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Biology · 5 Enzymes · Paper 5/6 practical

Enzyme Activity. Find the optimum.

Investigate how temperature and pH change the rate of an enzyme reaction. Time how long amylase takes to digest starch (iodine end-point), or measure the oxygen catalase releases from hydrogen peroxide. Rate peaks at the optimum and falls to zero when the enzyme is denatured.

0610 Topic 5 — Enzymes temperature · pH · denaturation Paper 5/6 — Practical
Setup — set temperature/pH, then run and record the rate.
00.0 s
end-point

Variables

20
7
Keep the other variable constant — change only the one you're investigating.

Live readouts

Time to digest
Rate
Relative activity
Optimum
Above the optimum the enzyme's active site changes shape (denatures) and the rate falls to zero.

Results table

Record the rate at several values.

Rate vs temperature

📋 Method (Cambridge practical procedure)
  1. Amylase: mix amylase and starch solutions in a tube held in a water bath at the chosen temperature (or with a pH buffer).
  2. Every 30 s remove a drop and add it to iodine on a spotting tile.
  3. Record the time when the iodine stops turning blue-black — all the starch has been digested. Rate = 1 ÷ time.
  4. Catalase: add hydrogen peroxide to potato/liver and measure the oxygen given off (gas syringe or counting bubbles per minute).
  5. Repeat at a range of temperatures (or pH values) and plot rate against the variable.
⚠ Control variables & precautions
  • Keep volumes and concentrations of enzyme and substrate the same each time.
  • Allow the tubes to reach the water-bath temperature before mixing.
  • Use a buffer to fix the pH when investigating temperature, and a fixed temperature when investigating pH.
  • Repeat each value and take a mean; rate = 1/time so faster reactions give a higher rate.
🎯 Syllabus reference (0610)
  • 5 Enzymes — investigate and describe the effect of temperature and pH on enzyme activity; explain in terms of kinetic energy, collisions, active site and denaturation.

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