ENZYMES

Define the following terms:1. Anabolic reactions:

2. Catabolic reactions:

3. Metabolism:

4. Catalyst:

5. Metabolic pathway:

6. Specificity:

7. Substrate:

8. Product:

Reactions that build up molecules

Reactions that break down molecules

Combination of anabolic and catabolic reactions

Sequence of enzyme controlled reactions

Only able to catalyse specific reactions

The molecule(s) the enzyme works on

Molecule(s) produced by enzymes

A substance that speeds up reactions without changing the produced substances

Naming enzymes:

• Intracellular enzymes

• Extracellular enzymes

• Recommended names

• Systematic name

• Classification number

Work inside cells eg.DNA polymerase

Secreted by cells and work outside cells eg. pepsin, amylase

Short name, often ending in ‘ase’ eg. creatine kinase

Describes the type of reaction being catalysed eg.ATP:creatine phosphotransferase

Eg. 2.7.3.2

Timeline of enzyme discovery

1835:Breakdown of starch to sugar by malt

1877:Name enzyme coined to describe chemicals in yeast that ferment sugars

1897:Eduard Buchner extracted enzyme from yeast and showed it could work outside cells

1926:James B Sumner produced first pure crystalline enzyme (urease)

and showed enzymes were proteins

1905:Otto Rohm exyracted pancreatic proteases to supply enzymes for tanning

1930-1936:Protein nature of enzymes finally established when digestive enzymes

crystallised by John H Northrop

1946: Sumner finally awarded Nobel prize

Enzymes lower the activation energy of a reaction

Final energy state of products

Initial energy stateof substrates

Activation energyof uncatalysed reactionsActivation energy

of enzyme catalysedreaction

Progress of reaction (time)

Ene

rgy

leve

ls o

f m

olec

ules

Enzymes lower activation energy by forming an enzyme/substrate complex

Substrate + Enzyme

Enzyme/substrate complex

Enzyme/product complex

Product + Enzyme

In anabolic reactions enzymes bring the substrate molecules together.

In catabolic reactions the enzyme active site affects the bonds in substrates so they are easier to break

Lock-and-key hypothesis assumes the active site of an enzyme is rigid in its shape

How ever crystallographic studies indicate proteins are flexible.

The Induced-fit hypothesis suggests the active site is flexible and only assumes its catalytic conformation after the substrate molecules bind to the site.

When the product leaves the enzyme the active site reverts to its inactive state.

Enzymes are globular proteins

• Active site has a specific shape due to tertiary structure of protein.

• A change in shape of the protein affects shape of active site and the function of the enzyme.

Click to link to jmol interactive representation courtesy of University of Arizona

Characteristics of enzymes

• Only change the rate of reaction. They do not change the equilibrium or end products.

• Specific to one particular reaction

• Present in very small amounts due to high molecular activity:

Turnover number = number of substrate molecules transformed per minute by one enzyme molecule

Catalase turnover number = 6 x106/min

How would you measure the effect of an enzyme?

• Compare uncatalysed rate with catalysed.

• Enzymes can increase rate by a factor of between 108 to 1026

Characteristics of enzymes

• Rate of enzyme action is dependent on number of substrate molecules present

Vmax = maximum rate of reaction

Vmax approached as all active sites become filled

Some active sites free at lower substrate concentrations

Substrate concentration

Rat

e of

Rea

ctio

n (M

)

Why do scientists measure the initial rate of reaction of enzyme-catalysed reactions?

Rat

e of

Rea

ctio

n (M

)

Independent variable

Initial rate of reaction

They measure rate at start of reaction before any factors, eg. substrate concentration, have had time to change.

Rate of enzyme –catalysed reactions are affected by temperature.

Temperature coefficient Q10:

rate of reaction at (x + 10) oC

Q10 = -----------------------------------------

rate of reaction at x oC

Q10 for between 0 - 40 oC is 2

Enzymes denature at 60oC

Temperature

Rat

e of

rea

ctio

n

Rate doubles every 10oC

Enzyme denaturing and losing catalytic abilities

Optimum temperature

Some thermophilic bacteria have enzymes with optimum temperatures of 85oC

pH affects the formation of hydrogen bonds and sulphur bridges in proteins and so affects shape.

pepsintrypsin cholinesterase

2 4 8 106pH

Rat

e of

Rea

ctio

n (M

)

Enzymes in medicine

Glucose oxidaseGlucose Hydrogen peroxide

peroxidase

Dye: Blue---Green---Brown

Dye changes according to amount of glucose

Enzyme-linked immunosorbent assays (ELISAs) detect antibodies to infections.

Glucose oxidase + peroxidase + blue dye on dipsticks to detect glucose in urine: