Kinetic Properties of Three Isoforms of Trypsin Isolated from the Pyloric Caeca of Chum Salmon (Oncorhynchus keta)(Analytical Biochemistry)
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概要
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Three isoforms of anionic chum salmon trypsin (ST-1, ST-2, and ST-3) were purified from the pyloric caeca of chum salmon (Oncorhynchus keta). The molecular weights of the three isoforms were about 24 kDa as determined by SDS-PAGE. The isoelectric points of ST-1, ST-2, and ST-3 were 5.8, 5.4, and 5.6, respectively. The apparent K_m values of two isoforms (ST-1 and ST-2) for BAPA (benzoyl-L-arginine-p-nitroanilide) hydrolysis at 5, 15, 25 and 35℃ were slightly higher than that of the main isoform ST-3, depending on temperature. The turnover numbers, k_<cat>, of ST-1 and ST-2 were about twice as high as that of ST-3. Consequently, the catalytic efficiencies (k_<cat>/k_m) of ST-1 and ST-2 were more efficient than ST-3. There were marked differences in both apparent K_m and k_<cat> values of three anionic chum salmon trypsins as compared to bovine cationic trypsin. K_m values of all chum salmon trypsins were approximately 10 times lower than those of bovine trypsin, depending on the temperature. The k_<cat> values of all chum salmon trypsins were about 2- to 5-fold higher than those of bovine trypsin; therefore, the catalytic efficiencies (k_<cat>/K_m) of chum salmon trypsin were 20- to 40-fold more efficient than those of bovine trypsin. On the other hand, k_<cat>/K_m values of ST-1 for TAME (tosyl-L-arginine methyl ester) hydrolysis were lower than those of bovine trypsin, whereas k_<cat>/K_m values of ST-2 and ST-3 were comparable to those of bovine trypsin, depending on the temperature.
- 2007-09-01
著者
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Toyota Eiko
Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido
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Sekizaki Haruo
Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido
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Iyaguchi Daisuke
Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido
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Iyaguchi Daisuke
Faculty Of Pharmaceutical Sciences Health Sciences University Of Hokkaido
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Toyota E
Faculty Of Pharmaceutical Sciences Health Sciences University Of Hokkaido
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Toyota Eiko
Faculty Of Pharmaceutical Sciences Health Sciences University Of Hokkaido
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Itoh Kunihiko
Faculty Of Pharmaceutical Sciences Health Sciences University Of Hokkaido
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Sekizaki H
Faculty Of Pharmaceutical Sciences Health Sciences University Of Hokkaido
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Sekizaki Haruo
Faculty Of Pharmaceutical Sciences Health Science University Of Hokkaido
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TANIZAWA Kazutaka
Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido
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Tanizawa K
Faculty Of Pharmaceutical Sciences Health Sciences University Of Hokkaido
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Sekizaki Haruo
Faculty of Pharmaceutical Science, Higashi Nippon Gakuen University
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Toyota Eiko
Faculty of Pharmaceutical Science, Higashi Nippon Gakuen University
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