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A biochemist discovers and purifies a new enzyme, generating the purification table below. Procedure 1. Crude extract 2. Precipitation (salt) 3. Precipitation (pH) 4. Ion-exchange chromatography 5. Affinity chromatography 6. Size-exclusion chromatography Total protein (mg) 20,000 5,000 4,000 200 50 45 Activity (units) 4,000,000 3,000,000 1,000,000 800,000 750,000 675,000 a) From the information given in the table, calculate the specific activity of the enzyme after each purification procedure. b) Which of the purification procedures used for this enzyme is most effective (i.e., gives the greatest relative increase in purity)? 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? What else could be done to estimate the purity of the enzyme preparation?

A biochemist discovers and purifies a new enzyme, generating the purification table below. Procedure 1. Crude extract 2. Precipitation (salt) 3. Precipitation (pH) 4. Ion-exchange chromatography 5. Affinity chromatography 6. Size-exclusion chromatography Total protein (mg) 20,000 5,000 4,000 200 50 45 Activity (units) 4,000,000 3,000,000 1,000,000 800,000 750,000 675,000 a) From the information given in the table, calculate the specific activity of the enzyme after each purification procedure. b) Which of the purification procedures used for this enzyme is most effective (i.e., gives the greatest relative increase in purity)? 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? What else could be done to estimate the purity of the enzyme preparation?

Biochemistry
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
Section: Chapter Questions
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1. A biochemist discovers and purifies a new enzyme, generating the purification table below. Procedure 1. Crude extract 2. Precipitation (salt) 3. Precipitation (pH) 4. Ion-exchange chromatography 5. Affinity chromatography 6. Size-exclusion chromatography Total protein (mg) 20,000 5,000 4,000 200 50 45 Activity (units) 4,000,000 3,000,000 1,000,000 800,000 750,000 675,000 a) From the information given in the table, calculate the specific activity of the enzyme after each purification procedure. b) Which of the purification procedures used for this enzyme is most effective (i.e., gives the greatest relative increase in purity)? 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? What else could be done to estimate the purity of the enzyme preparation?
Transcribed Image Text:1. A biochemist discovers and purifies a new enzyme, generating the purification table below. Procedure 1. Crude extract 2. Precipitation (salt) 3. Precipitation (pH) 4. Ion-exchange chromatography 5. Affinity chromatography 6. Size-exclusion chromatography Total protein (mg) 20,000 5,000 4,000 200 50 45 Activity (units) 4,000,000 3,000,000 1,000,000 800,000 750,000 675,000 a) From the information given in the table, calculate the specific activity of the enzyme after each purification procedure. b) Which of the purification procedures used for this enzyme is most effective (i.e., gives the greatest relative increase in purity)? 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? What else could be done to estimate the purity of the enzyme preparation?
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