Abstract: Background and Objective: Fungal pathogens develop resistance against antifungal agents available in market. Therefore, treatment of fungal infection is becoming problematic. On other hand, chemically synthesized antifungal agents are also causing adverse effect on human health. To overcome this issue, recently many drug industries are trying to isolate bioactive molecules from natural sources. The objective of the study was isolation and identification of bacterial isolate from contaminated agar plate with special reference to its antifungal property and explore bacterial contamination as new habitat for beneficial microbes. Materials and Methods: Antifungal metabolite producer Pseudomonas tolaasii (P. tolaasii) GD76 was isolated from contaminated yeast, peptone, dextrose (YPD) agar plate on to which it was streaked from casein agar plate. Morphological, biochemical and phylogenetic study was carried out for identification of the bacterial isolate by using 16S rRNA gene sequencing and evolutionary history was inferred using the maximum parsimony method. Results: Morphological and biochemical study of the bacterial isolate showed that the isolate was Gram negative, rod shape, capsule containing and ferment only glucose. On basis of position sequence of the isolate in phylogenetic tree, the isolate showed 99% similarity to Pseudomonas tolaasii, it is indicated that the isolate is the novel strain of P. tolaasii for which the name Pseudomonas tolaasii GD76 is affirm. Sequence of 16S rRNA gene was submitted to NCBI, Accession code; KU533778. Conclusion: On the basis of results obtained, present study indicates towards the potential of uncharacterized prokaryotic community associated with bacterial contamination.