UFEIII, a fibrinolytic protease from the marine invertebrate, Urechis unicinctus

Abstract

A new serine protease with fibrinolytic activity from a marine invertebrate, Urechis unicinctus, was purified to electrophoretic homogeneity using column chromatography. SDS-PAGE of the purified enzyme showed a single polypeptide chain with MW ~20.8 kDa. Its N-terminal sequence was IIGGSQAAITSY. The purified enzyme, UFEIII, was stable at pH 6–10 below 60 °C with an optimum pH of 8.5 at approx. 55 °C. The enzyme activity was significantly inhibited by PMSF and SBTI suggesting that it was a serine protease. In fibrin plate assays, UFEIII was contained 1.46 × 10³ U (urokinase units) mg⁻¹ total fibrinolytic activity, which consisted of 692 U mg⁻¹ direct fibrinolytic activity and 769 U mg⁻¹ plasminogen-activator activity. K_m and V_max values for azocasein were 1 mg ml⁻¹ and 43 μg min⁻¹ ml⁻¹, respectively.