الانزيمات د.مصطفى طه محمد Enzymes
- 1. Enzymes Assistant professor Dr. Mustafa Taha Mohammed
- 2. Enzymes Enzyme Action Factors Affecting Enzyme Action Enzyme Inhibition 2
- 3. ENZYMES A protein with catalytic properties due to its power of specific activation
- 4. Enzyme structure • Enzymes are proteins • They have a globular shape • A complex 3-D structure Human pancreatic amylase
- 5. What Are Enzymes? • Most enzymes are Proteins (tertiary and quaternary structures) • Act as Catalyst to accelerates a reaction • Not permanently changed in the process 5
- 6. Enzymes• An enzyme is a biological catalyst • The pockets formed by tertiary and quaternary structure can hold specific substances (SUBSTRATES). • These pockets are called ACTIVE SITES. • When all the proper substrates are nestled in a particular enzyme's active sites, the enzyme can cause them to react quickly • Once the reaction is complete, the enzyme releases the finished products and goes back to work on more substrate.
- 7. What is an enzyme? • Almost all enzymes are proteins that act as biological catalysts. • A catalyst speeds up chemical reactions. Enzymes speed up biological chemical reactions. • Enzymes are highly specific to a type of reaction. • Enzymes must maintain their specific shape in order to function. Any alteration in the primary, secondary, tertiary, or quaternary forms of the enzyme are detrimental.
- 8. Function of enzymes Enzymes have many jobs. They: • Break down nutrients into useable molecules. • Store and release energy (ATP). • Create larger molecules from smaller ones ) • Coordinate biological reactions between different systems in an organism. )
- 9. Enzymes • Catalysts for biological reactions • Most are proteins • Lower the activation energy • Increase the rate of reaction • Activity lost if denatured • May be simple proteins • May contain cofactors such as metal ions or organic (vitamins) 9
- 10. Enzyme Catalyzed Reactions • When a substrate (S) fits properly in an active site, an enzyme-substrate (ES) complex is formed: E + S ES • Within the active site of the ES complex, the reaction occurs to convert substrate to product (P): ES E + P • The products are then released, allowing another substrate molecule to bind the enzyme - this cycle can be repeated millions (or even more) times per minute • The overall reaction for the conversion of substrate to product can be written as follows: E + S ES E + P
- 11. Enzymes • Are specific for what they will catalyze • Are Reusable • End in –ase -Sucrase -Lactase -Maltase 11
- 12. A E B 12
- 13. How do enzymes Work? Enzymes work by weakening bonds which lowers activation energy 13
- 14. Enzymes 14 Free Energy Progress of the reaction Reactants Products Free energy of activation Without Enzyme With Enzyme
- 15. Reaction pathway
- 16. HOW ENZYMES WORK • Enzymes are ORGANIC CATALYSTS. A CATALYST is anything that speeds up a chemical reaction that is occurring slowly. How does a catalyst work? • The explanation of what happens lies in the fact that most chemical reactions that RELEASE ENERGY (exothermic reactions) require an INPUT of some energy to get them going. The initial input of energy is called the ACTIVATION ENERGY
- 17. An enzyme controlled pathway • Enzyme controlled reactions proceed 108 to 1011 times faster than corresponding non-enzymic reactions.
- 18. The substrate • The substrate of an enzyme are the reactants that are activated by the enzyme • Enzymes are specific to their substrates • The specificity is determined by the active site
- 19. Active Site • A restricted region of an enzyme molecule which binds to the substrate. 19 EnzymeSubstrate Active Site
- 20. Enzyme-Substrate Complex The substance (reactant) an enzyme acts on is the substrate 20 EnzymeSubstrate Joins
- 21. Making reactions go faster • Increasing the temperature make molecules move faster • Biological systems are very sensitive to temperature changes. • Enzymes can increase the rate of reactions without increasing the temperature. • They do this by lowering the activation energy. • They create a new reaction pathway “a short cut”
- 22. Chemical reactions • Chemical reactions need an initial input of energy = THE ACTIVATION ENERGY • During this part of the reaction the molecules are said to be in a transition state.
