title: Isolation of soybean trypsin inhibitors by affinity chromatography on anhydrotrypsin-Sepharose 4B.
abstract: By repeated treatments of trypsin with phenylmethylsulfonyl fluoride (PMSF), followed by base elimination of PMS from the PMS-trypsin, a catalytically inactive anhydrotrypsin preparation of low (less than 1%) active trypsin content was obtained. Inactive material was removed by affinity chromatography on trypsin inhibitor-Sepharose 4B and the purified anhydrotrypsin with full binding capacity for trypsin inhibitors was coupled to cyanogen bromide-activated Sepharose 4B. When used below its maximum capacity for trypsin inhibitors the anhydrotrypsin-Sepharose-4B affinity column absorbed both classes of inhibitors present in soybean. When overloaded, the Kunitz type was bound preferentially. Based on this observation, conditions for the partial separation of the two types of inhibitors were worked out.
authors.affiliation: Rowett Research Institute, Bucksburn, Aberdeen, Scotland, United Kingdom.
authors.forename: A
authors.initials: A
authors.lastname: Pusztai
authors.forename: G
authors.initials: G
authors.lastname: Grant
authors.forename: J C
authors.initials: JC
authors.lastname: Stewart
authors.forename: W B
authors.initials: WB
authors.lastname: Watt
mesh_terms: D002846:Chromatography, Affinity; D004589:Electrophoresis, Disc; D007525:Isoelectric Focusing; D012685:Sepharose; D013025:Glycine max / Q000032:analysis; D014357:Trypsin / Q000138:chemical synthesis; D014361:Trypsin Inhibitors / Q000302:isolation & purification*
chemical_list: D014361:Trypsin Inhibitors; C011661:anhydrotrypsin; D012685:Sepharose; D014357:Trypsin