Steps for preparation

The Sesame Protein Isolate (SPI) was prepared from defatted sesame cake
Defatted sesame cake was milled using a home grinder.
Firstly, n-hexane was added at the ratio of 1:5 (w/v) with blending for 2 h and filtering to remove residual oil.
After that, the sesame cake was mixed with water at a ratio of 1:6 (w/v). The pH of the suspension was adjusted to pH 7.5 using 1.0 M NaOH, continuously stirred with a magnetic stirrer for 2 h and centrifuged at 5000 rpm for 15 min for removing of solid residues.
The supernatant containing soluble phases was collected and its pH was adjusted to pH 4.5 (pH isoelectric of SPI) using 0.1 M HCl due to which proteins precipitates.
The suspensions were centrifuged at 5000 rpm for 15 min, after which the supernatant was poured away. The precipitate obtained was washed twice, and re-suspended in a minimum amount of water. After neutralization to pH 7.0 with NaOH, the SPI sample was then freeze-dried and stored at 4 °C until analyzed and used.