production of streptomycin bioprocess technology

1. Production of Streptomycin
BY MOHIT KOHLI

2. Streptomycin
• Broad spectrum antibiotic
• Oligosaccharide antibiotic/ aminoglycoside family
• Discovered by Schatz, Bugie, and Waksman in 1944 from Streptomyces
griseus isolated from soil
• First effective treatment against Mycobacterium tuberculosis.
• Also effective against some Gram positive bacteria primarily Staphylococcus
• Active against Pasteurella pestis, Brucella abortus, Francicella tularieusis
• It is protein synthesis inhibitor: acts by binding to the small 16S rRNA of the
30S subunit of the bacterial ribosome, interfering with the binding of
formyl-methionyl-tRNA to the 30S subunit, leading to codon
misreading followed by partial or complete inhibition of protein
synthesis

3. •Streptomycin has three constituents namely; N-methyl L-glucosamine, Streptose and
streptidine
• Present day Streptomycin producers are the mutants of Streptomyces derived for higher
yields with yield as high as 25,000 units per ml
• Industrial production of streptomycin is carried out using submerged fermentation
processes
•Streptomyces mutants are genetically unstable, the spores are maintained as soil stocks or
lyophilized and are used for inoculating sporulation medium, which is then transferred to
germinator where biomass is increased for inoculating fermenters
•Fermentation media consists of glucose, starch, dextrin, soy meal, corn steep liquor,
Sodium sulphate. The streptomycin fermentation requires high aeration and agitation.
•Fermentation is carried out at 28-30ºC with pH maintained at 7.6-8 for good productivity
The fermentation lasts for 5-7 days with of yield of 1-3 g/L of the fermentation broth

6. The streptomycin fermentation proceeds through three phases:
1. In first phase, the organism produces proteases which digest the soybean meal
and release ammonia and carbohydrates. These are utilized for increasing the
biomass. Glucose is slowly utilized and net production of streptomycin is low
during this phase. The pH of the medium increases from 6.7 or 6.8 to 7.5 or
higher. This phase lasts for 24h.
2. The next phase is the idiophase or the stationary phase during which maximum
streptomycin (secondary metabolite) is produced. It ranges from 24h to 6-7 days.
Rapid utilization of ammonia and glucose occurs with no mycelial growth and pH
during this phase remains fairly constant at 7.6 to 9.0.
3. In the last phase (death phase) the sugars have been completely depleted in the
medium and streptomycin production ceases completely. The ammonia released
due to the cell

8. Recovery of Streptomycin
• On completion of fermentation, the mycelium is separated from the broth by filtration.
• Remaining liquid is then percolated through cation exchange resin columns where
streptomycin gets adsorbed and is finally eluted by washing with buffer as
streptomycin sulphate
•Further impurities are removed by treating it with sodium hypochlorite, EDTA and
activated carbon
• Purified streptomycin sulphate solution is concentrated under vacuum and dried
aseptically

9. Kind of Vessel: Process Control etc for Streptomycin production

12. 1. Saval et al., 1993: Optimized media for streptomycin production for four components
Resulting in 52 % increase in streptomycin yield
2. Pitting to a depth of 7 mm was found in a mild steel fermentor after 7 years use for
Streptomycin production (Walker and Holdsworth, 1958)
3.Weak acid cation exchange can be used in isolation and purification of streptomycin

13. • During recovery of streptomycin, the harvested filtrate is fed on to a column of a weak
weak acid cationic resin such as Amberlite IRC 50 (Sodium form).
Flow rates between 10-30 bed volumes per hour is maintained. The resin is then rinsed
with water and eluted with dilute hydrochloric acid
After this the resin column is generated to sodium form by passing adequate volume of
NaOH.
Resin can have capacity of 1 g of streptomycin per gram resin.