Purification and characterization of a novel fibrinolytic enzyme from fruiting bodies of Korean Cordyceps militaris

• Published in: Bioresource technology 2011-02, Vol.102 (3), p.3279-3285
• Main Authors: Choi, DuBok, Cha, Wol-Suk, Park, Naomi, Kim, Hyun-Woo, Lee, Jong Hyuk, Park, Ji Seon, Park, Sang-Shin
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Biological and medical sciences; Chymotrypsin-like serine metalloprotease; Cordyceps - enzymology; Cordyceps militaris; Enzyme Activation; Enzyme Stability; Fibrin - chemistry; Fibrinogenolysis; Fibrinolysis; Fibrinolytic enzyme; Fruit; Fundamental and applied biological sciences. Psychology; Molecular Sequence Data; Peptide Hydrolases - chemistry; Peptide Hydrolases - isolation & purification
• ISSN: 0960-8524; 1873-2976
• DOI: 10.1016/j.biortech.2010.10.002

A fibrinolytic enzyme has been purified from the fruiting bodies of Korean Cordyceps militaris. The molecular mass of the enzyme was estimated to be 34kDa by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE), fibrin-zymography, and gel filtration chromatography. The 15 amino acid residues of the N-terminal sequence of the enzyme were APVEQCDAPVGLARL, which is dissimilar to those of fibrinolytic enzymes from other mushrooms. Optimal pH and temperature values of the enzyme were 7.0 and 40°C, respectively. The enzyme activity was completely inhibited by phenylmethylsulfonyl fluoride (PMSF), TPCK, 1,10-phenanthroline, Cu2+, and Ba2+. It was also significantly inhibited by aprotinin, EDTA, and EGTA. The enzyme showed a higher specificity for a synthetic substrate, N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide, exhibiting that it is a chymotrypsin-like serine metalloprotease. The enzyme preferentially hydrolyzed the fibrinogen Aα-, followed by the Bβ-chains and the γ-chain. The α, β, and γ–γ chains of fibrin were also degraded by the enzyme.