Trypsin Inhibitor Isolated From Glycine max (Soya Bean) Extraction, Purification, and Characterization

Abstract

The current study aims to isolate, purify, and characterize the trypsin inhibitor protein from seeds of soya beans, scientifically known as Glycine max. Its seeds were ground, and the powder was soaked several times using n-hexane. It was added to phosphate buffer saline (PBS) followed by filtration and centrifugation of the PBS dissolved extract. The supernatant was subjected to ammonium sulfate precipitation and about six fractions, 30% to 80% were prepared. The centrifuged pellets obtained from each fraction were dialyzed and run on SDS-PAGE. The trypsin inhibitor protein was precipitated and characterized in 30% pellet and molecular weight was 21.5 kDa compared to protein ladder (ThermoFisher 10-170 kDa). GC-MS analysis revealed the steroid derivatives such as stigmasterol, campesterol, beta-sitosterol, and gamma-tocopherol. Glycine max trypsin inhibitor could be used as a plant-derived drug to overcome the over-activation of trypsin without its real substrate (proteins) becoming activated and start auto digestion leading to pancreatitis.

Advancing Faba Bean Protein Purification Using Membrane Technology: Current State and Future Perspectives

“…It was found that ultrafiltration did not completely remove salt from the Membrane dialysis has also been used in protein purification to improve separation efficiency and yield. In a study on protein extraction from soybean, Khan et al [103] used membrane dialysis to remove the salt from the trypsin inhibitor protein extract. The supernatant from protein extraction was subjected to centrifugation followed by ammonium sulfate precipitation.…”