- 23. Enzymes as Biological Catalysts • Enzymes are proteins that increase the rate of reaction by lowering the energy of activation • They catalyze nearly all the chemical reactions taking place in the cells of the body • Enzymes have unique three- dimensional shapes that fit the shapes of reactants (substrates)
- 24. Enzyme Activity The properties of enzymes related to their tertiary structure.The effects of change in temperature,pH,substrate concentration,and competitive and non- competitive inhibition on the rate of enzyme action
- 25. The substrate • The substrate of an enzyme are the reactants that are activated by the enzyme • Enzymes are specific to their substrates • The specificity is determined by the active site
- 26. The active site • One part of an enzyme, the active site, is particularly important • The shape and the chemical environment inside the active site permits a chemical reaction to proceed more easily
- 27. Making reactions go faster • Increasing the temperature make molecules move faster • Biological systems are very sensitive to temperature changes. • Enzymes can increase the rate of reactions without increasing the temperature. • They do this by lowering the activation energy. • They create a new reaction pathway “a ”
- 28. What Affects Enzyme Activity? • Three factors: 1. Environmental Conditions 2. Cofactors and Coenzymes 3. Enzyme Inhibitors 28
- 29. Classification of Enzymes • Enzymes are classified according to the type of reaction they catalyze: Class Reactions catalyzed Oxidoreductases Oxidation-reduction Transferases Transfer groups of atoms Hydrolases Hydrolysis Lyases Add atoms/remove atoms to/from a double bond Isomerases Rearrange atoms Ligases Use ATP to combine molecules
- 30. Examples of Classification of Enzymes • Oxidoreductoases oxidases - oxidize ,reductases – reduce • Transferases transaminases – transfer amino groups kinases – transfer phosphate groups • Hydrolases proteases - hydrolyze peptide bonds lipases – hydrolyze lipid ester bonds • Lyases carboxylases – add CO2 hydrolases – add H2O 30
- 31. Learning Check E1 Match the type of reaction with the enzymes: (1) aminase (2) dehydrogenase (3) Isomerase (4) synthetase A. Converts a cis-fatty acid to trans. B. Removes 2 H atoms to form double bond C. Combine two molecules using ATP D. Adds NH3 31
- 32. Solution E1 Match the type of reaction with the enzymes: (1) aminase (2) dehydrogenase (3) Isomerase (4) synthetase A. 3 Converts a cis-fatty acid to trans. B. 2 Removes 2 H atoms to form double bond C. 4 Combine two molecules using ATP D. 1 Adds NH3 32
- 33. Name of Enzymes • End in –ase • Identifies a reacting substance sucrase – reacts sucrose lipase - reacts lipid • Describes function of enzyme oxidase – catalyzes oxidation hydrolase – catalyzes hydrolysis • Common names of digestion enzymes still use –in pepsin, trypsin 33
- 34. Cofactors • An additional non- protein molecule that is needed by some enzymes to help the reaction • Tightly bound cofactors are called prosthetic groups • Cofactors that are bound and released easily are called coenzymes • Many vitamins are coenzymes Nitrogenase enzyme with Fe, Mo and ADP cofactors ) ©
- 35. Enzyme cofactors cont. • An enzyme that is bonded to its cofactor is called a holoenzyme. • An enzyme that requires a cofactor, but is not bonded to the cofactor is called an apoenzyme. Apoenzymes are not active until they are complexed with the appropriate cofactor.
- 36. Enzyme cofactors • A cofactor is a substance that is not a protein that must bind to the enzyme in order for the enzyme to work. • • A cofactor can be of organic origin. An organic cofactor is called a coenzyme. • • Cofactors are not permanently bonded. Permanently bonded cofactors are called prosthetic groups.
