2 M O L E C U L A R B I O L O G Y

enzym xpimnts

Experimental investigation of a factor aecting enzyme activity.

There are many worthwhile enzyme experiments. constant if investigating the effect of

The method that follows can be used to substrateconcentration. [2]

investigate the effect of substrate concentration on

4 Predict whether the enzyme activity will

the activity of catalase.

change more if substrateconcentration is

Catalase is one of the most widespread enzymes. increased by 0.2 mol dm or if it isdecreased

It catalyses the conversion of hydrogen peroxide, by the same amount. [2]

a toxic by-product of metabolism, into water and

5 Explain why tissues such as liver must be

oxygen. The apparatus shown in gure 10 can be

macerated before investigating catalase

used to investigate the activity of catalase in yeast.

activity in them. [2]

The experiment could be repeated using the same

Safety goggles must be worn if this experiment

concentration of yeast, but different hydrogen

is performed. Care should be taken not to get

peroxide concentrations. Another possible

hydrogen peroxide on the skin.

investigation would be to assess the catalase

concentrations in other cell types, such as liver,

oxygen
kidney or germinating seeds. These tissues would

have to be macerated and then mixed with water

at the same concentration as the yeast.

measuring cylinder
yeast

three-way tap
1 Describe how the activity of the enzyme

water

catalase could be measured using the

apparatus shown in gure 10. [2]

2 Explain why a yeast suspension must always

be thoroughly stirredbefore a sample of it is water

0.8 mol dm

taken for use in an experiment. [2]

hydrogen peroxide

3 State two factors, apart from enzyme

concentration, that should bekept

▲ Figure 10 Apparatus for measuring catalase activity

▲ Figure 11 Enzyme experiment

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 2 . 5 e n z y M e s

d- q: Designing an experiment to nd the eect of temperature on lipase.

Lipase converts fats into fatty acids and glycerol. It 2 a) Explain how you would measure the

therefore causes a decrease in pH. This pH change dependent variable accurately. [2]

can be used to measure the activity of lipase.

b) State the units for measuring the

Figure 12 shows suitable apparatus.

dependent variable. [1]

tube contents mixed when both c) Explain the need for at least three

have reached target temperature

replicate results for eachtemperature

in this experiment. [2]

thermometer
3 a) List the control factors that must be

.v
/ ~

b)
kept

Explain
constant

how
in

these
this experiment.

control factors can

[3]

be kept constant. [2]

c) Suggest a suitable level for each

control factor. [3]

'-- ~
4 Suggest reasons for:

/ a) milk being used to provide a source of

thermostatically lipase milk mixed with

lipids in this experimentrather than

controlled sodium carbonate (an alkali)

vegetable oil. [1]

water bath and phenolphthalein

(a pH indicator)

b) the thermometer being placed in the

▲ Figure 12 Apparatus for investigating the activity of lipase

tube containing the larger, rather than

the smaller, volume of liquid [1]

Phenolphthalein is pink in alkaline conditions,

but becomes colourless when the pH drops to c) the substrate being added to the

7. The time taken for this colour change can be enzyme, rather than the enzyme to

used to measure the activity of lipase at different the substrate. [1]

temperatures. Alternatively, pH changes could

5 Sketch the shape of graph that you would

be followed using a pH probe and data-logging

expect from this experiment, with a

software.

temperature range from 0 °C to 80 °C on

1 a) State the independent variable in this the x-axis and time taken for the indicator

experiment and howyou would vary it. [2] to change colour on the y-axis. [2]

b) State the units for measuring the 6 Explain whether lipase from human pancreas

independent variable. [1] or from germinatingcastor oil seeds would

be expected to have the higher optimum

c) State an appropriate range for the

temperature. [2]

independent variable. [2]

Immbiizd nzyms

Immobilized enzymes are widely used in industry.

In 1897 the Buchner brothers, Hans and Eduard, showed that an

extract of yeast, containing no yeast cells, would convert sucrose into

alcohol. The door was opened to the use of enzymes to catalyse chemical

processes outside living cells.

Louis Pasteur had claimed that fermentation of sugars to alcohol could

only occur if living cells were present. This was part of the theory of

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