Studies On The Separation, Purification And Properties Of Fibrinolysis Enzyme In Lobster Sauce

Lobster Sauce is the traditional edible herbal medicine in China. Fibrinolysis enzyme is one kind of fibrinolysis active material contained in the lobster sauce.In this article, firstly, the fibrinolysis activities of 14 traditional lobster sauce were compared; next, representative lobster sauce of bacterial type was further studied. This paper explored the method for isolation and purification of fibrinolysis enzyme; established the relationship between proteolytic casein and fibrin plate method; properties and thrombolytic effect in vitro and vivo of fibrinolysis enzyme were researched by animal test. Primarily study result as follows:1. The fibrinolysis activities of 14 traditional lobster sauces were studied by fibrin plate method. The fibrinolysis activities were from 0.326 to 161.972 UK/mL. Mucor lobster sauce from Yongchuan showed the highest activity units in all samples, it reached to 161.972 UK/mL, aspergillus lobster sauce and peppery lobster sauce were lower.2.Separation and purification of fibrinolysis enzyme At first , precipitation with 75% ammonium sulfate to separate enzyme purification fold was 6.56 and recovery was 61.69%. Further, purification was achieved by DEAE-Sepharose FF, because this method was better than CM-Sepharose FF in the study, purification fold was 9.87 and recovery was 35.6%.Ultimately , fibrinolysis enzyme was purified up to 11.29 times with a recovery of 6.55% by Sephadex G-75.SDS-PAGE showed that no unpurposed protein was found after Sephadex G-75 chromatography, molecular weight was approximately 31KD.3. The properties of fibrinolysis enzyme Proteolytic casein method, a simpler and more effective method was established. The properties of fibrinolysis enzyme were determined by this method. The optimal reaction temperature and pH of fibrinolysis enzyme was 60℃, pH7～8 respectively .From pH 6～10, fibrinolysis enzyme exhibited high stability . Under 25℃and 37℃, after 4h the losing of activity was not visible. When temperature increased to 45℃, the enzyme activity decreased very fast, and...