- 37. Enzyme action overview • Enzymes are large molecules that have a small section dedicated to a specific reaction. This section is called the active site. • • The active site reacts with the desired substance, called the substrate • The substrate may need an environment different from the mostly neutral environment of the cell in order to react. Thus, the active site can be more acidic or basic, or provide opportunities for different types of bonding to occur, depending on what type of side chains are present on the amino acids of the active
- 38. Enzyme action theories • Lock and Key: This theory, postulated by Emil Fischer in 1894, proposed that an enzyme is “structurally complementary to their substrates” and thus fit together perfectly like a lock and key. This theory formed the basis of most of the ideas of how enzymes work, but is not completely correct. .,
- 39. Lock-and-Key Model • In the lock-and-key model of enzyme action: - the active site has a rigid shape - only substrates with the matching shape can fit - the substrate is a key that fits the lock of the active site • This is an older model, however, and does not work for all enzymes
- 40. Enzyme Action: Lock and Key Model • An enzyme binds a substrate in a region called the active site • Only certain substrates can fit the active site • Amino acid R groups in the active site help substrate bind • Enzyme-substrate complex forms • Substrate reacts to form product • Product is released 40
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- 42. Lock and Key Model + + E + S ES complex E + P 42 S P P S
- 43. The Lock and Key Hypothesis • Fit between the substrate and the active site of the enzyme is exact • Like a key fits into a lock very precisely • The key is analogous to the enzyme and the substrate analogous to the lock. • Temporary structure called the enzyme-substrate complex formed • Products have a different shape from the substrate • Once formed, they are released from the active site • Leaving it free to become attached to another substrate
- 44. The Lock and Key Hypothesis Enzyme may be used again Enzym e- substr ate compl ex E S P E E P Reaction coordinate
- 45. Enzyme Action: Induced Fit Model • Enzyme structure flexible, not rigid • Enzyme and active site adjust shape to bind substrate • Increases range of substrate specificity • Shape changes also improve catalysis during reaction 45
- 46. Induced Fit • A change in the shape of an enzyme’s active site • Induced by the substrate 46
- 47. Induced Fit • A change in the configuration of an enzyme’s active site (H+ and ionic bonds are involved). • Induced by the substrate. 47 Enzyme Active Site substrate induced fit
- 48. Enzyme Action: Induced Fit Model 48 E + S ES complex E + P S P P SS
- 49. Induced Fit Model • In the induced-fit model of enzyme action: - the active site is flexible, not rigid - the shapes of the enzyme, active site, and substrate adjust to maximumize the fit, which improves catalysis - there is a greater range of substrate specificity • This model is more consistent with a wider range of enzymes
- 50. Learning Check E2 A. The active site is (1) the enzyme (2) a section of the enzyme (3) the substrate B. In the induced fit model, the shape of the enzyme when substrate binds (1) Stays the same (2) adapts to the shape of the substrate 50
- 51. Solution E2 A. The active site is (2) a section of the enzyme B. In the induced fit model, the shape of the enzyme when substrate binds (2) adapts to the shape of the substrate 51
- 52. 2. Cofactors and Coenzymes • Inorganic substances (zinc, iron) and vitamins (respectively) are sometimes need for proper enzymatic activity. • Example: Iron must be present in the quaternary structure - hemoglobin in order for it to pick up oxygen. 52
- 53. Coenzyme reactions • Coenzymes help transfer a functional group to a molecule. • For example, coenzyme A (CoA) is converted to acetyl-CoA in the mitochondria using pyruvate and NAD • Acetyl-CoA can then be used to transfer an acetyl group (CH3CO) to aid in fatty acid synthesis.
- 54. 1. Environmental Conditions 1. Extreme Temperature are the most dangerous - high temps may denature (unfold) the enzyme. 2.pH (most like 6 - 8 pH near neutral) 3.Ionic concentration (salt ions) 54
- 55. Factors that affect enzyme action Enzymes are mostly affected by changes in temperature and pH. • Too high of a temperature will denature the protein components, rendering the enzyme useless. • pH ranges outside of the optimal range will protonate or deprotonate the side chains of the amino acids involved in the enzyme’s function which may make them incapable of catalyzing a reaction.
