Details

Autor(en) / Beteiligte

• Park, S.E.(Chosun Univ., Gwangju (Korea R.))
• Li, M.H
• Kim, J.S
• Sapkota, K
• Kim, J.E
• Choi, B.S
• Yoon, Y.H
• Lee, J.C
• Lee, H.H
• Kim, C.S
• Kim, S.J

Titel

Purification and characterization of a fibrinolytic protease from a culture supernatant of Flammulina velutipes mycelia

Ist Teil von

• Bioscience, biotechnology, and biochemistry, 2007-09, Vol.71 (9), p.2214-2222

Ort / Verlag

Tokyo: Japan Society for Bioscience, Biotechnology, and Agrochemistry

Erscheinungsjahr

2007

Quelle

MEDLINE

Beschreibungen/Notizen

• In this study we purified a fibrinolytic enzyme from the culture supernatant of Flammulina velutipes mycelia by ion exchange and gel filtration chromatographies, it was designated as F. velutipes protease (FVP-I). This purification protocol resulted in 18.52-fold purification of the enzyme at a final yield of 0.69%. The molecular mass of the purified enzyme was estimated to be 37 kDa by SDS-PAGE, fibrin-zymography and size exclusion by FPLC. This protease effectively hydrolyzed fibrin, preferentially digesting alpha-chain over beta-and gamma-gamma chain. Optimal protease activity was found to occur at a pH of 6.0 and a temperature of 20 to 30 deg C. The protease activity was inhibited by Cusup(2+), Fesup(2+) and Fesup(3+) ions, but was found to be enhanced by Mnsup(2+) and Mgsup(2+) ions. Furthermore, FVP-I activity was potently inhibited by EDTA and EGTA, and it was found to exhibit a higher specificity for chromogenic substrate S-2586 for chymotrypsin, indicating that the enzyme is a chymotrypsinlike metalloprotease. The first 20 amino acid residues of the N-terminal sequence of FVP-I were LTYRVIPITKQAVTEGTELL. They had a high degree of homology with hypothetical protein CC1G-11771, GeneBank Accession no. EAU86463.