title: Alcalase rapeseed inhibitors: purification and partial characterization.
abstract: Extensive rapeseed protein hydrolysate obtained sequentially with Alcalase and Flavourzyme showed inhibitory activity towards Alcalase. Inhibitory activity decreased as the hydrolytic process progressed probably by heat denaturation and/or partial protease degradation. Alcalase rapeseed inhibitors were purified by gel filtration and subsequent ion exchange chromatography. They are composed of peptides of 8.4 and 6.1 kDa linked by interchain disulphide bonds, as observed by reducing SDS-PAGE, with a native molecular weight of 18 kDa. Aminoacid composition of the inhibitors was characterized by the high proportion of methionine (4.2%) and cysteine (4.6%). Alcalase inhibitors were partially resistant to heat treatment; after heating at 70 degrees C for 45 minutes more than 50% of the original inhibitory activity remained in the purified protein but after heating at 90 degrees C for 5 minutes, inhibitory activity decreased very fast to a basal level. The possible relation of these protease inhibitors with the 2S albumin storage proteins is discussed.
authors.affiliation: Instituto de la Grasa, Sevilla, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Vioque
authors.forename: R
authors.initials: R
authors.lastname: Sánchez-Vioque
authors.forename: A
authors.initials: A
authors.lastname: Clemente
authors.forename: J
authors.initials: J
authors.lastname: Pedroche
authors.forename: M
authors.initials: M
authors.lastname: Mar Yust
authors.forename: F
authors.initials: F
authors.lastname: Millán
mesh_terms: D000596:Amino Acids / Q000032:analysis; D029741:Brassica rapa / Q000737:chemistry*; D006358:Hot Temperature; D006868:Hydrolysis; D010940:Plant Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D011480:Protease Inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D011489:Protein Denaturation; D013381:Subtilisins / Q000037:antagonists & inhibitors*
chemical_list: D000596:Amino Acids; D010940:Plant Proteins; D011480:Protease Inhibitors; D013381:Subtilisins