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Purification and characterization of a novel fibrinolytic protease from Fusarium sp. CPCC 480097

  • Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

A novel fibrinolytic enzyme from Fusarium sp. CPCC 480097, named Fu-P, was purified to electrophoretic homogeneity using ammonium sulfate precipitation and ion exchange and gel filtration chromatography. Fu-P, a single protein had a molecular weight of 28 kDa, which was determined by SDS-PAGE and gel filtration chromatography. The isoelectric point of Fu-P determined by isoelectric focusing electrophoresis (IEF) was 8.1, and the optimum temperature and pH value were 45°C and 8.5, respectively. Fu-P cleaved the α-chain of fibrin (ogen) with high efficiency, and the β-chain and γ-γ (γ-)-chain with lower efficiency. Fu-P activity was inhibited by EDTA and PMSF, and the enzyme exhibited a high specificity for the chymotrypsin substrate S-2586. Fu-P was therefore identified as a chymotrypsin-like serine metalloprotease. The first 15 amino acids of the N-terminal sequence of Fu-P were Q-A-S–S-G-T-P-A-T-I-R-V-L-V–V and showed no homology with that of other known fibrinolytic enzymes. This protease may have potential applications in thrombolytic therapy and in thrombosis prevention.

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Acknowledgments

This work was support by the National Infrastructure of National Resources for Science and Technology (grant no 2005DKA21203). Isoelectric point and the N-terminal amino acid sequence of Fu-P were measured by Shanghai Institutes for Biological Sciences (Chinese Academy of Sciences).

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Authors and Affiliations

  1. Department of Biological Pharmacy, Shanghai Institute of Pharmaceutical Industry, 200040, Shanghai, China

    Bin Wu & Daijie Chen

  2. Department of Life Science and Biopharmacy, Shenyang Pharmaceutical University, 110016, Shenyang, China

    Licheng Wu

  3. Shanghai Health Creation Center of Biopharmaceutical R&D, 201203, Shanghai, China

    Zhijun Yang & Minyu Luo

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  1. Bin Wu
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  2. Licheng Wu
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  3. Daijie Chen
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  4. Zhijun Yang
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  5. Minyu Luo
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Correspondence to Minyu Luo.

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Wu, B., Wu, L., Chen, D. et al. Purification and characterization of a novel fibrinolytic protease from Fusarium sp. CPCC 480097. J Ind Microbiol Biotechnol 36, 451–459 (2009). https://doi.org/10.1007/s10295-008-0516-5

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  • DOI: https://doi.org/10.1007/s10295-008-0516-5

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