Purification and Identification of Extracellular Secondary Metabolites from Actinomycetes and Study Its Antibacterial Activity Against Pathogenic Bacteria

Document Type : New and original researches in the field of Microbiology.

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Department of Biology, College of Science, University of Babylon, Babylon, Iraq

Abstract

Background: Actinomycetes are Gram-positive, spore -forming filamentous bacteria known for producing a broad spectrum of antibiotics compounds. Objective: This study aimed to isolate Actinomycetes from soil samples collected across Babylon province and assess their antibacterial potential against multidrug resistant bacteria. Methodology: A total 100 soil samples were treated with calcium carbonate (CaCO3), serially diluted, and cultured on ISP2 medium selective for Actinomycetes. The spore morphology was examined use electron microscope. Secondary metabolites were extracted using ethyl acetate, partially purified by thin-layer chromatography (TLC), and analyzed by high -performance liquid chromatography (HPLC) with a photodiode array detector. Results: The chromatographic profile revealed multiple peaks in the crude extract, with highest peaks intensity observed between 2.00 to 5.30 minutes. Antibacterial activity was evaluated using the agar well diffusion method against multidrug -resistant bacterial isolates, including 13 strains of Staphylococcus aureus, 13 strains of Klebsela pneumoniae and 14 strains of Escherichia coli.The extract showed inhibition zone from 4.00 to43.00 mm across concentrations of 25,50,100 and 200 mg/ml. Statistically significant results (p<0.05) were observed,  with the highest efficacy against S.aureus. Conclusion: This findings confirm that secondary metabolites were derived from Actinomycetes possess significant antibacterial properties especially against multidrug-resistant S.aureus, while showing less activity against K.pneumoniae.

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