Fungal Source Of The Anticoagulant Activity Of Protein Tap

The blockage of the blood caused by thrombin is one of the most important factors thatcause people to die today. Therefore the research of new antithrombotic drugs is very valuable.The antithrombotic drugs can be divided into anticoagulant drugs, anti-platelet drugs andthrombolytic drugs. The metabolites from microorganisms are multiple and diverse. So they arethe abundant resources for screening antithrombotic agents.Fibrinogen can turn into fibrin by the catalysis of thrombin in the process of thrombosis.Basing on this catalytic reaction, we screened the fermentation solution of the actinomycetes,the mycelium extracts and the fermentation solution of some fungi. As a result, the fungiTermitomyces albuminosus CA03-1 had a stronger anticoagulant activity comparatively. So weselected this strain for further research work.We studied the most favorable isolation and purification of the anticoagulant agent fromTermitomyces albuminosus CA03-1. After the membranes filtration, HZ-816 absorb,ammonium sulphate precipitation, DEAE-Sephacel iron exchange and Sephadex G-100 gelfiltration, the anticoagulant protein(TAP) was isolated and purified. It appeared a single band onPAGE and two bands corresponding to substrates molecular weight of approximately 29.2kDand 21.9kD on SDS-PAGE. The TAP demonstrated a probably pI of 4.45 on IEF.The result indicated that the TAP was sensitive to temperature. The anticoagulant activity ofthe TAP was promoted by such salts as MgSO₄,CaCl₂,MnCl₂ and KCl. PMSF was a effectiveinhibitor of the TAP, which indicated the TAP might have a serine-related active center. Pepsinand trypsin tests showed that the TAP was tolerant to these two enzymes.