Purification and characterization of a novel, highly potent fibrinolytic enzyme from Paecilomyces tenuipes

• Published in: Process biochemistry (1991) Vol. 46; no. 8; pp. 1545 - 1553
• Main Authors: Kim, Hoe Chang, Choi, Bong-Suk, Sapkota, Kumar, Kim, Seung, Lee, Hyo Jeong, Yoo, Jin Cheol, Kim, Sung-Jun
• Format: Journal Article
• Language: English
• Published: Elsevier Ltd 01-08-2011
• Subjects: calcium; entomopathogenic fungi; enzyme activity; fibrin; fibrinogen; Fibrinolysis; Fungal enzymes; humans; Paecilomyces; Paecilomyces tenuipes; polyacrylamide gel electrophoresis; serine proteinases; substrate specificity; Thrombosis; zinc; Paecilomyces tenuipes; Fungal enzymes; Fibrinolysis; Thrombosis
• ISSN: 1359-5113; 1873-3298
• DOI: 10.1016/j.procbio.2011.04.005

A fibrinolytic enzyme (PTEFP) was purified from the entomopathogenic fungus
Paecilomyces tenuipes. Analysis of the purified PTEFP by SDS–PAGE and fibrin zymography demonstrated a single protein band of approximately 14
kDa. Fibrinolysis pattern showed that PTEFP rapidly hydrolyzed α-chain followed by β-chain. PTEFP rapidly degraded Aα-chain of human fibrinogen but did not hydrolyze Bβ- or γ-chain indicating that it is α-fibrinogenase. The N-terminal sequence was AQNIGAVVNLSPPKQ, which is different from that of other known fibrinolytic enzymes. The PTEFP displayed maximum activity at 35
°C and pH 5.0, and was stable between pH 5.0–8.0 and below 40
°C. Calcium ion enhanced the enzyme activity whereas Zn
2+ inhibited it. The fibrinolytic activity was strongly inhibited by PMSF identifying it as a serine protease. PTEFP exhibited high specificity for the substrate H-D-Val-Leu-Lys-pNA and
K
m and
V
max values for this substrate were 0.17
mM and 59
U/ml respectively. These results suggest that PTEFP is a novel fibrinolytic enzyme and may have potential applications in treating thrombosis.