ENZYMES

Biological Catalysts
Presentation by Muhammad Bilal
EasyLearningHome.com
ENZYMES
▪ Most important group of proteins
▪ Biological catalysis
▪ Increase efficiency of reactions
• Too slow without enzymes
• Life impossible
▪ Specific for each reaction
▪ Polypeptide chains
• Coiled in globular conformation
ACTIVE SITE
▪ Active site
• Small portion of enzyme
• Has catalytic activity
• Only few amino acids
▪ Other amino acids maintain structure
▪ Substrate
• Substance on which enzyme acts
COFACTOR
▪ Some enzymes are protein only
▪ Others need non-protein part
▪ Called cofactor
▪ Essential for proper function
• Sometimes acts as bridge
• Sometimes contributes in reaction
• Sometimes provide energy
COFACTOR
▪ Three types
▪ Activators
• Heavy-metal cofactors
• detachable
• Mg+2, Fe+2, Cu+2, Zn+2
▪ Prosthetic Group
• Covalently bonded to protein part
• E.g. Heme of hemoglobin
▪ Coenzyme
• Loosely attached
• Related to vitamins
• E.g. NAD
APOENZYME & HOLOENZYME
▪ Apoenzyme
• Enzyme without Coenzyme / prosthetic group
• Needs coenzyme for activation
▪ Holoenzyme
• Enzyme and cofactor
• Activated
ENZYMES
▪ Some are dissolved in cytoplasm
▪ Some bound to certain organelles
• Produced at the site of use
▪ Enzymes for photosynthesis
• Present in chloroplast
▪ Enzymes for respiration
• Present in mitochondria
▪ Enzymes for protein synthesis
• Present in ribosomes
CHARACTERISTICS OF ENZYMES
▪ Biological catalysts
▪ Globular proteins
▪ Increase rate of reaction
• Without being used up
• Lower activation energy
▪ Nature of products in not changed
▪ Small amount is enough
▪ Specific action
• Special enzyme for each reaction / related reactions
▪ Sensitive to pH, temperature, substrate concentration
▪ Some need cofactor for functioning
ACTIVATION OF ENZYMES
▪ Some enzymes have damaging effects
▪ Produced in inactive form
▪ E.g. Pepsin
• Produced as pepsinogen
• Remains inactive inside cell
• Released in stomach
• Activated by acid
• Deactivated after digestion
MECHANISM OF ENZYME ACTION
▪ Globular protein
▪ Specific chemical composition
• Specific amino acid sequence
• Specific three dimensional shape
▪ Recognizes a specific substance
• Called substrate
• Binds to it
• Transforms into product
▪ Released and can be used again
MECHANISM OF ENZYME ACTION
▪ Enzyme and substrate react
• Enzyme substrate complex formed
▪ Active site
• Special charge bearing site
• Charge and shape due to amino acids
• Coiled and folded
▪ Has two portions
▪ Binding site
• Recognizes substrate
▪ Catalytic site
• Transforms substrate into product
LOCK AND KEY MODEL
▪ Emil Fisher (1890)
▪ Describes substrate enzyme interaction
▪ Specific enzyme can convert only specific substrate
▪ Like each lock has a specific key
▪ Supposes enzyme has rigid structure
• No flexibility
• No change in shape
• Before, during, or after conversion
▪ Experimental results rejected it
INDUCED FIT MODEL
▪ Koshland (1959)
• On the basis of new data
▪ Substrates combines with enzyme
▪ Induces change in shape
▪ Enables enzyme to perform catalysis
▪ Suggests enzyme has flexible shape
▪ More acceptable
BIOCHEMICAL PATHWAYS
▪ Some enzymes work together
▪ Series of chemical reactions
▪ Product of one enzyme – substrate of next
▪ Enzymes present in precise order
• Substrate moves from one to next
▪ Converted in small steps
▪ Until final product
▪ E.g. Photosynthesis, glycolysis
NEGATIVE FEEDBACK
▪ Inhibition
▪ In Biochemical pathways
▪ Final product accumulates
▪ Some molecules block first enzyme
▪ Reaction is inhibited
▪ No more product forms
POSITIVE FEEDBACK
▪ Precursor Activation
▪ Large amount of substrate
▪ Sometimes binds to enzymes
▪ Activates them
▪ To promote conversion into product
▪ Substrate gradually decreases
FACTORS AFFECTING ENZYMES
▪ Function depends on structure
▪ Globular Proteins
• Highly sensitive
▪ Affected by various factors
• Factors that affect structure
▪ 1. Enzyme Concentration
▪ 2. Substrate Concentration
▪ 3. Temperature
▪ 4. pH
▪ 5. Inhibitors
ENZYME CONCENTRATION
▪ Rate depends on amount of enzyme
▪ More enzyme
• More active sites
• More substrate binds
• Rapid conversion to products
▪ When substrate is unlimited
• Doubling enzyme double rate of reaction
▪ At very high concentration
• Called limiting concentration
• Rate no longer depends on it
SUBSTRATE CONCENTRATION
▪ At low concentration
• Abundant enzyme
• Rate depends on amount of substrate
• Increasing substrate increases rate
▪ At a certain point
• All active sites occupied
• Rate no longer increases
• With increasing substrate
TEMPERATURE
▪ Heat provides kinetic energy
• Collision of substrate and enzyme
▪ Heat provides activation energy
• Helps quickly start reaction
▪ Rate increases with temperature
• Up to certain limit
▪ Optimum temperature
• Working at maximum rate
• Specific for each enzyme (usually 37°C)
▪ Too high temperature
• Too much vibration in molecules
• Breaks hydrogen bonds
• Destroys globular structure - denatured
PH VALUE
▪ Optimum pH
• Narrow range of pH
• Maximum rate of reaction
▪ Change in pH
• Add or remove H+
• Disturbs charged groups of amino acids
• Enzyme activity is disturbed
• Activity retarded or blocked
▪ Extreme change in pH
• Hydrogen bonds break
• Denaturation of enzyme
INHIBITORS
▪ Substance that reacts with enzyme
• Not transformed into product
• Blocks activity of enzyme
• Temporarily or permanently
▪ Examples
• Poisons – block important enzymes
• Antibiotics – block enzymes of bacteria
• Drugs – block enzymes producing “pain”
▪ Two types:
• Irreversible inhibitors
• Reversible inhibitors
INHIBITORS
▪ Irreversible inhibitors
• Usually form covalent bonds
• Permanently occupy active site
• Or destroy shape of enzyme
• Reaction is blocked
▪ Reversible inhibitors
• Form weak linkage with enzyme
• Effects can be neutralized
• By increasing substrate concentration
• Two types
• Competitive inhibitors
• Non-competitive inhibitors
INHIBITORS
▪ Competitive Inhibitors
• Structure similar to substrate
• Selected by binding site
• Not converted by catalytic site
• No products formed
▪ Non-competitive inhibitors
• Form complex with enzyme
• Site different than active site
• Change structure of enzyme
• Substrate cannot bind
• No product formed