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

Abstract

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.