Enzymes

Enzy mes have an active site to which specific substrates bind.

 Active site: The area or the pocket on the enzyme where the substrate binds.
 Enzy me: Proteins that catalyze chemical reactions (increase the rate by lowering the activation
energy)
 Each enzyme catalyzes a specific reaction for a specific substrate
 Enzy mes are not used up during the chemical reactions
 Enzy mes are very specific because both the enzyme and the substrate possess specific complementary
shapes that fit into one another.
 The binding of the substrate to the enzyme causes the chemical bonds of the substrate to weaken.
 This eventually causes the reactions that take place that form the products.
 After the products are released, the enzyme can bind to another substrate, because enzymes are not
used up in these chemical reactions.

Enzyme catalysis invol ves molecular motion and the collision of substrates with the acti ve site.

 When a substrate comes close to the active site of the enzyme, it can collide and bind to the active site
of the enzyme
 Since the substrate is dissolved in water around the enzyme, the substrates and enzymes are in
continuous motion
 The direction and movement is constantly changing and is random
 Collisions occur at random between the substrate and enzyme
 Successful reactions only occur if the substrate and the active site of the enzyme are correctly aligned
and the collide with sufficient KE

∑ - Temperature, pH and substrate concentration affect the rate of acti vity of enzymes.

Temperature

 When heat is added to a liquid the particles speed up, thus giving them more kinetic energy.
 In a liquid that contains substrates and enzymes, the increase in kinetic energy will cause more
collisions between substrates and enzymes thereby increasing enzyme activity and reaction rates.
 However, as temperature increases and becomes too high, the bonds of the enzyme begin to vibrate
and eventually break.
 This causes the enzyme to lose its 3D shape, including the shape of the active site.

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  When the enzyme loses its shape and can no longer catalyze reactions, the enzyme is said to be
denatured.
 When the enzymes in solution become denatured, the reaction rate decreases dramatically.
 Enzy me denaturation is usually permanent.

The optimu m rate of reaction is when the graph reaches the top of the curve which is around 40ºC for most
enzymes.

pH:

 pH is dependent on the number of H+ ions compared to the number of OH- ions.

 When a solution has a high number of H+ ions the solution it is said to have a low pH (acid). If a
solution has a high number of OH- ions the solution is said to have a high pH (base).
 Enzy mes have an optimu m pH at wh ich they work the best.
 When deviations occur away from this optimu m pH, the enzyme’s activity or reaction rate decreases.
 When the pH moves too far away from the enzy me’s optimu m pH, the enzyme will lose its shape and
denature, drastically decreasing enzyme activity.
 For example, the optimum pH for the enzy me pepsin is around 2-3. If the pH increases to 5 or 6, the
enzyme loses its ability to catalyze reactions (the breakdown of proteins in the stomach).
 Most enzymes have an optimu m pH close to neutral (7) pH

Substrate Concentration

 With a fixed amount of enzymes, as substrate concentration increases, the rate of reaction will
increase, because more collisions between enzymes and substrates will occur.
 However, as substrate-level increases, more and more enzy me active sites are being filled.
 At a certain substrate concentration, all active sites on the enzymes are being used or are filled.
 At this point, the reaction rate levels off and remains constant.
 Adding more substrate to the reaction will not increase the reaction rate. The reaction rate can only be
increased with the addition of more enzy mes.

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∑ - Enzymes can be denatured.

 Denaturation is a structural change in a protein (usually enzymes) that results in the loss (usually
permanent) of its biological properties. When an enzyme denatures, the bonds that hold together its
three-dimensional shape begin to vibrate and eventually break. This causes the enzyme to unfold and
lose its shape, thereby eliminating the enzyme’s ability to catalyze reactions.

Design experiments to test the effect of temperature, pH and substrate concentration on the acti vi ty of
enzymes.

https:// www.youtube.com/ watch? v=bG-XCG46t80

Important terms:
Accuracy: means how correct your results are (quantitative). For example counting bubbles to
measure the rate of enzyme activity when testing the effect of catalase on hydrogen peroxide is
not an accurate method.
H2O2 H2O + O2
Why? Because the sizes of bubbles are not equal and bubbles may contain other gases not only
Oxygen. To improve accuracy you can use a more sensitive measuring equipment (like a gas
syringe or a measuring cylinder in this example)
Reliability: means how similar the results are after several repeats.
To improve reliability you should increase repeats and get the average.
Validity: means how correct and fair is your process, to increase validity keep all other factors
constant.
Controlled variables: are the factors kept constant in an experiment (validity)

 Independent variable: Variable changed by the experimenter (placed on the x-axis) ex.
Time, days, years, light intensity, PH, temperature.
• Dependent variable: Variable measured by the experimenter (placed on the y-axis) is
the factor you are testing and measuring.
ex. Mass, volume, # of individuals, conc. of gases, pulse rate.

School sheet page 27

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∑ - Immobilized enzymes are wi dely used in industry.

 More than 500 enzy mes are now used for commercial purposes
 The majority of these enzymes used in industry are immobilized; meaning they are attached to another
material (such as glass) or grouped together (in a calcium alginate gel)
 The benefits of using immobilized enzy mes are as follows

 Convenience – only small amounts of proteins dissolve in the reactions leaving only solvent and the
products. This means the enzymes and products can be easily separated
 Economics – The immobilized enzy mes can be easily removed and reused, saving money.
Eg. Particular useful in the removal of lactase in the production of Lactose-Free Milk.
 Stability – Immobilized enzy mes generally have a greater thermal and chemical stability than the
soluble form of the enzy me
https://www.youtube.com/watch?v=36RXIjHMC6g

Methods of production of lactose-free milk and i ts advantages.

 Lactose is a disaccharide sugar present in milk composed of monosaccharides glucose and galactose.
 Lactase is the enzyme that breaks down lactose into its two monosaccharides.
 Humans are born with the ability to digest milk (lactase produced) but as we grow older, most humans
lose the ability to produce lactase in significant amounts.
 If the lactose is broken down in milk before it is consumed, people that are lactose intolerant can drink
the milk.
 Some types of yeasts produce lactase.
 Biotechnology companies can culture these yeasts and remove the lactase.
 Milk is treated with lactase before distribution, allowing lactose-intolerant people to consume milk
and milk products.

School sheet page 26

https://www.youtube.com/watch?v=P7e9Mj9ATpQ