Comparison of trypsin and chymotrypsin from the viscera of anchovy, Engraulis japonica.
The molecular weights of trypsin and chymotrypsin purified from anchovy viscera were estimated to be 25.6 and 26.1 Kda, respectively, by SDS-PAGE. Both enzymes had their maximal activity at pH 9.0 and 45 degrees C for casein and at pH 8.0 and 45 degrees C for synthetic substrates. Trypsin hydrolyzed at the position of Arg22 and Lys29, and chymotrypsin did at the position of Phe1, Tyr16, Phe24, Phe25, and Tyr26 of insulin beta-chain. The K'm and kcat of trypsin were 50 microM and 1.84 microM-1 min-1 toward N-benzoyl-L-arginine-p-nitroanilide (BAPNA) and those of chymotrypsin were 89 microM and 10.0 microM-1min-1 toward N-succinyl-(Ala)2-Pro-Phe-p-nitroanilide. The activation energy of trypsin and chymotrypsin were estimated to be 14 Kcal/mol toward N-benzoyl-L-arginine-p-nitroanilide and 6.5 Kcal/mol toward benzoyl-L-tyrosine ethyl ester.
Heu
M S
MS
Department of Nutrition and Food Science, National Fisheries University of Pusan, Korea.
Kim
H R
HR
Pyeun
J H
JH
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002918:Chymotrypsin / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004795:Enzyme Stability; D005399:Fishes / Q000378:metabolism*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011489:Protein Denaturation; D013379:Substrate Specificity; D013816:Thermodynamics; D014357:Trypsin / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D014361:Trypsin Inhibitors / Q000494:pharmacology; D014781:Viscera / Q000201:enzymology
D000596:Amino Acids; D014361:Trypsin Inhibitors; D002918:Chymotrypsin; D014357:Trypsin