why TA of alcohol precipitation is used as TA of purified protein.

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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I am not understanding why TA of alcohol precipitation is used as TA of purified protein. I provided 2 photos, 1 with the questions and the correct answer, and the other photo is a screenshot of the explanation. 

mg
Therefore, the purification step with the greatest total activity would also have the highest
yield. The total activity of the target enzyme was highest with the alcohol precipitation
method:
u
:200 u
mg
TA of alcohol precipitation = 200 mg × 1.0-
The ratio of purified protein activity to unpurified protein activity is:
TA of purified protein
Yield %
х 100
TA of unpurified extract
200u
Yield %
х 100 — 0.5 х 100 — 50%
400u
Therefore, a maximum of 50% of Protein X was purified from the cell-free extract.
(Choice A) This value is the specific activity achieved with the unpurified extract.
(Choices B and D) These values represent the ratios of total purified protein to unpurified
protein obtained with alcohol precipitation and column chromatography, respectively. Yield
is determined by calculating ratios of total activity instead of total protein.
Educational objective:
Specific activity (u/mg), which reflects protein purity, is the ratio of the activity of the target
protein and the total protein. The total activity is the product of the total protein and the
specific activity. Yield is determined by dividing the total activity of the purified protein by the
total activity of the unpurified extract.
Transcribed Image Text:mg Therefore, the purification step with the greatest total activity would also have the highest yield. The total activity of the target enzyme was highest with the alcohol precipitation method: u :200 u mg TA of alcohol precipitation = 200 mg × 1.0- The ratio of purified protein activity to unpurified protein activity is: TA of purified protein Yield % х 100 TA of unpurified extract 200u Yield % х 100 — 0.5 х 100 — 50% 400u Therefore, a maximum of 50% of Protein X was purified from the cell-free extract. (Choice A) This value is the specific activity achieved with the unpurified extract. (Choices B and D) These values represent the ratios of total purified protein to unpurified protein obtained with alcohol precipitation and column chromatography, respectively. Yield is determined by calculating ratios of total activity instead of total protein. Educational objective: Specific activity (u/mg), which reflects protein purity, is the ratio of the activity of the target protein and the total protein. The total activity is the product of the total protein and the specific activity. Yield is determined by dividing the total activity of the purified protein by the total activity of the unpurified extract.
Question 8
Activity assays were performed to determine the best purification method for a cell-free
extract of Protein X. The results of the assays are shown below.
1.0 T
Alcohol precipitation
Native PAGE
-- Extract
Column chromatography
---.
0.8 +
0.6+
0.4+
0.2 +
200
400
600
800
Total protein (mg)
Based on the data, what was the highest purification yield of Protein X?
A. 0.5% (10%)
B. 25% (21%)
C. 50% (50%)
D. 75% (17%)
Specific activity (u/mg)
Transcribed Image Text:Question 8 Activity assays were performed to determine the best purification method for a cell-free extract of Protein X. The results of the assays are shown below. 1.0 T Alcohol precipitation Native PAGE -- Extract Column chromatography ---. 0.8 + 0.6+ 0.4+ 0.2 + 200 400 600 800 Total protein (mg) Based on the data, what was the highest purification yield of Protein X? A. 0.5% (10%) B. 25% (21%) C. 50% (50%) D. 75% (17%) Specific activity (u/mg)
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