| Summary: | <i>Hodgsonia heteroclita</i> subsp. <i>Indochinensis</i> W.J.de Wlide & Duyfjes (or Making in Thai) is a neglected and underutilized crop (NUC) with high fat containing nuts. In this study, the enzyme-assisted mechanical extraction of <i>H. heteroclita</i> seed oil was investigated using response surface methodology (RSM) to predict the optimal fat extraction conditions. The most efficient enzyme used in the experiment was a mixture of Flavourzyme<sup>®</sup> and Viscozyme<sup>®</sup> (1:1, <i>w</i>/<i>w</i>). The predicted maximum oil yield was 46.44%, using the following extraction conditions: 2.98% (<i>w</i>/<i>w</i>) enzyme loading, 48 °C incubation temperature and 76 min of incubation time. <i>H. heteroclita</i> seed oil obtained from heat and enzymatic pretreatments exhibited the highest lightness and viscosity. The chemical properties of this seed oil, including water and volatile compounds (≤0.2% <i>w</i>/<i>w</i>), acid value (≤4.0 mg KOH/g), peroxide value (≤15 mEq of active oxygen/kg) and soap content (≤0.005% <i>w</i>/<i>w</i>), were within the acceptable levels specified by the Codex Alimentarius (2019). <i>H. heteroclita</i> seed oil obtained from heat and enzymatic pretreatments contained the highest content of δ-tocopherol (88.29 mg/100 g) and omega-6 fatty acids (48.19 g/100 g). This study is the first to report on the enzyme-assisted mechanical extraction of <i>H. heteroclita</i> oilseeds as a promising plant material for vegetable oil production.
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