The interest in development of a broad-spectrum biocontrol agent against different pathogenic microorganisms is very high mainly because of public opinion. Biological control offers a friendly alternative to the use of chemical products against micotoxigenic fungi. Another reason of this concern is the selection of strains more and more resistant to the pharmaco available in the pharmaceutical industry or the inexistence of an efficient pharmaco to combat certain fungi as, for example, Cryptococcus neoformans.  Therefore, in this study, a new antimicrobial substance (AMS) produced by Paenibacillus polymyxa SCE2, capable to inhibit not only micotoxigenic fungi but also bacteria and other human pathogenic fungi were isolated. The activity of the AMS was tested against the microorganisms Micrococcus sp., Escherichia coli PCEM and Aspergillus versicolor ATCC 16853 during the isolation process. The supernatant of strain SCE2 containing the AMS was applied in two different columns (Amberlite XAD-2 and Sephadex LH-20) and the partial purified AMS was submitted to a bioauthography using the same microorganisms described above as indicators. Zones of inhibition were observed against all of them, however the inhibition zone against E. coli was observed at the origin. The same retention factor in a thin layer chromatography was observed when both Micrococcus and A. versicolor were used. When the AMS was analyzed by HPLC, four active fractions were isolated.  However, further studies as nuclear magnetic resonance and mass spectrometry are still necessary to fully characterize these substances. In addition, an ultra-structural analysis and “in vitro” antagonism study of the activity of the AMS against A. versicolor conidia and mycelium were done in liquid media. After 7h-incubation, no available cells of A. versicolor could be observed, showing the strong effect of the AMS in the growth and viability of A. versicolor.