- 56. Factors Affecting Enzyme Action: Temperature • Little activity at low temperature • Rate increases with temperature • Most active at optimum temperatures (usually 37°C in humans) • Activity lost with denaturation at high temperatures 56
- 57. The effect of temperature • For most enzymes the optimum temperature is about 30°C • Many are a lot lower, cold water fish will die at 30°C because their enzymes denature • A few bacteria have enzymes that can withstand very high temperatures up to 100°C • Most enzymes however are fully denatured at 70°C
- 58. Factors Affecting Enzyme Action Optimum temperature Reaction Rate Low High Temperature 58
- 59. Temperature and Enzyme Activity • Enzymes are most active at an optimum temperature (usually 37°C in humans) • They show little activity at low temperatures • Activity is lost at high temperatures as denaturation occurs
- 60. Factors Affecting Enzyme Action: Substrate Concentration • Increasing substrate concentration increases the rate of reaction (enzyme concentration is constant) • Maximum activity reached when all of enzyme combines with substrate 60
- 61. Substrate concentration: Non-enzymic reactions • The increase in velocity is proportional to the substrate concentration Reactio n velocity Substrate concentration
- 62. Substrate concentration: Enzymic reactions • Faster reaction but it reaches a saturation point when all the enzyme molecules are occupied. • If you alter the concentration of the enzyme then Vmax will change too. Reaction velocity Substrate concentration Vmax
- 63. Substrate Concentration and Reaction Rate • The rate of reaction increases as substrate concentration increases (at constant enzyme concentration) • Maximum activity occurs when the enzyme is saturated (when all enzymes are binding substrate) • The relationship between reaction rate and substrate concentration is exponential, and asymptotes (levels off) when the enzyme is saturated
- 64. Factors Affecting Enzyme Action Maximum activity Reaction Rate substrate concentration 64
- 65. Factors Affecting Enzyme Action: pH • Maximum activity at optimum pH • R groups of amino acids have proper charge • Tertiary structure of enzyme is correct • Narrow range of activity • Most lose activity in low or high pH 65
- 66. Factors Affecting Enzyme Action Reaction Rate Optimum pH 3 5 7 9 11 pH 66
- 67. pH and Enzyme Activity • Enzymes are most active at optimum pH • Amino acids with acidic or basic side-chains have the proper charges when the pH is optimum • Activity is lost at low or high pH as tertiary structure is disrupted
- 68. Enzyme Concentration and Reaction Rate • The rate of reaction increases as enzyme concentration increases (at constant substrate concentration) • At higher enzyme concentrations, more enzymes are available to catalyze the reaction (more reactions at once) • There is a linear relationship between reaction rate and enzyme concentration (at constant substrate concentration)
- 69. Factors that affect enzyme action • Enzymes that can be activated will be affected by the amount of activator or inhibitor attached to its allosteric site. An abundance of an allosteric activator will convert more enzymes to the active form creating more product. • Enzymes that are part of a metabolic pathway may be inhibited by the very product they create. This is called feedback inhibition. The amount of product generated will dictate the number of enzymes used or activated in that specific process.
- 70. Factors that affect enzyme action Enzymes are also affected by the concentration of substrate, cofactors and inhibitors, as well as allosteric regulation and feedback inhibition. (Campbell & Reece, 2002, pp. 99-102) • The concentration of substrate will dictate how many enzymes can react. Too much substrate will slow the process until more enzyme can be made. • The availability of cofactors also dictate enzyme action. Too little cofactors will slow enzyme action until more cofactors are added. • An influx of competitive or non-competitive inhibitors will not necessarily slow the enzyme process, but will slow the amount of desired product.
