Saint Tonis College, Inc.

Formerly Kalinga Christian Learning Center

Purok 4, Bulanao, Tabuk City, Kalinga 3800
sherryannhmanganip@gmail.com

Course Subject : Biochemistry

Course Topic : Enzymes

Learning Objectives:

At the end of the topic, the students will be able to:

1. Define Enzyme.
2. Know the general structural classes of Enzymes.
3. Discuss the different model of enzyme activity.
4. Enumerate the discuss the different classification of Enzymes.
Learning Discussion
Enzymes
Enzymes- Greek word en- “in”, and zyme-yeast
Characteristics of Enzymes:
 Biocatalysts, mostly protein in nature, that enable the necessary metabolic reaction to occur at body
temperature, speeding up reaction rates
Ribozyme- some biocatalysts are made of RNA
 Acts by decreasing activation energy required for the biochemical reaction to push through
 Affects only the rate of the reaction
 They are not used up or consumed in the reaction.
 Most chemical reaction in the body occur at pH 7 and temperature of 37°C.
2 General Structural Classes of Enzyme
1. Simple Enzyme- composed only of protein (amino acid chains)
2. Conjugated Enzyme- enzyme that has nonprotein part in addition to protein part
Enzymology – Definition of Terms
 Haloenzyme- whole enzyme molecule
 Apoenzyme - protein portion of the enzyme
 Cofactor - non-protein portion of the enzyme; can be organic or inorganic
 Coenzyme - organic cofactor
Examples: NAD/NADH, Pyridoxal phosphate
 Activators- inorganic cofactors; can be metallic or non-metallic
 Metallic Calcium, Magnesium, Zinc, Iron, Manganese, Copper
 Non-metallic Chloride
 Zymogen/Proenzyme - inactive form of enzyme
e.g. pepsinogen, plasminogen
 Isoenzyme- enzymes that catalyse the same reaction but differ in terms of physical or chemical characteristics,
tissue distribution as well as electrophoretic mobility
e.g. lactate dehydrogenase- LD1, LD2, LD3, LD4, LD5
creatine kinase- CKMM, CKMB, CKBB
acid phosphatase- prostatic and erythrocytic ACP
 Macroenzyme- are high-molecular-mass forms of the serum enzyme
 Substrate- precursor of a product in an enzymatic reaction
-binds to the active site of the enzyme

Models of Enzyme Action

Enzyme Active Site - relatively small part of an enzyme’s structure that is actually involve in catalysis.
 Usually a “crevicelike” location in the enzyme
Enzyme-Substrate Complex - the intermediate reaction species that is formed when a substrate binds to the
active site of an enzyme.
1. Lock-and-Key Model- the active site in the enzyme has fixed,rigid and geometrical conformation.
o Only a substrate whose shape and chemical nature are complementary to those of the active
site can interact with the enzyme.
 2. Induced-Fit Model - allows for small changes in the shape or geometry of the active site of an enzyme to
accommodate a substrate.
o The enzyme active site, although not exactly complementary in shape of that of a substrate, is
flexible enough that it can adapt to the shape of the substrate

Enzyme Specificity- is the extent to which an enzyme’s activity is restricted to specific substrate, a specific group of
substrate, a specific type of chemical bond, or a specific type of chemical reaction.

 The degree of enzyme specificity is determined by the active site.

 Absolute Specificity - the enzyme will catalyse only one reaction
 Group Specificity - the enzyme will act only on molecules that have a specific functional group.
 Linkage Specificity - the enzyme will act on a particular type of chemical bond, irrespective of the rest of
the molecular structure
 Stereochemical Specificity - the enzyme will act on a particular stereoisomer.

Factors that Affect Enzymatic Reactions

 pH- excessively alkaline o acidic pH can affect the enzymes and can denature enzymes.
-some enzymes however, perform well in alkaline (e.g. ALP) and acidic (e.g. ACP and pepsin) environment.

