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TOPIC 2.5 ENZYMES

U.1. Enzymes have an active site to which specific substrates bind.

Enzyme: An enzyme is built from proteins folded in complicated shapes and they are present throughout the body, the chemical reactions that develop inside of our bodies rely on the enzymes.

Substrate: A substrate is a molecule that acts with an enzyme. Enzymes catalyse chemical reactions involving the substrates. The substrate bonds with the enzyme active site, and that creates an enzyme-substrate complex.

Catalysis process.

1. In the first step the polar regions of amino acids attract the substrate and the active site of the enzyme.

2. The active site is the surface of an enzyme to which substrates bind and that catalyses the reaction.

3. Once the substrate has been bind, the reaction is catalysed.

4. The products are released and the enzyme is used again.

4. The products are released and the enzyme is used again. The lock and key theory

The lock and key theory

● This theory says that the substrate and the active site are specific to each other. They match each other in 2 ways.

are specific to each other. They match each other in 2 ways. Structurally: The structure of

Structurally: The structure of the active site is specific to the substrate. Substrates that don´t fit, don't react.

Chemically: Substrates that are not chemically attracted to the active site, won't react Induced-fit model

● If the lock and key theory was true then the enzyme could only catalyse one reaction, but in reality enzymes can catalyse more than one fractions. The induced fit theory says that as the substrate approaches the enzyme it provoqueis a conformational change in the active site, this meaning that it changes the shape in order for the substrate to fit.

● This stresses the substrate, reducing the activation energy of the reaction.

substrate, reducing the activation energy of the reaction. U.2. Enzyme catalysis involves molecular motion and the

U.2. Enzyme catalysis involves molecular motion and the collision of substrates with the active site.

1

The substrate binds to the active site. Some enzymes have two substrates that bind to different parts of the active site

substrates that bind to different parts of the active site 2 The substrates bind to the

2

The substrates bind to the active site change into different chemical substances, which are the products of

different chemical substances, which are the products of 3 The products separate from the active site,

3

The products separate from the active site, leaving it making it possible for more substrates to bind again.

● Enzyme activity is the catalysis of a reaction by an enzyme. These are the three steps

A substrate molecule can only bind to the active site if it moves very close to it. The getting together of a substrate molecule and an active site is known as collision. With most reactions the substrates are dissolved in water around the enzyme. Because of water being a liquid, the particles dissolved in it are in contact with each other and are in constant motion. Both substrates and enzymes are able to move, either way as substrates are typically known to be smaller than enzymes they tend to move faster.

Collisions between substrates and active sites occur because of random movement of the substrate and the enzyme. The successful collisions are those in which the substrate and active site are in site, allowing the binding to occur.

U.4. Enzymes can be denatured.

Enzymes, like other proteins can have their structure altered by certain conditions. This process called denaturation can be caused by high temperatures and either high or low pH.

When an enzyme is denatured, the active site is altered, so the substrate can no longer bind, and if it binds the reaction probably won't occur. In many cases denaturation causes enzymes

in

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insoluble a precipitate. were to and dissolved become form U.3. Temperature, pH and substrate concentration affect

U.3. Temperature, pH and substrate concentration affect the rate of activity of enzymes.

Enzyme activity is affected by temperature in two ways

1. In liquids, the particles are in constant random motion. When the liquid is heated the particles in it are given more kinetic energy. Both the enzymes and the substrates move faster in higher temperature thus increasing the the chance of a substrate molecule colliding with the active site of the enzyme. Enzyme activity increases.

2. When enzymes get heated, bonds of the enzyme vibrate, making them more weak, which increases the probability of the enzyme´s bonds to break. When the bonds inside the enzymes break, the structure of the enzyme changes, and with it, the active site. This change is permanent and is called denaturation. When the enzyme is denatured, it can no longer get back to its original state, which means it can’t catalyse reactions. As more enzymes get denatured enzyme activity falls, when the enzyme is completely denatured the activity stops altogether.

is completely denatured the activity stops altogether. Enzymes are affected by pH concentration. Most enzymes have

Enzymes are affected by pH

the activity stops altogether. Enzymes are affected by pH concentration. Most enzymes have an optimum pH

concentration.

