Ann Genet Genet Disord | Volume 1, Issue 1 | Research Article | Open Access
Usharani Brammacharry1 and Muthuraj Muthaiah2*
1Department of Biomedical Genetics, University of Madras, India
2State TB Training and Demonstration Centre, Intermediate Reference Laboratory, India
*Correspondance to: Muthuraj Muthaiah
Fulltext PDFIn this study we endeavoured for the production, purification and characterization of anticoagulant property of metalloprotease enzyme from Pseudomonas fluorescens Migula B426 strain. Shake flask fermentation was performed to produce the enzyme and the fibrinolytic activities of the enzyme of this isolate. These were estimated to range across 40 to 5000 IU/mL of urokinase through the standard curve using the thrombolytic area on the fibrin plate. Purification of crude enzyme was done through Sephadex S-300 gel filtration and its activity was 2474 IU/mL.Enzyme activity was enhanced bydivalent cations Mg2+ and Ca2+ in the presence of Ethylene Diamine Tetra Acetic acid (EDTA), a metal-chelating agent and two metalloprotease inhibitors, 2, 2′-bipyridine and o-phenanthroline, repressed the enzymatic activity significantly. Amino acids of N-terminal sequence have great similarity with those of metalloprotease from various Pseudomonas strains and have consensus sequence, HEXXH zinc binding motif. These results strongly suggest that the extracellular protease enzyme of P.fluorescens Migula B426 strain is a novel zinc metalloprotease and is based on its N-terminal amino acid sequence, effect of protease inhibitors and its fibrinolytic activity. It can be further developed as a potential candidate for thrombolytic therapy.
Fibrin; Metalloprotease; Pseudomonas fluorescens; Zinc binding motif
Brammacharry U, Muthaiah M. Anticoagulant Property of Metalloprotease Produced by Pseudomonas fluorescens Migula B426. Ann Genet Genet Disord. 2018;1(1):1005.