3.6 ENZYMES + 7.

6 ENZYMES (HL)

3.6 ENZYMES

ASSESSMENT STATEMENTS
3.6.1: Define enzyme and active site. 3.6.2: Explain enzymesubstrate specificity. 3.6.3: Explain the effects of temperature, pH and substrate concentration on enzyme activity. 3.6.4:Define denaturation. 3.6.5:Explain the use of lactase in the production of lactose-free milk.

WHAT ARE ENZYMES?

They are globular proteins. They are biological catalysts. They speed up metabolic reactions but remain unchanged. They are specific. Enzymes have an active site that fits the substrate precisely.

Substrate

Substrate

Enzyme
Product

OR
Enzyme Product Product

Substrate

OR
Substrate
Enzyme

Product

ENZYME-SUBSTRATE SPECIFICITY
The active site is the specific region of an enzyme to which the substrate or substrates bind.

Each enzyme will only act on substrates that will fit into its active site. Each enzyme acts on only one (or a limited number) of substrates.

LOCK AND KEY MODEL

Enzyme

Substrate: shape is complementary to shape of active site Active site

DENATURATION

Denaturation is a structural change in a protein that results in the loss (usually permanent) of its biological properties. Changes in the chemical environment of the enzyme can lead to a shape / conformational change in the protein. This could lead to a change in the shape of the active site. This may interfere with the binding of the substrate with the active site.

WHAT CAN CAUSE DENATURATION?

Treat with high temperature or strong acid/alkali
Active site no longer fits round substrate

TEMPERATURE
Every enzyme works within a range of temperatures. Enzymes work best at their optimum temperature. Increase in temperature increases chance of enzyme- substrate collisions so enzyme activity increases. Further increase in temperature can increase molecular motion leading to disruption of intermolecular interactions.

Rate of reaction increases as temperature increases and peaks at the optimum temperature. Increasing the temperature further decreases the rate of reaction until all enzymes are denatured and rate of reaction reaches zero.

PH
Every enzyme works within a certain range. Each enzyme has an optimum pH. Altering pH can alter intermolecular interactions within the enzyme or within the active site. Rate of reaction decreases as the pH moves further away from the optimum pH.

SUBSTRATE CONCENTRATION
As substrate concentration increases, the rate of reaction increases. The increase in rate of reaction is proportional to the increase in substrate concentration until a point where all active sites are occupied. The rate of reaction will not increase any further beyond this point but rather remain constant.

LACTOSE INTOLERANCE

Lactose intolerance results from the inability to break down lactose, the sugar found in milk

Lactose intolerance is high in some human populations: Asian / African / native American and Australian aboriginal populations.

LACTOSE-FREE MILK

Lactase is used to produce lactose-free / low-lactose milk. Source of lactase is usually yeast but there are many sources such as bacteria and moulds.

HOW IS LACTOSE-FREE MILK MADE?

Milk is passed over immobilized lactase (lactase bound to inert substance). This results in an increase in the sweetness of milk. There is no need to add extra sugar in the manufacture of flavoured milk drinks / frozen desserts.

7.6 ENZYMES (HL)

ASSESSMENT STATEMENTS

7.6.1: State that metabolic pathways consist of chains and cycles of enzyme-catalysed reactions. 7.6.2: Describe the induced-fit model. 7.6.3: Explain that enzymes lower the activation energy of the chemical reactions that they catalyse. 7.6.4: Explain the difference between competitive and noncompetitive inhibition, with reference to one example of each. 7.6.5: Explain the control of metabolic pathways by endproduct inhibition, including the role of allosteric sites.

METABOLIC PATHWAYS

Metabolic pathways consist of chains and cycles of enzyme-catalysed reactions.

EXAMPLE 1
Examples of a metabolic pathway, that is a chain, are glycolysis in cell respiration and photophosphorylation in photosynthesis.

8.1,8.2

EXAMPLE 2

8.1,8.2

Example of a metabolic pathway, that is a cycle, are Krebs cycle in cell respiration and Calvin cycle in photosynthesis.

ENZYME SUBSTRATE SPECIFICITY

The active site of an enzyme binds to specific substrate. The shape of the active site and substrate fit / complement each other (lock and key model). The chemical properties of substrate and enzyme attract due to opposite charges.

INDUCED FIT MODEL

The enzyme / active site is not rigid and substrate can induce slight changes in shape. This allows substrates of similar structure to bind with same enzyme. Induced fit causes weakening of bonds in substrate to lower activation energy.

ACTIVATION ENERGY
Activation energy is the energy that is required for a chemical reaction to start. This is often in the form of heat energy, which increases the movements of the substrate molecules or ions.

ACTIVATION ENERGY
Energy Key: original reaction with enzyme

Progress of reaction

STEP BY STEP
substrate binds / approaches active site shape of active site changes bonds in substrate weaken activation energy decreases explains broad specificity of some enzymes eg proteases

ACTIVATION ENERGY LOWERED

enzyme

binds to substrate lowers activation energy by weakening bonds making substrate more likely to react

COMPETITIVE INHIBITION
Competitive inhibition is reversible. Competitive inhibition is the situation when an inhibiting molecule that is structurally similar to the substrate molecule binds to the active site, preventing substrate binding. eg inhibition of butanedioic acid (succinate) dehydrogenase by propanedioic acid (malonate) in the Krebs cycle

NON-COMPETITIVE INHIBITION
Non-competitive inhibition is the situation where the inhibitor binds to an enzyme (not to its active site) that causes a conformational change in its active site, resulting in a decrease in activity. eg CN inhibition of cytochrome oxidase by binding to SH groups

Characteristic

Competitive inhibition

Non-competitive

COMPARISONstructurally / chemically very inhibitor:

similar to substrate active site blocks active site site of binding: effect:

different from substrate binds to different site / not active site / allosteric site changes 3 structure of enzyme / conformational change of active site substrate cannot bind / reaction not catalyzed / decreased enzyme activity metal ions / Hg+ / Ag+ / Cu2+ / CN inhibit enzymes (cytocrome oxidase) by breaking disulfide linkages

effect: example:

competes with substrate / prevents substrate binding Butanedioic acid (succinate) dehydrogenase by propanedioic (malonate) acid in the Krebs cycle

effect of substrate concentration:

can be reduced by increasing substrate concentration

increasing substrate concentration does not reduce effect of inhibitor

ALLOSTERY

Most allosteric enzymes have multiple allosteric sites. Allosteric inhibition is a form of noncompetitive inhibition. The shape of an allosteric enzyme alternates between active and inactive form.

ALLOSTERIC CONTROL BY END PRODUCT INHIBITION

End-product can inhibit enzyme needed for early / first step in metabolic pathway by changing the shape of the enzyme. This results in negative feedback since increased level of product decreases rate of its own production. Metabolites can act as allosteric inhibitors of enzymes earlier in a metabolic pathway to regulate metabolism. A metabolic pathway is regulated according to the requirement for its end-product.

Substrate
E1

Intermediate 1 End product inhibits first enzyme in the chain thereby inhibiting its own production at high concentrations
E2

E=Enzyme

Intermediate 2
E3

Intermediate 3
E4

Product