1. Enzymes Classification: a. Intracellular enzymes
Biocatalyst synthesized in the living cells, mostly protein in nature,thermolabile in character and specific in their action.Enzymes are proteins functioning as catalysts that speed up reactions by lowering the activation energy.The enzyme catalysts regulate the fuction of cells and organisim,
Classification: a. Intracellular enzymes
b. Extracellular enzymes

17. A four digit enzyme commission(E.C.)
According to international union of biochemistry (IUB) divided into six major groups:
1-Oxidoreducatase Oxidation reduction reaction
2-Transferase catalyze the transfer of functional group
3-Hydrolase hydrolysis of various compound
4-Lyases specialized in addition or removal of
water ,ammonia, CO2.
5-Isomerase all isomerization reaction.
6- Ligases catalyses the synthetic reaction
According to the reaction being catalyzed.
Another classification: that each class in turn is subdivided into many subclass which are further divided.
A four digit enzyme commission(E.C.)

25. Enzymes also classified on the basis of their composition .
enzyme, Enzymes wholly of proteins are known as simple
in contrast to complex enzyme which are
composed of protein+non protein molecule.
Complex enzyme holoenzyme.
Protein component in complex enzyme apoenzyme.
Non protein component is called coenzyme or prosthetic
group

26. Structure of enzymes Enzymes
Complex or holo_enzymes (protein part and non protein part – cofactor)
Simple (only protein)
Apoenzyme (protein part)
Non protein part Cofactor
Prosthetic groups
usually small inorganic molecule or atom;
usually tightly bound to apoenzyme
Coenzyme
-large organic molecule
-loosely bound to apoenzyme

27. Examples : vitamins or compound derived from vitamins.
Cofactors : responcible for :
a. Bond formation between enzyme and substrate
b. Transfer of functional group.
c. Takes place in the formation of tertiary structure of protein part.
Coenzyme: The non-protein component, loosely bound to apoenzyme by non-covalent bond.
Examples : vitamins or compound derived from vitamins.
Prosthetic group The non-protein component, tightly bound to the apoenzyme by covalent bonds is called a Prosthetic group.

28. Prosthetic group Metal ions
. Heme group of cytochrome.
. Biotin of acetyl-CoA CARBOXYLASE.
Metal ions
Fe –cytochrome oxidase, catalase
Cu – cytochrome oxidase, catalase
Zn – alcohol dehydrogenase
Mg – hexokinase, glucose-6-phosphatase
K, Mg –pyruvate kinase
Na, k –ATP-ase
Coenzyme
TPP, FMN, FAD, pantothenic acid, COA, NAD,NADPH

29. Enzymes that require a metal in their composition known as metalloenzymes
Two group of enzyme requiring metals for their activity are distinguished;
A: metal-activated enzyme: metal is not tightly held by enzyme and can be exchanged easily.EX: ATPase(Mg+2,Ca+2).
B: Metalloenzymes: enzymes hold tightly and not exchangable:Ex: alcohol dehydrogenase,Alkaline phosphatase

30. Factors affecting enzyme activity:
1.Concentration of enzyme.
2. Concentration of substrate.
3.Effect of Temperature.
4.Effect of PH.
5. Effect of product concentration.
6. Effect of activator.

31. 1. Conc. Of enzyme: as the conc
1. Conc. Of enzyme: as the conc. Of enzyme is increased ,the velocity of the reaction is increased.

32. 2 Conc. Of the substrate: increase in the substrate
conc. gradually increase the velocity of enzyme
reaction within the limited range of substrate level.
distinct phases of reaction are observed in the graph:
a/ at a low substrate concentration the velocity of reaction is directly proportional to the substrate level (phase A).
b/In the second phase (phase B) it is not directly proportional to the enzyme activity.

33. c/ In the third phase (phase C ) the reaction is independent of the substrate concentration.

36. The Michaelis-Menten Equation is the Fundamental Equation of Enzyme Kinetics
E + S ES E + P k1 k2
k-1
E = enzyme concentration S = Substrate concentration ES = Enzyme-substrate complex concentration (noncovalent) P = product concentration k1 = rate constant for formation of ES from E + S k-1 = rate constant for decomposition of ES to E + S. k2 = rate constant for decomposition of ES to E + P.

37. Development of the Michaelis-Menton Equation
E + S ES E + P k1 k2
k-1
1. The overall rate of product formation: v = k2 [ES]
2. Rate of formation of [ES]: vf = k1[E][S]
3. Rate of decomposition of [ES]:
vd = k-1[ES] + k2 [ES]
4. The steady state assumption requires that: Rate of ES formation = Rate of ES decomposition
5. So: k1[E][S] = k-1[ES] + k2 [ES]

38. Michaelis-Menton Derivation
6. In solving for [ES], use the enzyme balance to eliminate [E]. ET = [E] + [ES]
7. k1 (ET - [ES])[S] = k-1[ES] + k2 [ES]
k1 ET[S] - k1[ES][S] = k-1[ES] + k2 [ES]
8. Rearrange and combine [ES] terms:
k1 ET[S] = (k-1 + k2 + k1 [S])[ES]
k1 ET[S]
9. Solve for [ES] =
(k-1 + k2 + k1 [S])

