A unique strong fibrinolytic enzyme (katsuwokinase) in skipjack "Shiokara," a Japanese traditional fermented food.
Katsuwokinase (KK) is a unique fibrinolytic enzyme recently found in skipjack "Shiokara," a Japanese traditional salt-fermented food. A crude enzyme extracted from skipjack Shiokara (Katsuwonus pelamis) showed a very strong fibrinolytic activity above 45 CU/g (fibrin plate method) based on plasmin. KK not only hydrolyzed fibrin but also several synthetic amido substrates, particularly pyro-Glu-Gly-Arg-pNA. The fibrinolytic activity of KK was not affected in the presence of 10% NaCl, was stable in the pH range from 1 to 10 at 37 degrees C for 30 min, and was inhibited by DFP, SBTI, BPTI, and aprotinin but not by epsilon-amino-n-caproic acid and t-4-amino-methylcyclohexane carboxylic acid. The crude enzyme contained at least four kinds of KK, and the major form purified had a pI value of approximately 5.0 and a molecular weight of 35,000. The N-terminal amino acid sequence of 21 residues, I-V-G-G-Y-E-Q-Z-A-H-S-Q-P-H-Q-V-S-L-N-S-G-, had 80% homology with that of trypsin. The fibrinolytic activity of the purified enzyme was approximately 2.6 times greater than that of plasmin by molar ratio, demonstrating its identity as a new and very potent fibrinolytic enzyme.
Sumi
H
H
Department of Nutrition, Faculty of Health and Welfare Science, Okayama Prefectural University, Japan.
Nakajima
N
N
Yatagai
C
C
D000595:Amino Acid Sequence; D010450:Endopeptidases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004795:Enzyme Stability; D005285:Fermentation; D005342:Fibrinolysis; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007564:Japan; D008969:Molecular Sequence Data; D017747:Seafood / Q000032:analysis*; D017385:Sequence Homology; D014357:Trypsin / Q000737:chemistry
D010450:Endopeptidases; D014357:Trypsin; C097306:katsuwokinase