title: Purification and characterization of two trypsin-like enzymes from the digestive tract of anchovy Engraulis encrasicholus.
abstract: 1. Two trypsin-like enzymes, designated Trypsin A and B, were purified from the pyloric caeca and intestine of anchovy by (NH4)2SO4 fractionation, affinity chromatography (Benzamidine-Sepharose-6B) and ion exchange chromatography (DEAE-Sepharose). 2. Both trypsins catalyzed the hydrolysis of N-benzoyl-DL-arginine p-nitroanilide (BAPNA), p-tosyl-L-arginine methyl ester (TAME), casein and myofibrillar protein and they were inhibited by several well established trypsin-inhibitors. 3. The enzymes had mol. wts of 27,000 (Trypsin A) and 28,000 (Trypsin B). Their isoelectric points were about 4.9 (Trypsin A) and 4.6 (Trypsin B) and they had similar amino acid composition. 4. The enzymes had a pH optimum of 8-9 for the hydrolysis of BAPNA and of 9.5 for the digestion of casein and myofibrillar protein. Their activity and stability were affected by calcium ions. 5. Trypsins A and B resemble other fish trypsins in their mol. wt, pI, kinetic properties and the instability at low pH and they are similar to bovine trypsin in their dependence of Ca2+ for activity and stability.
authors.affiliation: Departamento de Bioquímíca y Biologia Molecular, Facultad de Ciencias, Universidad del País Vasco, Bilbao, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Martínez
authors.forename: R L
authors.initials: RL
authors.lastname: Olsen
authors.forename: J L
authors.initials: JL
authors.lastname: Serra
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000818:Animals; D004064:Digestive System / Q000201:enzymology*; D005399:Fishes / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007526:Isoelectric Point; D007700:Kinetics; D008970:Molecular Weight; D013379:Substrate Specificity; D013696:Temperature; D014357:Trypsin / Q000302:isolation & purification* / Q000378:metabolism; D014361:Trypsin Inhibitors / Q000494:pharmacology
chemical_list: D000596:Amino Acids; D014361:Trypsin Inhibitors; D014357:Trypsin