Pharmacotherapy Group, Faculty of Pharmacy, University of Benin, Benin City, Nigeria
Abstract
Purpose: To isolate, purify and characterize protease from the latex of
the plant. Methods: Protease was isolated from the latex of Plumeria
rubra Linn using acetone precipitation method and purified by a
sequence of DEAE cellulose column chromatography, followed by two
successive column purification in Sephadex G-50 and Sephadex G-200. The
molecular weight of the purified protease was determined by sodium
dodecyl sulfate polyacrylamide gel electrophoresis (SDSPAGE). The
protease was given a trivial name, Plumerin-R. Results: Plumerin-R
showed a single protein band on SDS-PAGE and molecular weight was
approximately 81.85 kDa. It remained active over a broad range of
temperature but had optimum activity at 55 °C and pH 7.0 when
casein was used as substrate. Activation of the protease by a
thiol-activating agent indicated the presence of sulfhydryl as an
essential group for its activity. Conclusion: A protease from the latex
of Plumeria rubra Linn was purified to homogeneity by a simple
purification procedure and then characterized
