Lesson 6 of 8 / Biological molecules
Temperature and enzyme activity
Why does warming help only up to a point?
I can explain temperature effects and compare rates using a controlled enzyme investigation.
I can investigate and explain temperature and pH effects on enzyme reactions.Syllabus
Syllabus K327 / K328, Topic 3(e). Investigate and explain temperature and pH effects on enzyme reactions.
About 6 min
Make a guess. You are not marked.
Amylase is kept in a fridge at 4 degrees C, then warmed to 37 degrees C. What happens to its activity?
Show the answer
It rises again, because cold did not damage it.
Cold slows molecules so collisions are less frequent. The active site keeps its shape, so warming restores activity.
The key idea
Warming gives molecules more kinetic energy, so the frequency of effective collisions rises. Above the optimum, denaturation changes the 3D shape of the active site and the rate falls. Cooling usually slows activity without denaturing the enzyme.
Reaction rate is how quickly substrate is used or product is formed. For a fixed amount of reaction, a shorter completion time means a higher average rate.
Explore the idea
Shorter time, higher relative rate
30 degrees C: 60 s. Relative rate = 1/60 = 0.0167 per second. The same endpoint takes half as long as at 20 degrees C, so the relative rate is twice as large.
Original practice data, not published measurements. All mixtures use the same starting amounts, pH and endpoint. Dotted lines guide the eye; intermediate temperatures were not measured.
Explanation
Enzymes work only in solution. The enzyme and its substrate must be dissolved in water so that they can move and collide. An effective collision is a collision in which the substrate enters the active site and an enzyme-substrate complex forms.
At low temperature, enzyme and substrate molecules have little kinetic energy and move slowly. They collide less often, so fewer enzyme-substrate complexes form per unit time. As temperature rises, the molecules gain kinetic energy and move faster. The frequency of effective collisions increases, so the rate of reaction increases.
An enzyme has an optimum temperature at which its activity is greatest under the conditions tested. Beyond it, heat breaks the bonds that hold the enzyme in its 3D shape. The 3D shape of the active site changes, so the substrate is no longer complementary to it and cannot bind. The enzyme is denatured, fewer enzyme-substrate complexes form and the rate falls. The optimum is not the same for every enzyme.
Cold usually reduces activity without permanently changing the active site. Warming a cold enzyme appropriately can restore a higher rate. Heating a denatured enzyme and then cooling it does not usually restore its original function. Avoid saying that an enzyme is alive and can die.
To investigate temperature with amylase and starch, use water baths to bring enzyme and substrate separately to the chosen temperature before mixing. Keep pH, concentrations and volumes constant. At regular intervals, test a small drop of reaction mixture with iodine on a spotting tile. The endpoint is the first test where starch is no longer detected.
Use the same sampling interval and endpoint rule for every temperature, repeat each condition and compare means. If the same amount of starch is digested, 1/time is a relative measure of rate. An endpoint time is less precise when sampling intervals are long; shorter regular intervals improve the estimate.
Step by step
- 1
Control the comparison
Change temperature only. Keep pH and the amounts and concentrations of enzyme and substrate constant.
- 2
Measure the same endpoint
Use the same iodine sampling method to identify when starch is no longer detected.
- 3
Explain both sides of the pattern
Rising rate: more kinetic energy and a higher frequency of effective collisions. Falling rate at high temperature: the enzyme is denatured and the 3D shape of the active site changes.
Worked example
A shorter time means a faster reaction
Equal starch mixtures reach the same endpoint in 120 s at 20 degrees C and 60 s at 30 degrees C. How do the relative rates compare?
One way to explain it
The relative rates are 1/120 and 1/60 per second. The reaction at 30 degrees C is twice as fast by this measure because the same endpoint is reached in half the time. This comparison does not by itself identify the optimum temperature.
Why this answer works
- Use the same amount of reaction and endpoint.
- Compare reciprocals, rather than treating a larger time as a larger rate.
- Do not infer an optimum from only two temperatures.
Watch out for this
A student says: "An enzyme becomes denatured whenever it works slowly." What is wrong with this?
Show the answer
Low temperature can slow collisions while leaving the active site intact. A low rate alone does not establish denaturation; the cause and conditions matter.
Check your understanding
An enzyme is slow at 5 degrees C but becomes active when warmed to 30 degrees C. Which explanation fits?
Show the answer
Cold slowed collisions without permanently changing its active site.
The recovered activity supports a reversible effect of cooling rather than permanent denaturation.
Exam-style questions
Check your understanding
Invented data: amylase takes 120 s to digest starch at 20 degrees C, 40 s at 40 degrees C and 300 s at 60 degrees C. Which statement fits the data?
Show the answer
The rate is highest at 40 degrees C and falls at 60 degrees C.
The shortest time means the fastest rate, so 40 degrees C is fastest; 300 s at 60 degrees C is much slower.
Quick recall
Why does warming help only up to a point?