The present study optimized the submerged fermentation conditions of Pediococcus pentosaceus Sanna 14 culture to improve bacteriocin yield by applying response surface methodology (RSM) and hybrid artificial neural networkgenetic algorithm (ANN-GA). A full factorial central composite design (CCD) of RSM was applied to assess the effect of four principle variables, i.e., pH (4.0-8.0), agitation (120-220 rpm), sucrose (20-40 g/l), and peptone (5-20 g/l), on the yield of bacteriocin. The RSM optimized the experimental results of pH (7.0), agitation (200), sucrose (40 g/l), and peptone (20 g/l), and supported a higher yield (2.4 g/l) of bacteriocin and was validated applying ANN-GA methodology. The RSM bacteriocin yield (2.4 mg/l) was found to match with the ANN-predicted yield (2.4 mg/l). GA results confirmed the genetic fitness of the culture of P. pentosaceus Sanna 14 during fermentation. The present study registered a sixfold increase in bacteriocin yield (2.4 mg/l) compared to the yield (0.4 mg/l) of the unoptimized process conditions.