*3.7. Thermo-Tolerance Properties of Aly08*

Purified Aly08 was placed at different temperatures for 1 h and then divided into two parts. One part was directly measured for its activity, while the other part was measured for its activity following incubation in an ice-bath for 30 min. In order to further observe whether the difference in temperature stability is related to boiling time, different boiling times (5, 10, 20, 30, 40, 50, and 60 min) were selected to evaluate the residual activity of the purified enzyme. After that the heat-treatment group was divided into two parts, one part was directly measured for its activity, while the other part was measured for its activity following incubation in an ice-bath for 30 min. To determine the effects of different temperatures on the activity recovery of heat-inactivated Aly08, the enzyme was immediately incubated for 30 min between 0 ◦C and 80 ◦C after boiling 5 min. Moreover, the enzyme was boiled for 5 min and further incubated at 0 ◦C for different times (0–60 min) to evaluate the effect of the time of low temperature treatment on the recovery of enzyme activity after it was heat treated.
