Isolation and Characterization of a Fibrinolytic Enzyme from Cultured Sipunculid Phasclosoma esculenta
CAI Binxin1,2, ZHOU Fengfang1,2,3, RUAN Shaojiang1,2, HUANG Weiqing1,2,3, LIU Congying1, SU Yu1
1. College of Life Science, Ningde Normal University, Ningde 352100, China; 2. State Oceanic Administration Hercynian Special Biological Germplasm Resources and Biological Product Development Public Service Platform, Ningde 352100, China; 3. Engineering Research Center of Mindong Aquatic Product Deep-Processing of Fujian Universities, Ningde 352100, China
Abstract:The fibrinolytic enzyme was extracted from coelomic fluid, intestine, esophagus of cultured sipunculid Phascolosoma esculenta by precipitation, Q sepharose fast flow anion-exchange and sephcry1 S-100 gel filtration, and located in the intestine. Effects of pH (3, 4, 5, 6, 7, 8, 9, and 10), temperature (4 ℃, 37 ℃, 50 ℃, 60 ℃, and 70 ℃), metal ions (Na+, K, Ca2+, Mg2+, Fe2+, Cu2+, and Al3+) on enzyme activity and ability to dissolve blood clots in vitro were investigated in the fibrinolytic enzyme of sipunculid. The results showed that a fibrinolytic enzyme extracted from intestine of sipunculid known as PK had strong ability to dissolve fibrinolytic protein, with relative molecular mass of about 32 ku, and specific activity of 4343.12 U/mg, increased by 13.82 folds compared with that in crude extract enzyme. But only 3.83% of total enzyme activity recovery was observed. The stable activity of the PK, with blood clot solubility of 88.09%, was kept at pH from 6 to 9 and below 37 ℃, and activated by Mg2+ (P<0.05) and inhibited by Fe2+, Cu2+, and Al3+ (P<0.05). The findings indicated that there was fibrinolytic enzyme activity in the intestine of sipunculid, which provided reference with development of fibrinolytic enzyme.
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