Abstract
A Gram positive, filamentous, spore forming antagonistic Streptomyces sp. AP-123 derived from marine region of Andra Pradesh, India, was studied for its medical importance. Among the 210 Streptomyces strains screened at 64.3% exhibited activity against Gram positive bacteria, 48.5% showed activity towards Gram negative bacteria, 38.8% exhibited both Gram positive and negative bacteria and 80.85% strains revealed significant antifungal activity. However, primary screening revealed that Streptomyces sp. AP-123 exhibited significant antimicrobial activity against all the tested bacteria compared to other Streptomyces strains. The presence of L-diaminopimelic acid and glycine in the cell wall hydrolysates and streptomycin resistance indicated the strain belonged to Streptomyces genus. The 16S rDNA gene based phylogenetic affiliation was determined by using bioinformatic tools and it was identified as Streptomyces sp. AP-123 with 99% sequence similarity to Streptomyces flavogriseus. The antimicrobial substances were extracted by hexane and ethyl acetate from spent medium in which Streptomyces sp. AP-123 was cultivated at 30 degrees C for 5 d. The antimicrobial activity was assessed using broth micro-dilution technique. A compound was obtained by eluting the crude extract using varying concentrations of solvents followed by the chromatographic purification. Based on the IR, C-13 NMR and H-1 NMR spectral data, the compound was identified as polyketide related antibiotic. It exhibited significant antibacterial activity against Gram positive and Gram negative bacteria and also showed a potent cytotoxic activity against cell lines viz. Vero (Green monkey kidney) and HEP2 (laryngeal carcinoma cells) in vitro. The lowest Minimum Inhibitory Concentration (MIC) of the compound against Bacillus sub fills and Staphylococcus aureus were 25 and 37.5 mu g mL(-1), respectively. Against Escherichia coli and Pseudomonas aeruginosa it exhibited MIC of 50 and 37.58 mu g mL(-1), respectively. However, against Candida albicans and filamentous fungus, Aspergillus niger the MIC values were 12.5 and 25 mu g mL(-1), respectively. Cloning and sequence analysis of ketoacyl synthase (KS) gene revealed similarity to the type II polyketide synthase (PKS) gene of Streptomyces species. (C) 2012 Elsevier Ltd. All rights reserved.