Enzymes are crucial to proper metabolic function, and ultimately life.

That is why you

need to know the conditions which affect enzyme activity. There are several properties
which can alter enzyme function:

1. Temperature
2. Inhibitors (competitive and non-competitive)
3. pH
4. Substrate concentration.

Enzymes are proteins, so have a delicate tertiary structure that enables that
enzyme's adequate function. High temperature or pH would alter its tertiary structure.
Inhibitors would bind to its active site, preventing substrates from doing so. This results
in no enzyme-substrate complexes being formed. Let's have a closer look at these
properties individually.

Temperature

Increasing temperature results in a higher rate of activity, up to a certain point where the
enzyme becomes denatured. A high temperature causes the molecule
to vibrate, breaking the weak bonds that hold it together, and changing the
structure of the enzyme. This process is denaturation. The point at which this
happens is usually around 50 - 60 degrees Celsius.

Denatured enzymes don't work. Look at this graph (click to enlarge) to understand the
relationship between enzyme activity and temperature:

Inhibitors

Inhibitors are molecules which interfere with the substrate binding to the active site of an
enzyme, slowing down or stopping the reaction. These may be reversible or non-
reversible inhibitors. The reversible inhibitors can be competitive or non-competitive.
Competitive inhibitors have a similar 3D shape to the substrate, hence they can bind
to the active site of the enzyme, preventing the substrate from doing so. It's easy to
picture:

The competitive inhibitors compete (as you'd expect) with the substrate for the active
sites of the enzymes. If more substrate is added, then the inhibitors' effect will be
diminished. This is what the graph looks like (make sure you can recall this):

Non-competitive inhibitors on the other hand bind to the enzyme at a site away from
the active site. All good? No, because that results in the enzyme's shape changing.
This means the substrate can no longer bind to the active site. Unlike the case of
competitive inhibitors, changing the substrate concentration will not have an effect on
the rate of reaction. Here is a comparison diagram (learn this):
Don't worry about the V (velocity), check your textbook for other graphs. If you're a
video sort of person, here is a nice one:

pH

Binding of the substrate to the enzyme depends on a close match

between shape and charge. The pH is a measure of the concentration of H+ ions
versus OH- ions. As you can see, these are positively or negatively charged, so a really
high or really low pH can disrupt enzyme function. All enzymes have a specific optimal
pH at which they work best. This differs between enzymes. For example, while most
enzymes work best at a pH of 7.35 (that is halfway between 1 and 14 - 1 is most acidic,
14 is most basic), pepsin in the stomach acid works best at a pH of 3.

Substrate concentration

This topic is a matter of common sense. However, you must use A level language. Here
it goes.

Common sense version: More substrate results in more reactions, so rate of reaction
goes up. Of course, when all enzymes are working all the time, adding even more
substrate will not increase the rate of reaction, unless more enzymes are added.
Catalase in potatoes
What is the optimum temperature for catalase from a potato? What temperature does it become denatured?
Peter Nicholls and John Hewitson, with assistance from Professor Peter Nicholls of Essex University
What we expect in an enzyme reaction is that it goes faster and faster as temperature is increased until a
temperature is reached at which the enzyme is denatured (its shape is changed and therefore its activity is
destroyed) before much reaction can be measured.
A second question is "How much does catalase respond to an increase in temperature up to the point at which it is
denatured?". For most enzyme catalysed reactions, there is approximately a doubling of the rate of reaction for every
10C rise. This is called a Q(10) of 2 and most enzymes have a Q(10) between 1.5 and 2.5. The strange thing about
catalase is that it has a Q(10) of less than 1.2 (between 0 and 40C). This means that temperature has hardly any
effect on the rate of reaction with catalase. In other words, there is hardly any increase in the rate of reaction as the
temperature increases. It turns out that the rate at which catalase can work is limited by the rate at which the
substrate (peroxide) can diffuse into the "active site" of the enzyme and this diffusion is a physical rather than a
chemical process (and is therefore much less affected by temperature).

What gas was produced by the breakdown of hydrogen peroxide?

Oxygen gas was produced

Describe the test that was performed in order to identify the gas.
A glowing splint of a match was lit, blown out then inserted into the test tube. The match relighting in the test
tube indicates oxygen gas is present.

Can hydrogen peroxide be broken down by catalyst other than those found in a living system?
Hydrogen peroxide can be broken down by manganese dioxide because it has catalytic properties. It is
unstable which makes it very reactive. It even breaks down in the presence of light. It increases the rate of
reaction without being changed. Sand however is not able to break it down because it contains no catalytic
properties.

Explain how temperature affected the enzymes function

Increasing the temperature increased the rate of reaction. There is a higher energy when heated. The enzyme
was able to catalyze the reaction more quickly. This is only until the point until denaturation. At 40 degrees, the
enzyme would experience denaturation causing the rate of reaction to drop. The enzyme would be damaged
and not be able to perform the same way.
How did particle size affect the rate of reaction?
Smaller size of particles increased the rate of reaction because smaller particles consume less energy than
larger ones to break down molecules, therefore the reaction would happen faster. Larger particles decreased
the rate of reaction because they require more energy to break down.

