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Please answer the following three questions involving this enzyme purification s

ID: 887121 • Letter: P

Question

Please answer the following three questions involving this enzyme purification scheme:

Fraction

Vol (mL)

Protein (mg)

Activity

Specific Activity

Crude lysate

200

2000

2646

1.32

A) differential centrifugation

200

1000

1323

1.32

B) Heat treatment

200

500

1200

2.4

C) Ion exchange

10

60

1100

18.3

D) Size exclusion

60

30

1000

33.3

E) Heparin Affinity

10

20

800

40

What step resulted in the largest loss of the enzyme of interest?

Fraction

Vol (mL)

Protein (mg)

Activity

Specific Activity

Crude lysate

200

2000

2646

1.32

A) differential centrifugation

200

1000

1323

1.32

B) Heat treatment

200

500

1200

2.4

C) Ion exchange

10

60

1100

18.3

D) Size exclusion

60

30

1000

33.3

E) Heparin Affinity

10

20

800

40

Explanation / Answer

I think you need to see the difference between specific activity of enzyme. If difference is more,it has resulted in largest lost of enzyme.

Ion exchange has largest difference from previous one which is 18.3-2.5 = 15.9

So answer : C) Ion exchange