Corynespora cassiicola (Berk. & Curt.) Wei is an important phytopathogenic fungus, and different isolates show great diversity in their reproductive structures. Therefore, the aim of this study was to evaluate the potential of the API-ZYM® system as an auxiliary tool in the polyphasic approach of C. cassiicola identification. Five C. cassiicola isolates from different host plants and one Pseudocercospora griseola isolate were tested. A typical enzymatic pattern was obtained, with eight enzymes being produced by all five C. cassiicola isolates. An intraspecific differentiation was also found. Certain enzymes produced, such as α-glucosidase, β-glucosidase and α-mannosidase, may be related to pathogenic processes, acting in the degradation of the structural components of the host cells.