PROBLEM 2: A biochemist dliscovers and purifies a new enzyme, generating a purification table below The step of protein purification Total Total Specific Total The multi- Total activity yicld (%)...


PROBLEM 2: A biochemist dliscovers and purifies a new enzyme, generating a purification table<br>below<br>The step of<br>protein<br>purification<br>Total<br>Total<br>Specific<br>Total<br>The multi-<br>Total<br>activity<br>yicld<br>(%)<br>cnzyme<br>fold<br>protcin<br>(mg)<br>maltifold<br>activity<br>increase in<br>purification<br>(units)<br>purity<br>Whole-cell lysate<br>8.4<br>LO<br>500<br>l00<br>4200<br>40% ammonium<br>22.2<br>95<br>2.64<br>9.64<br>180<br>3990<br>sulfate<br>Ion exchange<br>90.2<br>251<br>6.63<br>68<br>3787<br>55.7<br>Gel filtration<br>50<br>5000<br>to0<br>119<br>1.8<br>11.9<br>(a) From the information given in the table, calculate the specific activity of the enzyme,<br>total yield (%), multi-fold increase in purity, and total multifold purification after each<br>purification procedure.<br>b) Which of the purification procedures used for the enzyme is most effective (Le., gives<br>the greatest relative increase in purity)?<br>(c) Which step - ion-exchange or gel filtration chromatography - causes the greatest loss<br>of the protein.<br>(d) If you wanted to use only one purification step, which technique would<br>you<br>choose?<br>

Extracted text: PROBLEM 2: A biochemist dliscovers and purifies a new enzyme, generating a purification table below The step of protein purification Total Total Specific Total The multi- Total activity yicld (%) cnzyme fold protcin (mg) maltifold activity increase in purification (units) purity Whole-cell lysate 8.4 LO 500 l00 4200 40% ammonium 22.2 95 2.64 9.64 180 3990 sulfate Ion exchange 90.2 251 6.63 68 3787 55.7 Gel filtration 50 5000 to0 119 1.8 11.9 (a) From the information given in the table, calculate the specific activity of the enzyme, total yield (%), multi-fold increase in purity, and total multifold purification after each purification procedure. b) Which of the purification procedures used for the enzyme is most effective (Le., gives the greatest relative increase in purity)? (c) Which step - ion-exchange or gel filtration chromatography - causes the greatest loss of the protein. (d) If you wanted to use only one purification step, which technique would you choose?

Jun 11, 2022
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