Djulis (Chenopodium formosanum Koidz.) is an annual fast-growing underutilized pseudo cereal with a high percentage of starch content. In this study, djulis starch was extracted from the flour of dried grains by three different isolation procedures: (1) hydrochloric acid (HCl) isolation procedure (HP); (2) deionized water isolation procedure (WP); and (3) sodium hydroxide (NaOH) isolation procedure (NP), followed by investigation of the physicochemical properties of the isolated djulis starch. The amylose content of HP, WP, and NP was 22.14%, 24.15%, and 22.43%, respectively. For scanning electron microscopy (SEM) morphological observation, djulis starch presented a polygonal shape with granule sizes of 0.56–1.96, 0.74–3.02, and 0.62–2.48 μm, respectively. Djulis starch showed the classification of typical A-type x-ray patterns, and the relative degree of crystallinity for HP, WP, and NP was 33.15%, 36.17%, and 37.42%, respectively. Differential scanning calorimetry (DSC) analysis was used to determine the transition temperatures, transition range, and enthalpies of the gelatinization of starches. HP and WP isolated starch exhibited the highest ΔH 9.24 and 8.51 J/g, respectively, whereas NP starch showed the lowest ΔH of 6.95 J/g. The pasting temperatures of HP, WP, and NP isolated starch, which were analyzed by using a Rapid Visco Analyzer (RVA), were 71.70 °C, 72.80 °C, and 69.53 °C, respectively. The dependence of swelling power for the three isolated starches on temperature was tested at 10 °C with intervals between 60 °C and 90 °C. In short, the NP isolation procedure with a stable reaction is compelling from a technological point of view.