Biochemical studies on the production of Sparsomycin antibiotic by Pseudomonas aeurginosa,AZ-SH-B8 using plastic wastes as fermented substrate

Hundred and twenty microbial isolates could be isolated from different soil samples collected from different localities in Egypt. One of the bacterial cultures AZ-SH-B8 was found to produce a wide spectrum antimicrobial agent when cultivated on plastic wastes. The bacterial culture AZ-SH-B8 could be isolated from a soil sample collected from Sharkia governorate, Egypt. From the taxonomic features, the bacterial isolate AZ-SH-B8 matches with Pseudomonas aeurginosa in the morphological, physiological and biochemical characters and confirmation by using API20E system. Thus, it was given the suggested name P. aeurginosa, AZ-SH-B8. The parameters controlling the biosynthetic process of antimicrobial agent formation includes: innoculum size, different pH values, different temperatures, different incubation period, different carbon and nitrogen sources and different mineral salts concentrations (KNO₃, K₂HPO₄, MgSO₄·7H₂O and KCl) were fully investigated. The active metabolite was extracted using n-butanol (1:1, v/v) at pH 7.0. The separation of the active ingredient and its purification was performed using both thin layer chromatography (TLC) and column chromatography (CC) techniques. The physico-chemical characteristics of the purified antibiotic viz. color, melting point, solubility, elemental analysis, spectroscopic characteristics and chemical reactions have been investigated. This analysis indicates a suggested empirical formula of C₁₃H₂₁N₃O₆S₂. The minimum inhibition concentrations “MICs” of the purified antimicrobial agent were also determined. The purified antimicrobial agent was suggestive of belonging to Sparsomycin antibiotic produced by P. aeurginosa, AZ-SH-B8.