CHARACTERIZATION OF ALPHA-AMYLASE FROM THE SEEDS OF Mucuna pruriens

Amylases are hydrolytic enzymes which are widely distributed in nature, animals, plants and microorganisms. Amylases are of great significance in present-day biotechnology. In present study, amylases are isolated from the soaked seeds of Mucuna pruriens under extreme acidic conditions. Conventional protein purification techniques such as salt fractionation, ion exchange chromatography on CM-cellulose and sephadex G-75 was employed for the purification of amylase from the seeds of Mucuna pruriens. The amylase activity was eluted in one peak. The specific activity and yield of the purified amylase was 6.25 and 29.99, respectively. Native PAGE, SDS-PAGE and gel electrofocussing were employed to establish homogeneity of the purified amylase. SDS-PAGE and gel-filtration chromatography on sephadex G-75 was used to determine the molecular weight of the purified amylase. The purified amylase was nearly homogenous and its molecular weight was found to be 78.4 kDa. The optimum pH and temperature of the purified amylase were 7.0 and 50oC, respectively. The isolectric pH of the purified amylase was 7.2 and the activity was linear up to 60 minutes

Anti-oxidant and anti-hyperglycemic properties of methanolic extracts of medicinal plants

Total phenolic content, DPPH free radical scavenging activity and alpha amylae inhibitory potential was determined for three selected medicinal plants - Gymnema sylvestre, Terminalia arjuna and Tinospora cordifolia. The plant extracts were prepared with methanol. The total phenolic content of methonolic extracts of Gymnema sylvestre, Terminalia arjuna and Tinospora cordifolia. were 6.862, 20.862 and 7.987 mg GAE/g plant material respectively . All the three plants showed anti oxidant activities with their IC50 values were 6.862, 20.862 and 7.987 μg/ml compared to IC50 value of the standard L-Ascorbic acid, which was 11.59μg/ml. The extracts of Gymnema sylvestre showed ±- amylase inhibition. Thus the results provided evidence that among the studied plants, Gymnema s ylvestre potential sources of natural antioxidant and antidiabetic activity