'Journal of Experimental Biology and Agricultural Sciences'
Abstract
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
