(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,

Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
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Chapter1: Biochemistry: An Evolving Science
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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?
Transcribed Image 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?
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