- 71. Learning Check E3 Sucrase has an optimum temperature of 37°C and an optimum pH of 6.2. Determine the effect of the following on its rate of reaction (1) no change (2) increase (3) decrease A. Increasing the concentration of sucrose B. Changing the pH to 4 C. Running the reaction at 70°C 71
- 72. Solution E3 Sucrase has an optimum temperature of 37°C and an optimum pH of 6.2. Determine the effect of the following on its rate of reaction (1) no change (2) increase (3) decrease A. 2, 1 Increasing the concentration of sucrose B. 3 Changing the pH to 4 C. 3 Running the reaction at 70°C 72
- 73. Enzyme Inhibition Inhibitors • cause a loss of catalytic activity • Change the protein structure of an enzyme • May be competitive or noncompetitive • Some effects are irreversible 73
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- 75. Two examples of Enzyme Inhibitors a. Competitive inhibitors: are chemicals that resemble an enzyme’s normal substrate and compete with it for the active site. 75 Enzyme Competitive inhibitor Substrate
- 76. Inhibitors 76 b. Noncompetitive inhibitors: Inhibitors that do not enter the active site, but bind to another part of the enzyme causing the enzyme to change its shape, which in turn alters the active site. Enzyme active site altered Noncompetitive Inhibitor Substrate
- 77. Enzyme Inhibitors • Inhibitors (I) are molecules that cause a loss of enzyme activity • They prevent substrates from fitting into the active site of the enzyme: E + S ES E + P E + I EI no P formed
- 78. Competitive Inhibition A competitive inhibitor • Has a structure similar to substrate • Occupies active site • Competes with substrate for active site • Has effect reversed by increasing substrate concentration 78
- 79. Reversible Inhibitors (Competitive Inhibition) • A reversible inhibitor goes on and off, allowing the enzyme to regain activity when the inhibitor leaves • A competitive inhibitor is reversible and has a structure like the substrate - it competes with the substrate for the active site - its effect is reversed by increasing substrate concentration
- 80. Noncompetitive Inhibition A noncompetitive inhibitor • Does not have a structure like substrate • Binds to the enzyme but not active site • Changes the shape of enzyme and active site • Substrate cannot fit altered active site • No reaction occurs • Effect is not reversed by adding substrate 80
- 81. Reversible Inhibitors (Noncompetitive Inhibition) • A noncompetitive inhibitor has a structure that is different than that of the substrate - it binds to an allosteric site rather than to the active site - it distorts the shape of the enzyme, which alters the shape of the active site and prevents the binding of the substrate • The effect can not be reversed by adding more
- 82. Learning Check E4 Identify each statement as describing an inhibitor that is (1) Competitive (2) Noncompetitive A. Increasing substrate reverses inhibition B. Binds to enzyme, not active site C. Structure is similar to substrate D. Inhibition is not reversed with substrate 82
- 83. Solution E4 Identify each statement as describing an inhibitor that is (1) Competitive (2) Noncompetitive A. 1 Increasing substrate reverses inhibition B. 2 Binds to enzyme, not active site C. 1 Structure is similar to substrate D. 2 Inhibition is not reversed with substrate 83
- 84. The switch: Allosteric inhibition Allosteric means “other site” E Active site Allosteric site
- 85. End point inhibition • The first step (controlled by eA) is often controlled by the end product (F) • Therefore negative feedback is possible A B C D E F • The end products are controlling their own rate of production • There is no build up of intermediates (B, C, D and E) eFeDeCeA eB Inhibition
- 86. The allosteric site the enzyme “on-off” switch E Active site Allosteri c site empty Substrat e fits into the active site The inhibitor molecule is absent Conformational change Inhibitor fits into allosteric site Substrate cannot fit into the active site Inhibitor molecule is present E
- 87. Switching off • These enzymes have two receptor sites • One site fits the substrate like other enzymes • The other site fits an inhibitor molecule Inhibitor fits into allosteric site Substrate cannot fit into the active site Inhibitor molecule
- 88. Isoenzymes • Isoenzymes are different forms of an enzyme that catalyze the same reaction in different tissues in the body - they have slight variations in the amino acid sequences of the subunits of their quaternary structure • For example, lactate dehydrogenase (LDH), which converts lactate to pyruvate, consists of five isoenzymes
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- 90. الصغائكم ارشك بوك الفيس:د.محمد طه مصطفى االلكتروني البريد:[email protected]