MCMT 20 - ENZYMES

Prepared by: ABRMT

 Catalysts – substances that speed up the rate of a - Acts with and is essential to enzyme activity
chemical reaction
Types of Cofactors
 Catalysis - The process of increasing the rate of
chemical reactions 1. Coenzyme
 Substrate – substance that binds to the active site - nonprotein organic substance
of enzyme - dialyzable, thermostable
- loosely attached to the protein part
ENZYMES
2. Prosthetic group
- Biological catalysts - organic substance
- Most efficient catalysts known - dialyzable, thermostable
 Can increase the rate of reaction by a factor up - firmly attached to the protein and apoenzyme
to 1020 portion
- Specific for a substrate that it converts to a defined 3. Metal-ion Activator
product
- Enzyme lowers the activation energy of a reaction  Holoenzyme
 ACTIVATION ENERGY - Energy input required to - Apoenzyme + Cofactor
initiate the reaction - Functional unit
- Frequently appear in serum following cellular
ENZYME THEORY
injury / degradation of cells
- Usually globular proteins  Emil Fischer’s / Lock & Key
- Active site has a rigid shape
Enzyme Functions:
- Only substrate with matching shape can fit
1. Catalysis – most important  Kochland’s / Induced Fit
2. Can distinguish stereoisomers of given compound - Active site is flexible
3. Finely tuned by regulatory process - Binding of substrate induces a conformational
change in the enzyme
Parts: - Greater range of substrate specificity
 Active Site CLASSIFICATION OF ENZYMES
- Site where substrate attaches forming E-S
Complex 1. Oxidoreductase
 Allosteric Site - Catalyzed reaction: Redox reaction
- Cavity that bind substances that inhibit or - Examples: Lactate Dehydrogenase, Glucose-6-
stimulate enzyme activity Phosphate Dehydrogenase

Components: Includes:

 Apoenzyme - Dehydrogenase
- Inactive enzyme - Oxidase
- Protein portion which is mainly responsible for - Oxygenase
enzyme action - Reductase
 Cofactor - Peroxidase
- Nonprotein entities that must bind to
particular enzyme 2. Transferase
 MCMT 20 - ENZYMES
Prepared by: ABRMT

- Catalyzed reaction: Transfer of functional group - Acts only on molecules that have specific
(other than hydrogen) functional group
- Examples: Aspartate Aminotransferase, Alanine - Hexokinase adds phosphate group to hexoses
Aminotransferase, Creatine Kinase
3. Linkage Specificity
Includes: - Acts on a particular type of chemical bond
- Transaminase
- Kinase 4. Stereo Chemical Specificity
- Phosphorylase - Acts with only one optical isomer of a certain
compound
3. Hydrolase - L-amino acid oxidase acts only on L-amino acids
- Catalyzed reaction: Hydrolysis of various bonds
ENZYME NOMENCLATURE
by addition of water
- Examples: Alkaline Phosphatase, Amylase 1. Recommended Name
- Named according to the reaction they carry out
Include: - Suffix: -ase
- Digestive enzymes 2. Systematic Name (International Classification)
- Lysozymes - According to the numerical designation gicen by
- Phosphatase the Enzyme Commission (E.C.)
- Nucleases a. First number – enzyme classification
b. Next two numbers – subclass and subsubclass
4. Lyase c. Last number – serial number (specific to each
- Catalyzed reaction: Removal of groups from enzyme)
substrates without hydrolysis and the resulting
6 Functional Classes
product contains double bonds. EC1 Oxidoreductase
- Examples: Aldolase, decarboxylase EC2 Transferase
EC3 Hydolase
5. Isomerase EC4 Lyase
- Catalyzed reaction: Interconversion of EC5 Isomerase
geometric, optical, or positional isomers. EC6 Ligase
- Examples: Glucose phosphate isomerase
FACTORS AFFECTING ENZYME ACTIVITY
6. Ligase
- Catalyzed reaction: Joining of 2 substrate  Temperature
molecules utilizing ATP - ↑temp = ↑rate of chemical rxn = ↑movement
- Examples: Glutathione synthetase of molecules
- Active @ 25C, 30C, or 37C
SPECIFICITY OF ENZYMES
- Denatured @ 40C – 50C
1. Absolute Specificity - Inactivated @ 60C – 65C or if frozen
- Acts on only one substrate and one reaction - Stored @ 5C
2. Group Specificity  pH
- Stability factor of enzyme
 MCMT 20 - ENZYMES
Prepared by: ABRMT

- Most physiologic enzyme reaction occurs at the

pH range of 7.0 to 8.0
- Optimum activity varies
 Enzyme Concentration
- ↑ enzyme level = faster reaction
- Because more enzyme will bind to the substrate
 Substrate Concentration
- If the amount of the enzyme is kept constant
and the substrate concentration is then
gradually increased, the reaction velocity will
increase until it reaches a maximum

 Substrate Saturation - The concentration at

which it reaches its maximum rate and all the
active sites are full
 Turnover Number - Number of substrate
molecules converted to product/second/enzyme
molecule under conditions of optimum
temperature and pH
 Machaelis-Menten Equation – Vmax equation
- Increased in substrate concentration will not
increase the velocity (maximum velocity)
- When reached, all enzymes are already
converted to enzyme-substrate complex

 Inhibitors
1. Competitive Inhibitor
- Competes with the substrate for enzyme
- Binds to the active site
- Inhibition is reversible
- ↑ substrate conc. = ↓ inhibitor conc.
2. Noncompetitive Inhibitor
- Binds to the allosteric site = shape of the active
site is changed
- ↑ substrate conc. does not reverse inhibi on
3. Uncompetitive Inhibitor
- Binds to the E-S Complex
- ↑ substrate conc. = ↑ inhibi on process
- E-S-I complex does not yield product