ENZYMES

Define the following terms: Anabolic reactions: Catabolic reactions: Metabolism: Catalyst: Metabolic pathway: Specificity: Substrate: Product: Reactions that build up molecules Reactions that break down molecules Combination of anabolic and catabolic reactions A substance that speeds up reactions without changing the produced substances Sequence of enzyme controlled reactions Only able to catalyse specific reactions The molecule(s) the enzyme works on Molecule(s) produced by enzymes

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 1905: Otto Rohm exyracted pancreatic proteases to supply enzymes for tanning 1926: James B Sumner produced first pure crystalline enzyme (urease) and showed enzymes were proteins 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 of uncatalysed reactions Initial energy state of substrates Activation energy of enzyme catalysed reaction Energy levels of molecules Final energy state of products Progress of reaction (time)

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 Rate of Reaction (M) Some active sites free at lower substrate concentrations Substrate concentration

Why do scientists measure the initial rate of reaction of enzyme-catalysed reactions? They measure rate at start of reaction before any factors, eg. substrate concentration, have had time to change. Rate of Reaction (M) Independent variable

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 Optimum temperature Enzyme denaturing and losing catalytic abilities Rate doubles every 10oC Rate of reaction 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. trypsin cholinesterase pepsin Rate of Reaction (M) 2 4 6 8 10 pH

Enzymes in medicine Glucose oxidase + peroxidase + blue dye on dipsticks to detect glucose in urine: Glucose oxidase Glucose Hydrogen peroxide peroxidase Dye: Blue---Green---Brown Dye changes according to amount of glucose Enzyme-linked immunosorbent assays (ELISAs) detect antibodies to infections.