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Purification and characterization of fibrinolytic alkaline protease from Fusarium sp. BLB

  • Mitsuhiro Ueda
  • Toshihiro Kubo
  • Kazutaka Miyatake
  • Takumi Nakamura
Biotechnologically Relevant Enzymes and Proteins

Abstract

Fusarium sp. BLB, which produces a strongly fibrinolytic enzyme, was isolated from plant leaf (Hibiscus). Fibrinolytic alkaline protease was purified from a culture filtrate of Fusarium sp. BLB by precipitation with (NH4)2SO4 and column chromatography with CM-Toyopearl 650M and Superdex 75. The purified enzyme was homogeneous on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight was 27,000 by SDS-PAGE. Maximum activity of protease was observed at pH 9.5 and 50°C. Purified protease was active between pH 2.5 and 11.5 and was found to be stable up to 50°C. The enzyme derived from Fusarium sp. BLB is useful for thrombolytic therapy because this enzyme showed pH resistance. The activity was inhibited by diisopropylfluorophosphate and phenylmethylsulfonyl fluoride. The N-terminal amino acid sequence of the enzyme showed a similarity to those of proteases from Fusarium sp., Streptomyces griseus, Bos taurus bovine, Katsuwo pelamis digestive tract, and Lumbricus rubellus.

Keywords

Fibrinolytic activity Alkaline serine protease Fusarium sp 

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Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Mitsuhiro Ueda
    • 1
  • Toshihiro Kubo
    • 2
  • Kazutaka Miyatake
    • 1
  • Takumi Nakamura
    • 2
  1. 1.Laboratory of Biocycle Engineering, Graduate School of Life and Environmental SciencesOsaka Prefecture UniversitySakaiJapan
  2. 2.SODX CorporationIzumiJapan

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