Enzyme Reaction Lab Report
Figure 28: Lineweaver-Burk plot for evaluation of kinetic constants (Km and Vmax) of extracellular purified pectinase.
3.8.3. Effect of temperature of the reaction:
The effect of the temperature of the reaction on the activity of the purified enzyme was carried out by make the enzymatic reaction for 10 minutes at different temperature 25,30,35,40,45,50,60 and 70°C using an enzyme protein 0.1mg/reaction mixture and substrate concentration of 15 mg/reaction mixture, using a control of previously heated enzyme solution in the reaction.
The data recorded in (table 27) and (figure 29) illustrate the effect of temperature of the reaction on the pectinase activity, it showed that the pectinase activity increased by increasing the reaction temperature reaching its maximum value at 45°C, it reach 49.39U/mg protein. After that any rise of temperature makes loss in activity gradually. Therefore, in the coming experiments the temperature of the reaction had to be adjusted at 45°C. …show more content…
Effect of pH of the reaction:
The impact of the pH of the reaction on the activity of the pure pectinase was studied. The pH range of 3.5to 8 was used (acetate buffer, 0.1M; phosphate buffer, 0.1M).all the other reaction conditions were at their optimum values as determined from the previous experiments.
Table 27: effect of the reaction temperature on the activity of the purified pectinase obtained from Brevibacillus borstelensis MSB6.
Temperature(°C) Pectinase activity
(U/mg protein) Relative activity (%)
25 34.17 69.19
30 40.78
3.8.3. Effect of temperature of the reaction:
The effect of the temperature of the reaction on the activity of the purified enzyme was carried out by make the enzymatic reaction for 10 minutes at different temperature 25,30,35,40,45,50,60 and 70°C using an enzyme protein 0.1mg/reaction mixture and substrate concentration of 15 mg/reaction mixture, using a control of previously heated enzyme solution in the reaction.
The data recorded in (table 27) and (figure 29) illustrate the effect of temperature of the reaction on the pectinase activity, it showed that the pectinase activity increased by increasing the reaction temperature reaching its maximum value at 45°C, it reach 49.39U/mg protein. After that any rise of temperature makes loss in activity gradually. Therefore, in the coming experiments the temperature of the reaction had to be adjusted at 45°C. …show more content…
Effect of pH of the reaction:
The impact of the pH of the reaction on the activity of the pure pectinase was studied. The pH range of 3.5to 8 was used (acetate buffer, 0.1M; phosphate buffer, 0.1M).all the other reaction conditions were at their optimum values as determined from the previous experiments.
Table 27: effect of the reaction temperature on the activity of the purified pectinase obtained from Brevibacillus borstelensis MSB6.
Temperature(°C) Pectinase activity
(U/mg protein) Relative activity (%)
25 34.17 69.19
30 40.78