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The objective of the current study was to evaluate the use of a novel species of a biosurfactant-producing bacterium Pseudoclavibacter sp. strain SKC/XLW-1 as potential reagents for bioflotation of sulfide minerals (herein pyrite) in order to discover environmentally friendly flotation reagents. The bacterial strain was able to change surface chemical properties of pyrite as evidenced by Fourier transform infrared spectroscopy (FTIR) analysis and hydrophobicity measurements (contact angle and surface tension). Over the course of the experiment the contact angle tended to increase whereas the surface tension was observed to decrease as a function of time, suggesting the possible potential of the strain to function as both frother and collector for flotation of sulfide minerals. From the findings of this study it is evident that the bacterium Pseudoclavibacter sp. strain SKC/XLW-1 can be employed to be a more eco-friendly reagent (both collectors and frother) to facilitate the selective separation of sulfide minerals in the flotation processes.