Naringin is a major flavanone glycoside responsible for bitterness in citrus juices, particularly grapefruit juice, which often limits consumer acceptance. Enzyme-mediated hydrolysis represents a promising approach to mitigate this bitterness. Naringinase, an enzyme complex capable of catalyzing the hydrolysis of naringin, was the focus of the present study, which investigated the production potential of Penicillium roqueforti. Several culture media compositions were investigated to enhance naringinase production, and extracellular enzyme activities and pH dynamics were monitored. Based on prior studies, the initial pH values of the media were set at 4.5 and 6.5, identified as optimal for naringinase activity. The highest naringinase activity (1.47 EU/mL) was obtained in Medium 1 under different initial pH conditions. whereas variations in pH did not significantly alter the peak enzyme activity achieved in this medium. Moreover, P. roqueforti altered the pH of all media, with pH values in A1 and A2 rising to 7.86 and 8.12, respectively, by day five. A preliminary partial purification of the enzyme was performed, revealing precipitation during ammonium sulfate elution for reasons yet to be elucidated.