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. 2006 Jun;33(6):436-44.
doi: 10.1007/s10295-006-0085-4. Epub 2006 Feb 10.

Purification and Characterization of a Fibrinolytic Subtilisin-Like Protease of Bacillus Subtilis TP-6 From an Indonesian Fermented Soybean, Tempeh

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Purification and Characterization of a Fibrinolytic Subtilisin-Like Protease of Bacillus Subtilis TP-6 From an Indonesian Fermented Soybean, Tempeh

Seong-Bo Kim et al. J Ind Microbiol Biotechnol. .

Abstract

We have isolated a bacterium (TP-6) from the Indonesian fermented soybean, Tempeh, which produces a strong fibrinolytic protease and was identified as Bacillus subtilis. The protease (TPase) was purified to homogeneity by ammonium sulfate fractionation and octyl sepharose and SP sepharose chromatography. The N-terminal amino acid sequence of the 27.5 kDa enzyme was determined, and the encoding gene was cloned and sequenced. The result demonstrates that TPase is a serine protease of the subtilisin family consisting of 275 amino acid residues in its mature form. Its apparent K (m) and V (max) for the synthetic substrate N-succinyl-Ala-Ala-Pro-Phe-pNA were 259 microM and 145 micromol mg(-1) min(-1), respectively. The fibrinogen degradation pattern generated by TPase as a function of time was similar to that obtained with plasmin. In addition, N-terminal amino acid sequence analysis of the fibrinogen degradation products demonstrated that TPase cleaves Glu (or Asp) near hydrophobic acids as a P1 site in the alpha- and beta-chains of fibrinogen to generate fragments D', E', and D' similar to those generated by plasmin. On plasminogen-rich fibrin plates, TPase did not seem to activate fibrin clot lysis. Moreover, the enzyme converted the active plasminogen activator inhibitor-1 to the latent form.

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References

    1. J Agric Food Chem. 2000 Aug;48(8):3210-6 - PubMed
    1. Nature. 1970 Aug 15;227(5259):680-5 - PubMed
    1. Biochem Biophys Res Commun. 1993 Dec 30;197(3):1340-7 - PubMed
    1. J Allergy Clin Immunol. 1985 Sep;76(3):453-7 - PubMed
    1. Blood. 1987 Jan;69(1):22-6 - PubMed

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