A biochemist discovers and purifies a new enzyme, generating the purification table below. Procedure Total protein (mg)Activity (units) 1. Crude extract 20,000 4,000,000 2. Precipitation (salt) 5,000 3,000,000 3. Precipitation (pH) 4,000 1,000,000 4. lon-exchange chromatography 200 800,000 5. Affinity chromatography 50 750,000 6. Size-exclusion chromatography 45 675,000 (a) From the information given in the table, calculate the specific activity of the enzyme after cach purification procedure. (b) Which of the purification procedures used for this enzyme is most effective? (c) Which of the purification procedures is least effective? (d) Is there any indication based on the results shown in the table that the enzyme after step 6 is now pure? (e) What else could be done to estimate the purity of the enzyme preparation?

Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
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A biochemist discovers and purifies a new enzyme, generating the purification table below.
Procedure
Total protein (mg)Activity (units)
1. Crude extract
20,000
4,000,000
2. Precipitation (salt)
5,000
3,000,000
3. Precipitation (pH)
4,000
1,000,000
4. lon-exchange chromatography
200
800,000
5. Affinity chromatography
50
750,000
6. Size-exclusion chromatography
45
675,000
(a) From the information given in the table, caleulate the specific activity of the enzyme after
each purification procedure.
(b) Which of the purification procedures used for this enzyme is most effective?
(c) Which of the purification procedures is least effective?
(d) Is there any indication based on the results shown in the table that the enzyme after step 6
is now pure?
(e) What else could be done to estimate the purity of the enzyme preparation?
Transcribed Image Text:A biochemist discovers and purifies a new enzyme, generating the purification table below. Procedure Total protein (mg)Activity (units) 1. Crude extract 20,000 4,000,000 2. Precipitation (salt) 5,000 3,000,000 3. Precipitation (pH) 4,000 1,000,000 4. lon-exchange chromatography 200 800,000 5. Affinity chromatography 50 750,000 6. Size-exclusion chromatography 45 675,000 (a) From the information given in the table, caleulate the specific activity of the enzyme after each purification procedure. (b) Which of the purification procedures used for this enzyme is most effective? (c) Which of the purification procedures is least effective? (d) Is there any indication based on the results shown in the table that the enzyme after step 6 is now pure? (e) What else could be done to estimate the purity of the enzyme preparation?
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