Starch nanoparticles (SNPs) in colloidal forms were prepared using enzymatically pretreatment and four different gelatinization methods based on autoclave, microwave, ultrasonication and normal heating with stirring. Furthermore, SNPs in powder form were prepared using freeze drying technique. Results indicated that the formed SNPs using starch solution (1 % W/V) and ultrasonication technique had lowest mean particle size (151 nm) and PDI (0.173), and highest zeta potential (−8.8 mV) values. Optimization procedure using response surface methodology, based on central composite design, indicated that using 1.5 mL of α-amylase and sonication time of 15 min, SNPs with lowest particle size (49.3 nm) and highest zeta potential (−10.8 mV) were produced. Using prepared colloidal solution under optimal conditions, SNPs powder were produced by freeze dryer, adjusted at pressure and temperature of 100 Pa and − 70 °C, for 24 h. Results indicated that formed SNPs powder with squared-shape, had particle size, zeta potential, specific surface area, decomposition temperature of 197 nm, −13.9 mV, 1.9 m2g−1 and 162 °C, respectively. While, for native starch these values were 5018 nm, −6.01 mV, 0.68 m2g−1 and 170.2 °C, respectively. Results revealed that emulsification ability of SNPs powder was three times higher than that of the native starch.