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

Bioresour Technol. 2011 Feb;102(3):3279-85. doi: 10.1016/j.biortech.2010.10.002. Epub 2010 Oct 16.

Abstract

A fibrinolytic enzyme has been purified from the fruiting bodies of Korean Cordyceps militaris. The molecular mass of the enzyme was estimated to be 34 kDa 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, Cu(2+), and Ba(2+). 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Cordyceps / enzymology*
  • Enzyme Activation
  • Enzyme Stability
  • Fibrin / chemistry*
  • Fruit
  • Molecular Sequence Data
  • Peptide Hydrolases / chemistry*
  • Peptide Hydrolases / isolation & purification*

Substances

  • semen liquefaction factor
  • Fibrin
  • Peptide Hydrolases