Crude enzyme extract was purified by size exclusion chromatography
using column (10/50 mm) packed with Sephadex G-75.
The crude protein was first dialyzed by protein dialysis kit to
remove excess of salt. 2 mL of sample was loaded in the Sephadex
G-75 column and eluted by using 50 mM TriseHCL buffer at a flow
rate of 3.0 mL/5 min. A total of 22 fractions (3 mL each) were
collected. Fractions showing maximum enzyme activity were
pooled and then lyophilized; the concentrate was re-suspended in
the same buffer.