Meat tenderness is one of the most important organoleptic properties in determining consumer acceptance in meat product marketability. Therefore, an effective meat tenderization method is sought after by exploring plant-derived proteolytic enzymes as meat tenderizer. In this study, a novel protease from Cashew was identified as a new alternative halal meat tenderizer. The extraction of cashew protease was optimized using response surface methodology (R2 = 0.9803) by varying pH, CaCl2 concentration, mixing time, and mass. pH 6.34, 7.92 mM CaCl2 concentration, 5.51 min mixing time, and 19.24 g sample mass were the optimal extraction conditions. There was no significant difference (n = 3; p Anacardium occidentale) crude extract can be used as an alternative of the animal and microbial protease as meat tenderizer and subsequently overcome the shortcoming of the halal industrial protease.