In this study, the methyl ester production process from neutralized waste cooking oils is optimized by using alkali-catalyzed (KOH) single-phase reaction. The optimization process is performed depending on the parameters, such as catalyst concentration, methanol/oil ratio, reaction temperature and reaction time. The optimum methyl ester conversion efficiency was 90.1% at the optimum conditions of 0.7 wt% of potassium hydroxide, 25 wt% methanol/oil ratio, 90 min reaction time and 60 degrees C reaction temperature. After the fuel characteristics of the methyl ester obtained under optimum conditions were determined, the effect on engine performance, CO and NO,, emissions of methyl ester was investigated in a diesel engine with a single cylinder and direct injection. When compared to diesel fuel, engine power and torque decreased when using methyl ester, and specific fuel consumption increased. NOx emission increases at a rate of 18.4% on average through use of methyl ester.