29 kDa Trypsin from the pyloric ceca of Atlantic Bonito (Sarda sarda): recovery and characterization.
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.
Klomklao
Sappasith
S
Department of Food Science and Technology, Faculty of Technology and Community Development, Thaksin University, Phattalung Campus, Phattalung, 93110, Thailand.
Benjakul
Soottawat
S
Visessanguan
Wonnop
W
Kishimura
Hideki
H
Simpson
Benjamin K
BK
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
D014361:Trypsin Inhibitors; D014357:Trypsin