Abstract: A hepatic stimulator substance from Shark liver (sHSS) was purified and characterized. The purification procedure included homogenization, centrifugation, ultrafiltration, DEAE-sephadex A25 chromatography and FPLC mono Q chromatography. By the procedure, the stimulatory specific activity of sHSS could be increased to 9000 IU/mg, and was 750 times more than the crude extract. The purity of the final purified fraction F70 which retained the maximal activity of sHSS was checked by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE) and High Performance Gel Filtration respectively, and showed only one band or one peak. Its molecular weight was measured to be 16973 Da by mass spectrometry. Its N-terminal sequence was determined to be N-Met-Arg-Thr-Gln-Glu-His-Thr-Lys- Ser-Val by PE-ABD 491A Protein N-Terminal Sequencing Meter. It was stable over a wide range of pH and temperature.

Key words: Hepatic stimulator substance, HSS, Shark, Purification, Mass spectrometry