Question

You are in the CHEM 464 lab and you are doing an expression and purification of a novel enzyme as part of an experiment. Shown below is the purification table: Procedure __________ Total Protein (mg) Activity (units) 1. Crude cellular extract 10,000 100,000 2. Ammonium sulfate precipitation 2,000 90,000 3. Affinity chromatography 17 51,000 4. Dialysis 13 39,000 (a) Calculate the specific activity of the enzyme of interest at each step. (b) Explain the principles behind each purification procedure in detail. How does it work? What characteristic of the enzyme is it using to purify it from the other biomolecules in the crude extract? (c) Did the dialysis appreciably improve the specific activity? What would be the reason for doing dialysis at the end of your enzyme purification?

          You are in the CHEM 464 lab and you are doing an expression and
purification of a
novel enzyme as part of an experiment. Shown below is the
purification table:
Procedure __________ Total Protein (mg) Activity (units)
1. Crude cellular extract 10,000 100,000
2. Ammonium sulfate precipitation 2,000 90,000
3. Affinity chromatography 17 51,000
4. Dialysis 13 39,000
(a) Calculate the specific activity of the enzyme of interest at
each step.
(b) Explain the principles behind each purification procedure in
detail. How does it
work? What characteristic of the enzyme is it using to purify it
from the other
biomolecules in the crude extract?
(c) Did the dialysis appreciably improve the specific activity?
What would be the
reason for doing dialysis at the end of your enzyme
purification?

Added by Kristin G.

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