1. E N Z Y M E SE N Z Y M E S

2. Enzyme : – mostly proteins, but some catalytic RNA molecules (ribosymes) – extraordinary catalytic power – high degree of specificity for their substrate – tremendously accelerate chemical reactions – function in aqueous solutions under very mild conditions of temperature and pH – can be regulated

3. History of enzyme research: –late 1700s biological catalysis recognized and described –early 1800s conversion of starch to sugar by saliva –1850 Louis Pasteur fermentation of sugar into alcohol by yeast is catalysed by “ferments” –1897 Eduard Buchner yeast extractum- can work without living cell- later Frederick Kühne called these molecules ENZYMES –1926 James Sumner- urease he postulated that “all enzymes are proteins” –1930s John Northrop and Moses Kunitz crystallized trypsin, pepsin and other digestive enzymes –JBS Haldane’s treatise entitled “Enzymes” –Since the later part of the twentieth century intensive research on enzymes- purification of thousands of enzymes and elucidation of the structure and chemical mechanism of them

4. E. C. (Enzyme Commission) number enzymes are classified by the reactions they catalyze EC 1. Oxidoreductases These enzymes catalyze oxidation and reduction reactions. - dehydrogenases act on the following functional groups as e - donors: -CH 2 =CH 2 -, -CH 2 -NH 2 -, -CH=NH, NADH and NADPH - oxidases2 e - transfer to O 2 - usually H 2 O 2 formation - oxygenasescatalyze incorporation of both atoms of O 2 into the substrate - hydroxylasesincorporation of one atom of O 2 (steroid hydroxylases) - peroxidasesutilize H 2 O 2 rather than O 2 as the oxidant - catalaseunique in that, H 2 O 2 serves as both donor and acceptor. Function in the cell to detoxify H 2 O 2 H 2 O 2 + H 2 O 2 2 H 2 O + O 2

5. EC 2. Transferases They are involved in transferring functional groups between donors and acceptors- transferred amino, acyl, phosphate one-carbon and glycosyl groups. - aminotransferases- transfer amino group to a keto acid acceptor - kinases- transfer phosphoryl group to alcohol or amino group acceptors EC 3.Hydrolases They catalyze hydrolysis of different bonds. The donor group is transferred to water. The cleavage of the peptide bond is a good example of this reaction.

6. EC 4.Lyases They add or remove the elements of H 2 O, NH 3 or CO 2. - decarboxylases - aldolases -hydratases - dehydratases -synthases EC 5. Isomerases They catalyze isomerization of several types. - racemases, epimerases- inversion at asymmetric carbons - isomerases (cis-trans) - intramolecular transferases - mutases- intramolecular transfer of a group

7. EC 6. Ligases These enzymes are involved in synthetic reactions where two molecules are joined. - synthetases - carboxylases

8. –proteins or ribosymes –integrity the native protein conformation –molecular weight 12000- 1 million D –require no chemical group for activity –require COFACTOR –require COENZYME –PROSTHETIC group Holoenzyme:complete, catalytically active enzyme together with its bound coenzyme and/or metal ions. Apoenzyme (apoprotein): protein part of such an enzyme

11. –catalytic power –specificity for their substrate –accelerate chemical reactions –transform different energy forms –can be regulated Generally about enzymes

14. Enzymes influence the reaction rate, but they do not affect reaction equilibria! Enzymes enhance reaction rate by: stabilizing the transition state lowering activation energies

15. –entropy reduction (restriction in the relative motions of the substrates that are to react) –desolvation of the substrate –change in conformation of the substrate –exact fit of enzyme and substarte Binding energy

16. –catalytic group (break or form different binds) –small size (enzyme itself is large) –three dimensional structure –binding substrate with multiple weak interactions – this gives specificity Active center of enzymes

18. Specific catalytic groups contribute to catalysis Acid-base catalysis specific general Covalent catalysis Metal ion catalysis

19. Enzyme kinetics

20. Simple enzymatic reaction: E + SESEPE + P v 0 = v max [S] K M + [S]

26. Enzyme inhibition

30. –Methanol or ethylene-glycol poisoning Th: ethanol infusion –sulfa drugs (sulfanilamide) - antibiotics - competes with p-aminobenzoic acid, which is required for bacterial growth –Methotrexate - biosynthesis of DNA and RNA requires folic acid, and its structural analog- chemotherapic agent Medical significance of competitive inhibition:

44. –enzymes with different molecular structure, catalyzing the same biochemical reactions –they are either the products of different genes or complexes of a few subunit types, mixed at different ratios –heterogenity in their K M value (- different substrate affinity) –represented in different tissue, cell types or compartments by their different isoforms Izoenzymes: –ways can be distinguished: ~ electrophoretic separation ~ application of isoform specific antibodies ~ application of isoform specific inhibitors ~ on the basis of different heat sensitivity

45. CPK1BB CPK2MBmyocardium CPK3MMskeletal muscle