Procedure step Volume (ml) Activity (units/ml. min) Total units Protein Concentr
ID: 497044 • Letter: P
Question
Procedure step
Volume
(ml)
Activity (units/ml. min)
Total units
Protein Concentration
(mg/mL)
Specific Activity
(units/mg.min)
Percent recovery
(%)
Purification
Crude extract
36.0
1064
38304
31
34
Dissolved (NH4)2SO4 Pellet
6.0
579
3470
1320
0.0436
9.0
0.0013
Gel-Filtration
5.4
2.61
14.1
21.38
0.13
0.036
0.0038
Ion-Exchange
3.9
2.5
9.8
21.38
0.12
0.026
0.0035
How can I discuss the different methods of purification of LDH, based on the table of purification of LDH above?
Procedure step
Volume
(ml)
Activity (units/ml. min)
Total units
Protein Concentration
(mg/mL)
Specific Activity
(units/mg.min)
Percent recovery
(%)
Purification
Crude extract
36.0
1064
38304
31
34
Dissolved (NH4)2SO4 Pellet
6.0
579
3470
1320
0.0436
9.0
0.0013
Gel-Filtration
5.4
2.61
14.1
21.38
0.13
0.036
0.0038
Ion-Exchange
3.9
2.5
9.8
21.38
0.12
0.026
0.0035
Explanation / Answer
You can talk about the total activity in each step by looking into percent recovery
%recovery = [(activity of recovered fraction at each step x V) /(activity of starting material x V)]*100
At second step the activity of the recovered fraction is high when compared to others.
Also figures of purification fold indicates that total enzyme activity increases at every purification step
Concentration of enzyme can be high and also the interaction between enzyme and substrate seems to more
as there are more number of enzyme moles per unit volume.
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