This study investigates the effectiveness of the Process-Oriented Guided Inquiry Learning (POGIL) method compared to interactive lecturing in undergraduate biochemistry education from two private medical colleges of Punjab. The study based on quantitative assessments, including pre-tests, knowledge acquisition evaluations, and knowledge retention assessments, were conducted. Students were divided into two groups: the control group experienced interactive lecturing, while the intervention group was taught using the POGIL method. Results revealed significantly higher mean scores for knowledge acquisition in the POGIL group (M=9.18, SD=1.23) compared to the interactive lecturing group (M=7.82, SD=0.99) (p=0.05). Additionally, the POGIL group demonstrated superior long-term knowledge retention, with mean scores of M=8.89 (SD=1.23), compared to the interactive group's M=7.67 (SD=0.99) (p=0.05). These findings underscore the enhanced effectiveness of the POGIL method in fostering both knowledge acquisition and retention in enzyme biochemistry among undergraduate medical students, suggesting its potential for integration into biochemistry medical curricula.