AN ANTIMICROBIAL PHTHALATE DERIVATIVE FROM BACILLUS CEREUS, THE SYMBIOTIC BACTERIUM ASSOCIATED WITH A NOVEL ENTOMOPATHOGENIC NEMATODE, RHABDITIS (OSCHEIUS) SP
Objective: To isolate and identify the bioactive metabolites from the culture filtrates of a bacterium (Bacillus cereus) symbiotically associated with a novel entomopathogenic nematode Rhabditis (Oscheius) species.
Methods: The bacterium was cultured in three different media and the antimicrobial activity was determined by the well diffusion assay. The ethyl acetate extract of the cell free culture filtrate was then purified by silica gel column chromatography and thin layer chromatography. Identification of the active metabolite was done with HPLC, GC-MS and LC-MS.
Results: The cell free culture filtrate of a nematode symbiotic bacterium showed both antibacterial and antifungal activities. Fermentation conditions were standardized and optimum antibacterial activity was observed in tryptic soy broth at 72 h of incubation at 30 Â°C. When the ethyl acetate extract was purified by silica gel column chromatography and thin layer chromatography, an active fraction was obtained which was subjected to HPLC analysis along with GC-MS and LC-MS leading to the identification of a major compound Bis (2-ethyl hexyl) phthalate. The compound was active against Gram positive bacteria Bacillus subtilis MTCC2756, Staphylococus aureus MTCC902, Gram negative bacteria Escherichia coli MTCC 2622 and fungi such as Aspergillus flavus MTCC277, Candida albicans MTCC183, Fusarium oxysporum MTCC 284, Rhizoctonia solani MTCC 4634.
Conclusion: Bis (2-ethyl hexyl) phthalate was identified as one of the metabolites produced by a nematode symbiotic bacterium associated with a novel entomopathogenic nematode Rhabditis (Oscheius) species. Thus similar compounds isolated from novel entomopathogenic bacteria would pave the way for identifying new drugs for the pharmaceutical and agricultural sector.
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