During protein purification, what term denotes enzyme activity per unit mass of total protein, used to quantify purification progress?

Difficulty: Easy

Correct Answer: Specific activity

Explanation:


Introduction / Context:
Protein purification workflows track both yield and purity. A key progress indicator is activity normalized to total protein, which allows comparison across fractions independent of volume or concentration differences.


Given Data / Assumptions:

  • Assaying a catalytic protein across purification steps.
  • Measuring total activity (units) and total protein (mg).


Concept / Approach:
Specific activity = activity units per milligram of total protein (U/mg). As purification proceeds, total protein usually decreases while the fraction that is the target enzyme rises, so specific activity should increase if the procedure is effective.


Step-by-Step Solution:

Measure activity in each fraction using a validated assay.Measure protein concentration and compute total mg.Compute specific activity = units / mg; compare step to step.


Verification / Alternative check:
Fold-purification can be calculated by dividing the specific activity at a given step by that of the crude extract; trends should align with SDS-PAGE band enrichment.


Why Other Options Are Wrong:

  • Relative activity: ambiguous without normalization to protein mass.
  • Purity ratio: non-standard term in enzymology.
  • All of these: only specific activity is the accepted definition.
  • Turnover frequency per cell: not a purification metric.


Common Pitfalls:
Confusing yield (total units) with purity (specific activity), or comparing activities without protein normalization.


Final Answer:
Specific activity.

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