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Purification and characterization of fibrinolytic protease from Bacillus amyloliquefaciens MCC2606 and analysis of fibrin degradation product by MS/MS.

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Relation http://ir.cftri.com/13591/
https://doi.org/10.1080/10826068.2017.1421964
 
Title Purification and characterization of fibrinolytic protease from Bacillus
amyloliquefaciens MCC2606 and analysis of fibrin degradation product by MS/MS.
 
Creator Yogesh, Devaraj.
Savita Kumari, R.
Prakash, M. Halami
 
Subject 03 Proteins
02 Fermented foods
 
Description A serine protease with preference for fibrin protein was purified and characterized from Bacillus
amyloliquefaciens MCC2606, isolated from dosa batter. The protease was purified using ammonium
sulfate precipitation, ion-exchange, and gel filtration chromatography. The degradation activity of the
protease toward the fibrin was significantly higher compared with other protein substrates in the study.
The molecular weight of the CFR15-protease was estimated to be 32 kDa based on SDS-PAGE. The
purified enzyme exhibited both fibrinolytic and fibrinogenolytic activity. The optimum pH and
temperature for the activity of the enzyme was found to be 10.5 and 45°C. A significant inhibition was
seen with the protease inhibitors phenyl methyl sulphonyl fluoride and ethylene diamine tetra acetic acid
and the activity of the enzyme was enhanced in presence of Mn2+. There was an observed increase in
vitro activated partial thromboplastin time and prothrombin time of both time and dose dependent
study. Among the four chains of fibrin, the β-chain of fibrin appears to be the primary component and
site susceptible for CFR15-protease in early action as indicated by MS/MS analysis of initial degradation
products. These results indicated that the CFR15-protease have the potential to be an effective
fibrinolytic agent.
 
Publisher Taylor & Francis
 
Date 2018
 
Type Article
PeerReviewed
 
Format pdf
 
Language en
 
Identifier http://ir.cftri.com/13591/1/PREPARATIVE%20BIOCHEMISTRY%20AND%20BIOTECHNOLOGY%202018%2C%20VOL.%2048%2C%20NO.%202%2C%20172%E2%80%93180.pdf
Yogesh, Devaraj. and Savita Kumari, R. and Prakash, M. Halami (2018) Purification and characterization of fibrinolytic protease from Bacillus amyloliquefaciens MCC2606 and analysis of fibrin degradation product by MS/MS. Preparative Biochemistry and Biotechnology, 48 (2). pp. 172-180. ISSN 1082-6068