Procedure step Volume (ml) Activity (units/ml. min) Total units Protein Concentr
ID: 168174 • 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
13
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
13
0.12
0.026
0.0035
Explanation / Answer
Purification of protein LDH (lactate dehydrogenase) involves 4 steps. The crude extract contained the protein concentration 31mg/mL. Dissolving in (NH4)2SO4 pellet is the second step and the protein concentration obtained was 1320mg/mL. The % of recovery of protein was 9%. Gel filtration was carried out in 3rd step and concentration of protein obtained was 21.38mg/mL and % of recovery was 0.036%. Ion - Exchange was carried out in 4th step and concentration of protein obtained was 13mg/mL and % of recovery was 0.026%. Comparitively purification was high during gel filtration among all steps and it was 0.0038. Therefore, form the data given in above table it can be concluded that LDH purificatio is most favored by Gel filtration step.
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