Inhibitors of human and bovine trypsin and chymotrypsin in fenugreek (Trigonella foenum-graecum L.) seeds. Demonstration and purification.
Fenugreek contained proteinase inhibitors inhibiting 5-9 mg human trypsin, 5-7 mg bovine trypsin, 2-6 mg human chymotrypsin, and 1-3 mg bovine chymotrypsin per g seed material. About 30 inhibitors were electrophoretically detected, and 23 of them, inhibiting all the four enzymes, were characterized by means of their isoelectric points: a group of acid inhibitors (TFI-A1 to A10, pI 4.48-5.12), a group of neutral inhibitors (TFI-N1 to -N6, pI 5.91-6.71), and a group of basic inhibitors (TFI-B1 to -B7, pI 7.76-9.77). To eliminate the galactomannans which complicate further purification, coarsely ground seeds were separated by density into two fractions, seed coats + endosperm and cotyledons + embryos (C + E). Isolation of the fenugreek inhibitors by extraction of fraction C + E, followed by ammonium sulfate fractionation and affinity chromatography on anhydrotrypsin-Sepharose, resulted in an about 700-fold enrichment.
Weder
J K
JK
Institut für Lebensmittelchemie, Technische Universität München, Garching, Federal Republic of Germany.
Haussner
K
K
D000818:Animals; D002417:Cattle; D002918:Chymotrypsin / Q000037:antagonists & inhibitors*; D007887:Fabaceae / Q000032:analysis*; D006801:Humans; D007526:Isoelectric Point; D010946:Plants, Medicinal; D012639:Seeds / Q000032:analysis*; D014361:Trypsin Inhibitors / Q000737:chemistry / Q000302:isolation & purification*
D014361:Trypsin Inhibitors; D002918:Chymotrypsin