open access

https://hdl.handle.net/2115/49331

Purification and characteristics of trypsin from masu salmon (Oncorhynchus masou) cultured in fresh-water

英語

Phylogenetic tree

2010-09

Springer Netherlands

Fish Physiology and Biochemistry

36(3)

637-645

Trypsin from the pyloric ceca of masu salmon (Oncorhynchus masou) cultured in fresh-water was purified by a series of chromatographies including Sephacryl S-200, Sephadex G-50 and diethylaminoethyl-cellulose to obtain a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and native-PAGE. The molecular mass of the purified trypsin was estimated to be approximately 24,000 Da by SDS-PAGE. The enzyme activity was strongly inhibited by phenylmethylsulfonyl fluoride, soybean trypsin inhibitor, and Nα-p-tosyl-L-lysine chloromethyl ketone. Masu salmon trypsin was stabilized by calcium ion. The optimum pH of the masu salmon trypsin was around pH 8.5 and the trypsin was unstable below pH 5.0. The optimum temperature of the masu salmon trypsin was around 60℃, and the trypsin was stable below 50℃, like temperate-zone and tropical-zone fish trypsins. The N-terminal 20 amino acid sequence of the masu salmon trypsin was IVGGYECKAYSQPHQVSLNS, and its charged amino acid content was lower than those of trypsins from frigid-zone fish and similar to those of trypsins from temperate-zone and tropical-zone fish. In phylogenetic tree, the masu salmon trypsin was classified into the group of the temperate-zone fish trypsin.

学術雑誌論文

Accepted Manuscript

The final publication is available at www.springerlink.com

0920-1742

1573-5168