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Purification and partial characterization of TFase, a fibrinolytic enzyme from the fruiting bodies of the medicinal and edible mushroom, Tremella fuciformis

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Abstract

A fibrinolytic metalloprotease-like enzyme named TFase, was purified from the fruiting bodies of the medicinal fungus Tremella fuciformis. Purification of TFase was performed by a chromatography on DEAE Sepharose CL-6B and Superdex-75. TFase showed a molecular weight of 38 kDa by SDS-PAGE and displayed a strong fibrin zymogram lysis band corresponding to a similar molecular mass. The enzyme demonstrated wide temperature optimum from 10 to 40°C and pH optimum at pH 8.0. The fibrinolytic activity of TFase was strongly inhibited by EDTA and EGTA, which are metalloprotease inhibitors. The K m and V max values for enzyme using the artificial substrate pyroGlu-Gly-Arg-pNA were 3.81 mM and 4.88 µmoles mg–1 min–1, respectively. TFase of T. fuciformis partially dissolved a fibrin clot directly and specifically cleaved the Aα and Bβ chains of fibrinogen. In addition, the enzyme prolonged the coagulation time, and increased the activated partial thromboplastin time and prothrombin time. These results suggest that TFase of T. fuciformis exhibits properties of a metalloprotease-like enzyme, shows novel fibrin(ogen)olytic enzyme activity, and has the potential to be a therapeutic agent for thrombosis.

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Authors and Affiliations

  1. Department of Biomedical Science, Chosun University, Gwangju, 501-759, Republic of Korea

    J.-H. Choi, D.-W. Kim, S.-E. Park & S.-J. Kim

  2. Department of Bio-Health Science, Gwangju University, Gwangju, 503-703, Republic of Korea

    S. Kim

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  1. J.-H. Choi
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  2. D.-W. Kim
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  3. S.-E. Park
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  4. S. Kim
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  5. S.-J. Kim
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Correspondence to S.-J. Kim.

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Choi, JH., Kim, DW., Park, SE. et al. Purification and partial characterization of TFase, a fibrinolytic enzyme from the fruiting bodies of the medicinal and edible mushroom, Tremella fuciformis . Appl Biochem Microbiol 51, 712–719 (2015). https://doi.org/10.1134/S0003683815060046

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  • DOI: https://doi.org/10.1134/S0003683815060046

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