Explain why there is a difference in the rates of reaction between the liver and the potato
Liver contains more of the enzyme catalase, which breaks down hydrogen peroxide. Liver contains more
because it detoxifies substances in the body. A larger amount of catalase lowers the activation energy,
therefore speeds up the rate of reaction. The potato contains less of the enzyme catalase, therefore requires
more activation energy, slowing down the rate of reaction.

Show the fully labeled balanced chemical equation for the decomposition of hydrogen peroxide
2 H2O (aq) (catalase)> 2 H2O (l) + O2 (g)
hydrogen peroxide enzyme water oxygen gas

CONCLUSION AND FINAL INFORMATION

Conclusion:

The enzyme catalase is a common enzyme in animal and plant tissues. Catalase can separate two
hydrogen peroxide molecules into two water molecules and an oxygen molecule. This is
anIMPORTANT biochemical reaction because hydrogen peroxide can be dangerous because it
creates oxygen radicals that are desperate for electrons. We believed we could observe what type
of potato (boiled or frozen) would have a higher amount of catalase by measuring the bubbles
(O2) produced at the top of the potato in hydrogen peroxide solution.

In this lab, we measured the rate of enzymatic activity of catalase in boiled potatoes versus
frozen potatoes. We did this by putting 1 cm cubes of boiled, frozen, and raw potatoes in
graduated cylinders with 10 mL of hydrogen peroxide. After 2.5 minutes, we recorded how
much oxygen bubbles were produced from the enzymatic reaction of catalase of each potato
type: frozen, boiled, and raw (control group). We performed five trials of this.

We obtained the following results for the amount of oxygen bubbles produced: raw(1)-3.0 mL,
raw(2)-5.0 mL, raw(3)-3.0 mL, raw(4)-3.2 mL, raw(5)-2.9 mL; frozen(1)-6.0 mL, frozen(2)-5.0
mL, frozen(3)-4.0 mL, frozen(4)-4.1mL, frozen(5)-3.5 mL; boiled(1)-2.0 mL, boiled(2)-1.0mL,
boiled(3)-.5 mL, boiled(4)-1.6 mL, boiled(5)-1.1 mL. The average mL of O2 produced for raw
potatoes was 3.42 mL with a possible error of +/- .89 mL. The average mL of O2 produced for
frozen potatoes was 4.52 mL with a possible error of +/- .99 mL. The average mL of O2 for
boiled potatoes was 1.24 mL with a possible error of +/- .58 mL.

From looking at the results, we concluded that frozen potatoes have a higher rate of enzymatic
activity of catalase when compared to boiled potatoes. We concluded this because more O2
bubbles (about 3 mL) were produced on average for the frozen potatoes than the boiled
potatoes. Something we do not understand from our lab is why the frozen potato had more
enzymatic activity compared to the raw potato control group. On average the frozen potatoes
produced one more milliliter of O2 bubbles. It would seem that the raw potato would have more
catalase because it was not tampered with compared to the frozen potato. The freezing of the
potato might have enhanced or created more catalase in the potato. There is almost a possible
error one mL of oxygen bubbles for both raw potatoes and frozen potatoes, so raw potatoes could
have just as much catalase as frozen potatoes.

Our lab included many possible errors including the following:

1. When we dropped the potato into the solution, it caused displacement of the hydrogen
peroxide because of the space the potato took up. We did not pay attention to this
scenario the first three trials, but the last two trials we took note of the displacement.
Also, not all the potatoes were exactly 1 cm cubes, so they did not all create a
displacement of 1 mL (because 1 cm^3=1 mL). This creates inaccuracy in our results
because mL of bubbles we recorded may have been displacement of hydrogen peroxide.
2. The scale we used was faulty and the mass of the potatoes continued to change after
leaving the potato in there for at least five seconds. This may have caused some of our
masses of the potatoes to be inaccurate, and therefore some of the masses may have been
further away from the average of the masses of potatoes which could cause much more or
much less O2 bubbles to be produced.
3. We used the same three graduated cylinders for each trial, and did not have a great
amount of time to perform the lab, so the graduated cylinders may have not been dried
completely, which could cause error.
4. It was hard to pour exactly 10 mL of hydrogen peroxide into the 100 mL graduated
cylinder. More or less than 10 mL of hydrogen peroxide could alter results because
maybe there would be more or less hydrogen peroxide for the catalase to react with.

Possible improvements for the lab include:

1. A better way of measuring oxygen produced (although we do not know of any better
options available). Looking at the amount of oxygen produced by just looking at bubbles
with the bare eye is not very scientific. Some oxygen could have escaped into the air
because it is a gas.