Purification and Characterization of a-Galactosidase from Artocarpus Hirsutus Seeds

This paper describes that the work done on the isolation and purification of a-galactosidase from Artocarpus hirsutus seeds has been purified using a combination of various fractionation and chromatographic techniques following ion-exchange chromatography CM-cellulose and gel filtration on Superdex G- 100, giving a final specific activity of 0.175 μmoles/min/mg with a fold purification of 6.48 and yield of 1.29%. Analysis of the protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed two low molecular weight protein bands. The optimum pH of the enzyme was determined. The enzyme was stable over a pH range of 3.0-7.0 and a temperature range of 7-45 degree C. Sequence alignment of the A. hirsutus seed a- galactosidase enzyme with other known plant a-galactosidase showed high amino acid sequence homology. . KEYWORDS :Artocarpus hirsutus, Galactosidases, Gel-permeation, Ion exchange, Purification, Sodium dodecyl sulfate-polyacrylamide gel electrophoresis