The present work is carried out to isolate antibiotic producing soil bacteria from Dibrugarh district and elucidate the structure of the metabolite. 100 samples were collected from 10 different zones of the district and serial dilution upto 10-9was done. Samples were cultured in different culture media. Total 160 isolates were isolated of which 147 are bacteria and 13 are fungi. Among all these two potentially viable isolates namely NAA and EMBA were selected for further studies. The molecular identification by sequencing of the 16S rDNA gene confirms that the isolates NAA and EMBA were strains of Enterobacter cloacae with 95% and Klebsiella pneumoniae with 92% similarity . The 16S rDNA sequence of the isolate NAA and EMBA were compared with that of other bacterial sequence by the way of BLAST and phylogeny was constructed. Our findings are in agreement with the earlier identification of Enterobacter and Klebsiella strains( Zhou et al.,2018). On the basis of sequence alignment it was found that Enterobacter cloacae culture clone CLM107 and Enterobacter cloacae strain VITADJ are 99% similar with our sequence. The selectively amplified 16S rDNA gene sequences from total genomic DNA by PCR with the help of universal primer helps in construction of phylogeny. The obtained 16S rDNA sequence were deposited in NCBI GenBank and accession numbers for NAA was KX859176 and for EMBA was KY111020. The extract of isolate NAA was found to be more active than the isolate EMBA and further LC-MS,FTIR and NMR study of the extract NAA was done. The molecular weight of the extract is 582.6. The molecule contains a furan ring which is responsible for its antimicrobial activity. In the subtropical regions of the world, around 20,000 deaths per year were due to parasitic bacterial infections(Rani et al.,2015).In spite of different available antibiotics in the market , there is an ample requirement for novel antibacterial agents in view of rapid developing of antibiotic resistant bacteria(Sekhar,2010).Keeping this in mind our finding of new molecule can serve as a new potent source of antibiotic in the field of antibiotic study.