NDLI: Purification and characterization of a fibrinolytic subtilisin-like protease of Bacillus subtilis TP-6 from an Indonesian fermented soybean, Tempeh

Content Provider	Springer Nature Link
Author	Kim, Seong Bo Lee, Dong Woo Cheigh, Chan Ick Choe, Eun Ah Lee, Sang Jae Hong, Young Ho Choi, Hak Jong Pyun, Yu Ryang
Copyright Year	2006
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 μM and 145 μmol 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 α- and β-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.
Starting Page	436
Ending Page	444
Page Count	9
File Format	PDF
ISSN	13675435
Journal	Journal of Industrial Microbiology and Biotechnology
Volume Number	33
Issue Number	6
e-ISSN	14765535
Language	English
Publisher	Springer-Verlag
Publisher Date	2006-02-10
Publisher Place	Berlin, Heidelberg
Access Restriction	Subscribed
Subject Keyword	Bacillus subtilis TP-6 Fibrin clot lysis Subtilisin-like protease Tempeh TPase
Content Type	Text
Resource Type	Article
Subject	Medicine Bioengineering Applied Microbiology and Biotechnology Biotechnology

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