(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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Step 1: Identify the least effective purification procedure from the table. Show more…
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A biochemist discovers and purifies a new enzyme, generating the purification table below. Purification Step | 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. Ion-exchange chromatography | 200 | 800,000 5. Affinity chromatography | 50 | 750,000 6. Size-exclusion chromatography | 45 | 675,000 a. From the information in the table, calculate the specific activity of the enzyme after each purification step. b. Which of the purification steps used for the enzyme is the most effective (i.e., gives the greatest relative increase in purity)? c. Which of the steps 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 effectiveness of the purification procedure, the purity of the final sample, and estimate the molecular weight of the enzyme?
Sri K.
Evaluating the purification of an enzyme.Âą An enzyme believed to be involved in type II diabetes is being purified to serve as a target in a drug development research project. At each step of the purification process, the total amount of protein present is quantified, and the activity of the enzyme is measured: Step # Technique 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 Ion exchange chromatography 200 800,000 5 Affinity chromatography 50 750,000 6 Size-exclusion chromatography 45 675,000 a. What is the specific activity after each purification step? b. Which purification step seems to have been the most effective (i.e., provided the greatest relative increase in purity)? Explain your reasoning. c. Which of the steps seems to have been the least effective? Explain briefly. d. Based on the data provided here, what can you say about the purity of the enzyme after the completion of step 6? What else could be done estimate the purity of the enzyme?
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