Purification and characterization of a novel fibrinolytic enzyme from Whitmania pigra Whitman

ABSTRACT

A fibrinolytic enzyme was purified from the dry body of Whitmania pigra Whitman. The fibrinolytic enzyme was purified to homogeneity with a yield of 0.003% and a purification of 630.7 fold. The molecular weight of the enzyme was estimated to be 26.7 kDa by reduced sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was tested by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and it showed that the enzyme was a novel fibrinolytic enzyme. The optimal pH and temperature of the enzyme were 8.5 and 55°C, respectively. Enzyme activity was enhanced by Na⁺, Mg^2+, and K⁺. On the contrary, the proteolytic activity was significantly inhibited by Mn²⁺, Fe²⁺, Fe³⁺, ethylenediaminetetraacetic acid (EDTA), and ethylenebis(oxyethylenenitrilo)tetraacetic acid (EGTA). Fibrinolytic and fibrinogenolytic assays showed that the enzyme preferentially hydrolyzed fibrinogen Aα-chains, followed by Bβ- and γ-chains. The α-, β-, and γ-γ-chains of fibrin were also degraded by the enzyme.

KEYWORDS:

• Fibrinolytic enzyme
• fibrinolytic
• fibrinogenolytic
• purification
• Whitmania pigra Whitman

Declaration of interest

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.

Funding

This work was supported by the Shandong Provincial Natural Science Foundation, China (ZR2015PH050).

Additional information

Funding

This work was supported by the Shandong Provincial Natural Science Foundation, China (ZR2015PH050).