The objective of this study was to assess the effects of lintnerization, autoclaving, lintnerization followed by autoclaving, and freeze thawing treatments on the production of resistant starch from Pathumthani 80 (RD 31) rice. The produced resistant starch was further evaluated for some important physicochemical properties including pasting properties, swelling behavior, digestibility, water holding capacity, and functional properties including glycemic index and antioxidant properties. The lintnerization treatment and autoclaving significantly (p ˂ 0.05) increased resistant starch content to 64% (w/w) and gave the lowest glycemic index (46.12%). The lintnerization followed by autoclaving treatment significantly increased the solubility and water holding capacity, reduced the swelling power, and disrupted the crystalline structure of the starch granules. The native rice starch with autoclave treatment exhibited the highest swelling power among the samples, while the acid hydrolyzed starch was followed by autoclave treatment showing the lowest swelling power (1 g/g) at 90 °C. Fourier transform infrared analysis revealed the modified structures and bonding of the starch materials with the shifting of C=O stretch. However, the antioxidant properties and pasting properties were observed to decrease with the lintnerization, autoclaving, and freeze-thawing treatment of the native starch. The highly resistant starch content and low glycemic index value of the autoclaved RD 31 starch indicates the potential of the resistant starch’s application for the formulation of functional foods targeting the diabetic population.