Chrysosporium tropicum, renowned for its distinctive morphological traits and potential for enzymatic activity, serves as a compelling subject for exploring keratinase production and growth optimization. This study delves into the determination of optimal conditions for keratinase production by Chrysosporium tropicum and investigates the correlation between its morphological characteristics and enzymatic activity. Through a series of experiments, the effects of incubation period, temperature, and substrate amount on keratinase production were evaluated. Keratinase activity was measured using a spectrophotometer, revealing significant variations across different conditions. Results indicate that the highest keratinase activity for Chrysosporium tropicum was achieved after 16 days of incubation at 25°C, with a substrate amount of 6g. Subsequent purification of the enzyme was conducted via dialysis and column chromatography, resulting in enhanced enzyme activity. The findings underscore the biotechnological potential of Chrysosporium tropicum in keratin degradation processes and contribute to our understanding of optimizing enzyme production for various applications.