title: 29 kDa Trypsin from the pyloric ceca of Atlantic Bonito (Sarda sarda): recovery and characterization.
abstract: Trypsin from the pyloric ceca of Atlantic bonito (Sarda sarda) was purified and characterized with respect to its purity; molecular weight; sensitivity to temperature, pH, and inhibition; and N-terminal sequence. The purified trypsin had a molecular weight of 29 kDa as per sodium dodecyl sulfate polyacrylamide gel electrophoresis, and optimal activity was observed at pH 9 and 65 degrees C with BAPNA as a substrate. The enzyme was stable to heat treatment up to 50 degrees C and within the pH range of 7-12. It was stabilized by calcium ions, but its activity was strongly inhibited by soybean trypsin inhibitor, N-p-tosyl-L-lysine chloromethyl ketone, and phenyl methyl sulfonyl fluoride. The enzyme exhibited a progressive decrease in activity with increasing NaCl concentration (0-30%). The N-terminal 20 amino acid residues of Atlantic bonito trypsin were determined as IVGGYECQAHSQPWQPVLNS and were homologous with other trypsins.
authors.affiliation: Department of Food Science and Technology, Faculty of Technology and Community Development, Thaksin University, Phattalung Campus, Phattalung, 93110, Thailand.
authors.forename: Sappasith
authors.initials: S
authors.lastname: Klomklao
authors.forename: Soottawat
authors.initials: S
authors.lastname: Benjakul
authors.forename: Wonnop
authors.initials: W
authors.lastname: Visessanguan
authors.forename: Hideki
authors.initials: H
authors.lastname: Kishimura
authors.forename: Benjamin K
authors.initials: BK
authors.lastname: Simpson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002432:Cecum / Q000201:enzymology*; D004795:Enzyme Stability; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010473:Perciformes / Q000378:metabolism*; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein; D014357:Trypsin / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D014361:Trypsin Inhibitors / Q000494:pharmacology
chemical_list: D014361:Trypsin Inhibitors; D014357:Trypsin