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
