Sugarcane (Saccharum officinarum L) is squeezed to obtain juice after being purified and neutralized, allowed to crystallize the sugar. Considering the importance of sugar cane accounts for the sugar industry, it is necessary to increase the content of dextran-free sucrose (sucrose destruction caused by the action of microorganisms accompanying the cane, one of the most important Leuconostoc mesenteroides). In this research we determined the inhibition of lactic acid bacteria L mesenteroides isolated from a sugar mill in Valle del Cauca using antagonist microorganisms. For this, isolates, and cultures of L. mesenteroides from cane juice and performed various bio-chemical and microbiological tests in the laboratory for isolation and identification of both strains of Leuconostoc mesenteroides as the bacterial strains antagonistic to it. It was found that strains of the fungus Trichoderma sp, Metharhizium anisopliae, were the most efficient in controlling growth of L. mesenteroides, showing a significantly higher percentage of inhibition compared to the filamentous fungi Colletotrichum sp, Thichoderma viridae, Rhizoctonia sp, and Bacillus subtilis and Serratia bacteria and yeasts marcenses Rhodothorula Rubra and Kloeckera japonica, showed no growth inhibition L. mesenteroides. In addition, we identified the fungus Aspergillus sp, Botrytis sp and Streptomyces sp as natural enemies and common in the juice of the Sugar Cane.