title: Production, Purification and Characterisation of a Potential Fibrinolytic Protease from Endophytic Xylaria curta by Solid Substrate Fermentation.
abstract: The present investigation highlights the optimal conditions for production of a non-toxic, bi-functional fibrinolytic enzyme xylarinase produced by endophytic fungus Xylaria curta by solid substrate fermentation using rice chaff medium. The purified enzyme is a monomeric protein with a molecular mass of ∼33 kDa. The enzyme exhibits cleavage of Aα and Bβ chains of fibrin(ogen) and has no effect on γ chain. The optimal fibrinolytic activity of the enzyme was observed at 35 °C and pH 8. The fibrinolytic activity was enhanced in the presence of Ca2+, whereas it was completely inhibited in the presence of Fe2+and Zn2+ions and inhibitors like EDTA and EGTA suggesting it to be a metalloprotease. The Kmand Vmaxof the enzyme for azocasein were 326 μM and 0.13 μM min-1. The N-terminal sequence of the enzyme (SNGPLPGGVVWAG) was same when compared to xylarinase isolated from culture broth of X. curta. Thus, xylarinase could be exploited as a potent clot busting enzyme which could be produced on large scale using solid substrate fermentation.
authors: Department of Biotechnology, Thapar University, Patiala, Punjab, 147004, India.
authors: Vineet
authors: V
authors: Meshram
authors: Department of Biotechnology, Thapar University, Patiala, Punjab, 147004, India. ssaxena@thapar.edu.
authors: Sanjai
authors: http://orcid.org/0000-0003-0025-8362
authors: S
authors: Saxena
authors: Department of Biochemistry, DAV University, Jalandhar, Punjab, India.
authors: Karan
authors: K
authors: Paul
authors: Department of Biotechnology, Thapar University, Patiala, Punjab, 147004, India.
authors: Mahiti
authors: M
authors: Gupta
authors: Department of Biotechnology, Thapar University, Patiala, Punjab, 147004, India.
authors: Neha
authors: N
authors: Kapoor
mesh_terms: D000818:Animals; D003470:Culture Media / Q000737:chemistry; D060026:Endophytes / Q000254:growth & development / Q000378:metabolism*; D005285:Fermentation; D005342:Fibrinolysis; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008667:Metalloproteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008670:Metals / Q000494:pharmacology; D051379:Mice; D008970:Molecular Weight; D012275:Oryza / Q000737:chemistry; D010802:Phylogeny; D011480:Protease Inhibitors / Q000494:pharmacology; D000067996:RAW 264.7 Cells; D013696:Temperature; D013098:Xylariales / Q000254:growth & development* / Q000378:metabolism*
chemical_list: D003470:Culture Media; D005656:Fungal Proteins; D008667:Metalloproteins; D008670:Metals; D011480:Protease Inhibitors; C000614656:xylarinase protein, Xylaria curta
keywords: Fibrinolysis; Metalloprotease; Solid-state fermentation; Thrombosis; Xylaria