ABSTRACT

Purification and characterization of MCase from Momordica Charantia Seed Extract was carried by fractionation, on DEAE-Cellulose column chromatography and sephadex G-75 column chromatography. When peak II was fractionated on anion exchange, four peaks were resolved (Peak I-IV), but only peak II exhibited strong anticoagulant activity. In addition, peak II showed sharp single band on SDS-PAGE with an apparent molecular weight of 100kDa in both non- reducing and reducing conditions. Purity of that protein was adjudged using RP-HPLC. The purified proteins showed strong proteolytic activity with the specific activity of 14.84 units/mg/min. Protease inhibition studies were performed using specific inhibitors such as PMSF, IAA, EDTA and 1,10-Phenanthroline. Interestingly, protease was susceptible to PMSF but insensitive to remaining, suggesting the purified protein is of serine family. Hence, it named as MCase. MCase hydrolysed fibrinogen and fibrin as well. The optimum activity of serine proteases was observed in the temperature range 37-40 0 C and PH between 8-8.5. MCase found to exhibit anticoagulant activity was confirmed by plasma recalcification time and mouse tail bleeding assay. Keywords: MCase, Momordica Charantia, SDS-PAGE, RP-HPLC
Publication date: 01/05/2023

https://ijbpas.com/pdf/2023/May/MS_IJBPAS_2023_7134.pdf

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https://doi.org/10.31032/IJBPAS/2023/12.5.7134