.
Thus streptomycin is finally recovered by one
of the two methods as shown below:
Method 1
Mycelia is separated from fermented broth
by filtration in rotatory vaccum filter.
Clear broth is then treated with activated
carbon particles.
Elution from carbon with dilute acid
Solvent extraction and precipitation
Filtration to get precipitates
Dried
Further purification before distribution and
use.
Method 2
i. Clear broth
ii. Acidified , filtered and neutralised
iii. Passed through column containing cation
exchange resin
iv. Resin adsorb antibiotics
v. Column washed with water to remove
impurities
vi. Elution with 0.1 N HCl
vii. Removal of water by drying
viii. Dissolve in water and methanol
ix. Addition of acetone for precipitation
x. Vacuum drying
xi. Dissolves in methanol
xii. Add CaCl2
xiii. Streptomycin chloride salt
• The final product must meet standards set by
the Food and Drug Administration.
REFRENCES
• Hockenhull, D. J. D. 1960. The biochemistry of streptomycin
production. Pp. 131-165 in Progress in industrial microbiology,
vol. II (ed, by D. J. D. Hockenhull). Intersience Publishers, Inc.,
New York.
• ^ Raymon, Lionel P (2011) COMLEX Level 1 Pharmacology
Lecture Notes, Miami, FL Kaplan, Inc. p. 181. CM4024K
• ^ http:// antibiotics toku-e. com/antimicrobial_1099_1 html
THANK YOU