Table 4 gives the “Student Alex” CDNB assay data for the unknown protein. Again,
ID: 527900 • Letter: T
Question
Table 4 gives the “Student Alex” CDNB assay data for the unknown protein. Again, you can plot this data using Excel.
Table 4: “Student Alex” results for the crude lysate before purification of the GST-GFPS65T from the sample.
Reaction time (min)
A340 for 1/8 dilution of lysate
A340 for 1/4 dilution of lysate
A340 for ½ dilution of lysate
0
0
0
0
1
0.10
0.19
0.40
2
0.23
0.48
0.47
3
0.31
0.51
0.50
4
0.44
0.49
0.51
5
0.53
0.52
0.50
Using the “Student Alex” data shown in Table 4, plot A340 against time for each dilution using all of the data shown in Table 4.
(1 mark)
Now you will calculate the rate of reaction for each concentration of enzyme. For your calculations of rate you should not be using all of the data given in this table. You should only use the portions of the graphs that are linear and show Vmax for the enzyme. Clearly show your marking team which parts of the data you are using for your calculations. You can do this in one of two ways.
EITHER Re-plot the data, to show the points you are using to calculate the rate
OR Circle the region on your three-line plot that you are using to calculate the rate.
(2 marks)
Find the equation for the linear (Vmax) part of the curve/s you are using in Section 11b and show it on the plot.
(1 mark)
Reaction time (min)
A340 for 1/8 dilution of lysate
A340 for 1/4 dilution of lysate
A340 for ½ dilution of lysate
0
0
0
0
1
0.10
0.19
0.40
2
0.23
0.48
0.47
3
0.31
0.51
0.50
4
0.44
0.49
0.51
5
0.53
0.52
0.50
Explanation / Answer
1. For the 1/8th dilution of the lysate, the following plot shows the rate kinetics
Since the plot shows that almost all the points represent a linear plot, and hence for determining the rate of the reaction, the trendline was obtained considering all the points, the slope of the plot is: 0.1071 which is the rate of the reaction.
2. For the 1/4th dilution of the lysate, the following plot shows the rate kinetics
For the rate kinetics, considering only the linear points
From the plot the rate of the reaction = slope= 0.24 and Vmax=0.55
3. For 1/2 dilution
The linear portion plot is :
Cosidering the linear portion only the rate of reaction= 0.4 and Vmax=0.5
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