39. Michaelis-Menton Derivation
ET[S]
10. Divide through by k1: [ES] =
(k-1 + k2)/k1 + [S]
11. Defined Michaelis constant: KM = (k-1 + k2) / k1
12. Substitute KM into the equation in step 10.
13. Then substitute [ES] into v = k2 [ES] from step1 and replace Vmax with k2 ET to give:
Vmax[S] vo = KM + [S]

43. 3.Effect of Temp: velocity of an enzyme reaction increased with increase in Temp. up to maximum and decline. A bell shape curved is usually observed.

44. The optimum temp. for most of the enzymes is between (40-45)C.
When enzymes are exposed to a Temp above 50C denaturation occurs leading to dearrangment in the tertiary structure of protein.

45. 4. Effect of PH: increase in (H) ion concentration(PH) influence the enzyme activity and bell- shaped curve is normally obtained

46. Each enzyme has own optimum PH at which the velocity maximum.
Most of the enzyme showed optimum activity around neutral PH , PH(6-8)
5-Effect of product concentration: the accumulation of reactive product generally decrease the enzyme velocity .
For certain enzymes , the product combine with active site of the enzyme and form a loose complex and this inhibit the enzyme activity.
6.Effect of activator:
Some enzymes require certain inorganic metallic cation, Mg+2,Mn+2,Zn+2,Ca+2. for their optimum activity, these metals function as activator of the enzyme velocity through various mechanism

47. Enzyme Inhibition:
Enzyme Inhibitor: a substrate which binds with enzyme and bring about a decrease in catalytic activity of that enzyme
Enzyme Inhibitor groups
Reversible Irreversible Allosteric
Enzyme Inhibitors
Inhibitors are chemicals that reduce the rate of enzymic reactions.
They are usually specific and they work at low concentrations.
They block the enzyme but they do not usually destroy it.
Many drugs and poisons are inhibitors of enzymes in the nervous system.

48. Meaning of enzyme inhibitors
Irreversible inhibitors : Inhibition of enzyme activity by combining with active site.
Reversible inhibitors : Inhibitors binds non-covalently with the enzyme and the enzyme can be reversed if the inhibitor is removed.
a-Competitive inhibitors : Inhibition of enzyme activity by competing with active site .
b-Un Competitive inhibitors : Inhibition of enzyme activity by combining with allosteric site.
c-Non Competitive inhibitors : Inhibition of enzyme activity by combining with both to free enzyme and ES at allosteric site.

53. a.Competative inhibition:
The inhibitor is closely resemble the substrate {S} substrate analogue
The inhibitor competes with the substrate and binds at the active site of the enzyme but doesn't undergo any catalysis.
Ex. Of clinical &pharmacological inhibition.
Xanthine oxidase
Hypoxanthine
xanthine
Uric Acid
Allopurinol
Allopurinol acts as competitive inhibitor to xanthine oxidase , it competes with enzyme xanthine oxidase and prevent or block the reaction.
so control Gout ( decrease uric acid production).

54. They are used for cancer therapy
Dihydrofolate
Reductase
Dihydrofolic acid
Tetrahydrofolic acid FH4
Folinic acid
methotrxate
Antimetabolites
are chemical compound that block the reaction by their inhibitor effect on the enzyme .
Antimetabolite are structural analogues of substrate and they are a competitive inhibitor .
They are used for cancer therapy

60. هو غاز الاعصاب اكتشف من قبل الالمان في الحرب العالمية الثانية
2.Irreversable Inhibition:
The inhibitors bind covalently with the enzymes and inactivate them which is irreversible and the inhibition is usually toxic substance.
Ex:Di isopropyl flurophosphate(DFP) is a nerve gas irreversibly bind with enzyme at the active site.
هو غاز الاعصاب اكتشف من قبل الالمان في الحرب العالمية الثانية
Organophosphorous compound(insecticides) block the activity of Ach.esterase essential for nerve conduction resulting in paralysis of vital body function.
Diagnostic importance of Enzymes:
Measurement of enzyme activities in biological fluid (plasma/serum) is of great clinical importance.
Enzymes in the circulation is divided into two groups:
1/ plasma specific or plasma functional enzymes:
Certain enzymes are normally present in the plasma and they have specific function.

61. Generally these enzymes activities are higher in plasma than in tissues. They are mostly synthesized in the liver and enter the circulation.
Ex: lipoprotein lipase plasmin, choline esterase.
Impairment of liver function or genetic disorders leads to fall in the activities of plasma function enzyme.
2/Non –plasma specific: these enzymes are either totally absent or present at a low concentration in plasma compared to their level found in the tissues.
All GIT enzymes ,amylase, pepsin,trypsin present in the plasma are known as secretary enzymes.
Measurement of the activities of non plasma specific is important in the diagnosis and prognosis of several diseases