Khan Sabir, Arshad Shafia, Arif Amina, Tanveer Rida, Amin Zeemal Seemab, Abbas Saba, Maqsood Amna, Raza Muhammad, Munir Arooj, Latif Amara, Habiba Maryam, Afzal Muhammad
Tti Testing Laboratories, Lahore, Pakistan.
UCCM, Faculty of Medicine and Allied Health Sciences, Islamia University Bahawalpur, Punjab , Pakistan.
Dose Response. 2022 Oct 12;20(4):15593258221131462. doi: 10.1177/15593258221131462. eCollection 2022 Oct-Dec.
The current study aims to isolate, purify, and characterize the trypsin inhibitor protein from seeds of soya beans, scientifically known as . 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. 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.
当前的研究旨在从大豆种子中分离、纯化并鉴定胰蛋白酶抑制剂蛋白,大豆种子的学名为 。将其种子研磨,然后用正己烷浸泡粉末数次。将其加入磷酸盐缓冲盐水(PBS),随后对溶解有PBS的提取物进行过滤和离心。将上清液进行硫酸铵沉淀,并制备约六个级分,即30%至80%的级分。从每个级分中获得的离心沉淀进行透析,并在SDS-PAGE上进行电泳。胰蛋白酶抑制剂蛋白在30%的沉淀中沉淀并得到鉴定,与蛋白标准品(赛默飞世尔科技10 - 170 kDa)相比,其分子量为21.5 kDa。气相色谱-质谱联用(GC-MS)分析揭示了诸如豆甾醇、菜油甾醇、β-谷甾醇和γ-生育酚等类固醇衍生物。胰蛋白酶抑制剂可作为一种植物源药物,用于在没有真正底物(蛋白质)被激活并开始自消化导致胰腺炎的情况下,克服胰蛋白酶的过度激活。