 Temperature - reaction of most chemical reactions increase with temperature and approximately double for
each 10° rise
-most enzyme become denatured and insoluble within minutes of being subjected to temperatures
of 60° and above
 Enzyme Concentration - velocity of reaction is directly proportional to enzyme concentration in the presence of
excess substrate
 Substrate concentration
 Machaelis-Menten hypothesis
 The higher the substrate concentration, the more substrate bound to enzyme and the greater
the rate/ velocity of the reaction.
 When all enzyme is bound to substrate, there will be no further increase in velocity
 When the substrate is present in an adequate amount (all enzymes are bound and saturated),
the rate of the reaction depends only in the enzyme concentrate.
 First Order Reaction
 Substrate readily binds to free enzyme at a low-substrate concentration
 Rate depends on substrate concentrate
 Zero Order Reaction
 High substrate concentration saturate all available enzymes; fewer binding site are available for
attachment
 Reaction velocity reaches maximum ; rate falls to zero
 Rate depends only on enzyme concentration
 Inhibitors- substances that inhibit enzymatic reaction
 Competitive Inhibition - inhibitors binds the active site of enzyme.
 Substrate and inhibitors compete for the same binding site
 Can be reversed by the addition of substrate molecules.
  Noncompetitive Inhibition - inhibitor binds enzyme at a place other than the active site of enzyme.
 Alter the configuration of the enzyme in such a way as to reduce or abolish excess of the
substrate to the active site, causing a decrease in enzyme activity.
 May be reversible (if binding is reversible or may be separated) or irreversible (if inhibitor
destroys a part of enzyme)

 Uncompetitive Inhibition - inhibition binds the ES complex

 The resulting enzyme-substrate –inhibitor complex do not yield product.

 Ionic Strength or Electrolyte Environment
-if ionic strength is too high, enzyme activity drops
-if protein-free solution, enzymes lose activity rapidly.
Importance of Enzyme Detection:
 To detect tissue injury or damage
 To identify abnormalities or deficiency of enzymes
Measuring Enzymatic Activity
o Enzymes are measured based on their activity rather than the mass or molar concentration.
o Enzymatic activity can be measured as a rate of product produced or as a rate of substrate consumed
per unit of time.
o Units used in expressing enzymatic activity:
 Enzyme activity can be expressed either in IU or KU
 International Unit- micromole of substrate per minute:
 Katal Unit - mole of substrate per second
Principles of Enzyme Data Interpretation
1. There is no truly “organ-specific” enzyme- since all enzymes are found in more than one tissue.
2. Serial measurement provide most useful data; a single measurement can be misleading
3. Negative (normal) results are useful especially when ruling out presence of tissue damage
4. Enzyme data must be integrated with other information.
Classification of Enzymes
1. Oxidoreductases- oxidize one substrate and reduce the other.
2. Transferases- transfer a group (other than hydrogen)from substrate to another
3. Hydrolases- catalyse hydrolysis or splitting up of a bond in a substrate, resulting in the formation of two or more
molecules
4. Lyases- catalyse removal of group from substrate without hydrolysis; the product contains double bonds
5. Isomerases- catalyse the interconversion of geometric, optical or positional isomers
6. Ligases- catalyse the joining of two substrate molecules, coupled with breaking of the pyrophosphate bond in
adenosine triphosphate(ATP) or a similar compound.
Oxidoreductase Transferase Hydrolase Lyases Isomerase Ligases
(dehydrogenase) (kinase/transaminase) (decarboxylase)
*Lactate *creatine kinase *acid &alkaline *aldolase *glucose Glutathione
dehydrogenase *aspartate phosphatase *pyruvate phosphate synthetase
*Glucose-6- aminotransferase(AST) *cholinesterase decarboxylase isomerase
phosphate *alanine *lipase *glutamate *ribose
dehydrogenase aminotransferase(ALT) *trypsin decarboxylase phosphate
*Malate *gamma glutamyl *pepsin *tryptophan isomerase
dehydrogenase transferase(GGT) *leucine decarboxylase
*Isocitrate *hexokinase aminopeptidase
dehydrogenase *pyruvate kinase *amylase
*Cytochrome *glucosidase
oxidase *galactosidase
*Glutamate *elastase
dehydrogenase
*Beta-
hydroxybutyric
dehydrogenase

Post-test:
1. What will happen to enzymes when they are at high temperature?

2. What will happen to enzymes when they are at low temperature?

3. What is the name of the molecule or substance that the enzyme reacts with?

4. True or False. Change in pH alters the active site of an enzyme.

5. What is the nature of enzyme?

a. vitamin

b. lipids

d. carbohydrates

d. protein

6. Enzymes change the ____ of the chemical reaction.

a. type

b. rate

c. reactants

d. products

7. In an enzymatic reaction, the amount of ______ determines the amount of product produced.

a. catalyst

b. reactants

c. oxygen

d. enzyme

8. When all enzymes are bound to substrate, there will be no further increase in velocity is called?

A. maximum velocity

B. maximum tolerance

C. increased velocity

D. increased tolerance

9. What is the 'active site' of an enzyme, and how is it critical in how an enzyme functions?

10. Enumerate and discuss the enzyme inhibitors.