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Plant endophytes are among the most important resources of biologically active metabolites.Twenty-three endophytestrains residing in Trachelospermum jasminoides were cultivated in vitro with the cultures assayed for the fibrinolyticsubstance production.As a result,the culture of Verticillium sp.Tj33 was shown to be the most active.A fibrinolytic enzymedesignated as verticase was subsequently purified from the supernatant of Verticillium sp.culture broth by a combinationof DEAE-52,Sephadex G-75 and hydrophobic column chromatographies.Verticase,with its molecular mass of 31 kDaand pl of 8.5,was demonstrated to be homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis andisoelectric focusing electrophoresis.Verticase is an enzyme that hydrolyzes fibrin directly without activation of plaminogen.It was stable in a broad pH range from 4 through to 11 with the optimal reaction pH value and temperature shown to bearound 9-10 and 50-60℃,respectively.The fibrinolytic activity of verticase was severely inhibited by phenylmethylsulfonyfluoride,indicating that verticase was a serine protease.
Plant endophytes are among the most important resources of biologically active metabolites. Twenty-three endophytestrains residing in Trachelospermum jasminoides were cultivated in vitro with the cultures assayed for the fibrinolytic assay production. As a result, the culture of Verticillium sp. Tj33 was shown to be the most active. A fibrinolytic enzyme designated as verticase was purified from the supernatant of Verticillium sp. culture broth by a combination of DEAE-52, Sephadex G-75 and hydrophobic column chromatographies. Vernase, with its molecular mass of 31 kDa and pl of 8.5, was demonstrated to be homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis andisomeric focusing electrophoresis. Veertase is an enzyme that hydrolyzes fibrin directly without activation of plaminogen. It was stable in a broad pH range from 4 through to 11 with the optimal reaction pH value and temperature shown to bearound 9-10 and 50-60 ° C, respectively.The fibrinolytic activity of verticase was severely inhibited by phenylmethylsulfonyfluoride, indicating that verticase was a serine protease.