Purification and characterization of a novel fibrinolytic enzyme from Whitmania pigra Whitman.
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+, Mg2+,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.
a Marine College, Shandong University , Weihai , China.|b Weihai International Biotechnology Research and Development Center, Shandong University , Weihai , China.
Fulong
F
Chu
a Marine College, Shandong University , Weihai , China.|b Weihai International Biotechnology Research and Development Center, Shandong University , Weihai , China.
Xiaochen
X
Wang
a Marine College, Shandong University , Weihai , China.|b Weihai International Biotechnology Research and Development Center, Shandong University , Weihai , China.
Qianqian
Q
Sun
a Marine College, Shandong University , Weihai , China.|b Weihai International Biotechnology Research and Development Center, Shandong University , Weihai , China.
Hao
H
Liang
d Zaozhuang Mining Group Central Hospital , Zaozhuang , China.
Sijian
S
Wang
a Marine College, Shandong University , Weihai , China.|b Weihai International Biotechnology Research and Development Center, Shandong University , Weihai , China.
Dengkun
D
An
a Marine College, Shandong University , Weihai , China.|b Weihai International Biotechnology Research and Development Center, Shandong University , Weihai , China.
Chao
C
Cui
a Marine College, Shandong University , Weihai , China.|b Weihai International Biotechnology Research and Development Center, Shandong University , Weihai , China.
Yuchao
Y
Chai
a Marine College, Shandong University , Weihai , China.|b Weihai International Biotechnology Research and Development Center, Shandong University , Weihai , China.
Shuaishuai
S
Li
a Marine College, Shandong University , Weihai , China.|b Weihai International Biotechnology Research and Development Center, Shandong University , Weihai , China.
Shuliang
S
Song
a Marine College, Shandong University , Weihai , China.|b Weihai International Biotechnology Research and Development Center, Shandong University , Weihai , China.|c School of Pharmaceutical Sciences, Shandong University , Jinan , China.
Aiguo
A
Ji
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D057075:Enzyme Assays / Q000379:methods; D004798:Enzymes / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D005343:Fibrinolytic Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007865:Leeches / Q000201:enzymology*; D008970:Molecular Weight; D013379:Substrate Specificity; D013696:Temperature
D004798:Enzymes; D005343:Fibrinolytic Agents
Fibrinolytic enzyme; Whitmania pigra Whitman; fibrinogenolytic; fibrinolytic; purification