Purification and characterization of alpha-amylase from apple (Malus pumila)
2004
Kanwal, B. (Government Coll., Sahiwal (Pakistan). Dept. of Botany) | Zia, M.A. | Yasin, M. | Rahman, K. | Sheikh, M.A.
Malus pumila (apple) was homogenized in buffer for a-amylase extraction. The crude extract showed 3.09 U/mL enzyme activity that was subjected to ammonium sulfate precipitation and 4.76 U/mL of activity was obtained with 5.01 U/mg specific activity. It was applied to sephadex G-150 for gel filtration chromatography. It indicated the activity of 5.025 U/mL and specific activity 38.95 U/mL with 20-fold purification. SDS-PAGE of enzyme showed that unwanted proteins are removed and clear band of enzyme was appeared. Molecular weight was determined by sephadex G-150 column that is 51,180 Dalton. Amylase showed a pH optimum of 6.8, temperature 37 degree C, Km value of 2.0x10-3 g/mL, (lambda max) 540nm and incubation time for enzyme assay was 10 minutes. Increase in enzyme concentration showed a linear increase in amylase activity.
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