The effect of temperature on enzyme activity was determined
by pre-incubating the purified enzyme sample in 100 mM phosphate
buffer at different temperatures (25e60C) for 1 h, and residual
activity was measured using the standard assay conditions.
The effect of pH on enzyme activity was studied over a pH range of
4.0e10.0. The pH stability of the enzyme was determined by incubation
with different buffer systems at 50 C for 1 h, and residual
activity was measured. The following buffer systems (20 mM) were
used: sodium acetate buffer, pH 4.0e5.0; potassium phosphate
buffer, pH 6.0e7.0; TriseHCl buffer, pH 8.0; glycine-NaOH buffer,
pH 9.0e10.0.