Most enzymes have an optimum pH in which their activity is in its highest peak. If the pH is increased or decreased from that optimum, enzyme activity decreases and then stops altogether, when this happens the structure of the enzyme is altered, including its active site. Beyond a specific pH the structure is irreversible altered, this is another example of denaturation. Not all enzymes share the same optimum pH, actually the range is pretty large. This reflects the wide range of pH environment in which enzymes work. The image by the left side shows the wide range of pH optimus in different places where enzymes function

Enzyme

activity

is

affected

by

substrate

If the concentration of substrates increases, substrate-active site collisions will happen more frequently and the rate at which the enzyme catalyses the reaction will increase. However, after the binding of the substrate to an active site, the active site will be unavailable to other substrate molecules until a product is formed and released. As the concentration rises, more active sites will be unavailable. This means that the rate in which enzymes catalyse reactions get smaller as substrate concentration rises.

The graph below shows how the relationship between substrate concentration and enzyme activity works.

U.5

enzymes

industry.

are

and enzyme activity works. U.5 enzymes industry. are Immobilized widely used in In 1897 the Buchner

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widely

used

in

In 1897 the Buchner brothers, showed that an extract of yeast, containing no yeast cells, would convert sucrose into alcohol. More than 500 enzymes now have commercial uses, the graph below shows a classification of commercially useful enzymes.

below shows a classification of commercially useful enzymes. The enzymes used in industries are normally immobilized.

The enzymes used in industries are normally immobilized. This means the attachment of the enzymes to another materia or into aggregations, so that movement of the enzyme is restricted. There are many ways of doing this, attachment of enzymes in glass, trapping the enzymes in an alginate gel, or bonding them together.

Advantages of immobilized enzymes

Concentration of substrate can be increased as the enzyme is not dissolved.

Recycled enzymes can be used many times

O-

Separation of products is straightforward

Stability of the enzymes, as changes in the pH and temperature can be controlled

A.1. Methods of production of lactose free milk and its advantages.

Lactose is a sugar that is naturally present in milk, it can be converted into glucose and

galactose by the enzyme lactase: lactose → glucose + galactose.

Reasons for using lactase in the production of food.

1. Some people are lactose intolerant and cannot drink more than 250ml of milk per day, unless it's lactose reduced.

2. Galactose and glucose are sweeter than lactose, so less sugar needs to be added to sweet food containing milk, such as milkshakes and yogurts.

3. Bacteria ferments glucose and galactose more quickly than lactose, so the production of yogurt and cottage cheese is faster.

so the production of yogurt and cottage cheese is faster. The image in the left illustrates

The image

in

the

left illustrates the

process of producing lactose free milk

S.1. Design of experiments to test the effect of temperature, pH and substrate concentration on the activity of enzymes.

S.2. Experimental investigation of a factor affecting enzyme activity.

Catalyse is one of the most widespread enzymes, it catalyses the conversion of hydrogen peroxide, a toxic-by-product of metabolism, into water and oxygen.

1. You are going to investigate the independent variable, you need to decide:

● How you are going to vary it, example, with substrate concentration you would obtain a solution with the highest concentration and dilute it to get lower concentrations.

● What units will you use to measure the independent variable

● What range do you need for the independent variable, highest and lowest value.

2. The variable you use to find out how fast the enzyme catalyses is the dependent variable you need to decide:

● How are you going to measure it

● What unit should be used to measure the dependent variable.

● How many repetitions do you need to get reliable results.

3. You also need to be aware of the controlled variables., You need to decide

● What are the control variables

● How can you keep them constant

● What level should they be kept at, for example temperature.