The production of cutinase by solid-state fermentation, using by-products such as wheat bran, rice bran, or soybean rind, was carried out using a Fusarium oxysporum strain. The fermentation process was optimized using a central composite design. The best conditions for cutinase production were achieved at 28-30 A degrees C, with water added at 100-150% (w/w) after 72 h of incubation, in the range of 11.7-15.5 U/mL. In addition, the resolution of (R,S)-2-octanol and (R,S)-ibuprofen was performed to evaluate the enantioselectivity of the preparations of cutinase. The cutinase produced from the soybean rind attained higher yields and enantioselectivity in the resolution of (R,S)-2-octanol with octanoic acid in isooctane (E = 9.6). For the (R,S)-ibuprofen resolution, the cutinase produced from rice bran reached the best yields (E = 5.6). This work demonstrated that the enzymes can be produced from different media, such as from by-products or residues rich in carbon sources that do not necessarily present the same biochemical properties, which may be useful for industrial applications.