Student copy

CENTRO ESCOLAR UNIVERSITY

Manila Campus
School of Medical Technology
2nd Semester, 2024-2025

CLINICAL CHEMISTRY II

ENZYMES
• Enzymes are specific biologic proteins ____________________________________ without
altering the equilibrium point of the reaction or being consumed or changed in composition
• The catalyzed reactions are frequently specific and essential to physiologic functions such as:
o Hydration of CO2
o Nerve conduction
o Muscle contraction
o Nutrient degradation
o Energy use
• Found in all body tissues, enzymes frequently appear in the serum following cellular injury or,
sometimes, in smaller amounts from degraded cells.
• Certain enzymes, such as those that facilitate coagulation, are specific to plasma and, therefore, are
present in significant concentrations in plasma
• Plasma or serum enzyme levels are often useful in the diagnosis of particular diseases or
physiologic abnormalities.
• As a protein, each enzyme contains a specific amino acid sequence (primary structure), with the
resultant polypeptide chains twisting (secondary structure), which then folds (tertiary structure)
and results in structural cavities

PROPERTIES OF AN ENZYME
Each enzyme contains:

1. __________________________ is a water-free cavity, where the substance on which the

enzyme acts (the substrate) interacts with particular charged amino acid residues

2. __________________________ = is a cavity other than the active site; may bind regulator
molecules and, thereby, be significant to the basic enzyme structure.

_____________________________ – is a substance that an enzyme acts upon to create a final product

ENZYME THEORY
1. ______________________________________
o It is based on the premise that the shape of the key (substrate) must fit into the enzyme

2. --------------------------------------------------------
o It is based on the substrate binding to the active site of the enzyme

ENZYME NOMENCLATURE
• To standardize enzyme nomenclature, the Enzyme Commission (EC) adapted a classification
system in 1961, and revised the standards in 1972 and 1978.
• Enzymes are classified according to their biochemical functions, indicating substrate and class of
reaction catalyzed, and are designated by individual identification numbers
• The first digit, places the enzyme in its classification (sic classifications)
• The second and third digits, represents the subclass to which the enzyme is assigned
• The final and fourth number/s, is a serial number that is specific to each enzyme.

Classes of Enzymes
There are six classes of enzymes, describing the type of reaction involved:

1. Oxidoreductases
o Catalyze an _________________________________ between two substrates
2. Transferases
o Catalyze the _________________________________ from one substrate to another
3. Hydrolases
o Catalyze ____________________________________ by the addition of water (hydrolytic
reactions)
4. Lyases
o Catalyze ____________________________________________________. The product
contains double bonds
5. Isomerases
o Catalyze the ______________________________________ of the substrate compound
6. Ligases
o Catalyze the _____________________________________, coupled with breaking of the
pyrophosphate bonding in adenosine triphosphate (ATP) or similar compound

Factors affecting enzymatic reactions:

❖ Enzymatic Concentration
o The higher the enzyme concentration, the _____________________ is the reaction,
because more enzyme is present to bind with the substrate.

❖ Substrate Concentration
o With the amount of enzyme exceeding the amount of substrate, the reaction rate steadily
_____________________ as more substrate is added.
o However, when the substrate concentration reaches a maximal value, higher concentration
of substrate no longer results in increased rate of reaction.

❖ Cofactors – a nonprotein entities that must bind to particular enzymes before a reaction occurs.
 a. Coenzymes – is an organic compound (second substrates)
▪ Increasing its concentration will increase the velocity of an enzymatic reaction
▪ It is essential to achieve absolute enzymatic activity
• Example: NAD (Nicotinamide adenine dinucleotide) and NADP
(Nicotinamide adenine dinucleotide phosphate)
b. Activators – are inorganic ions which alters the spatial configuration of the enzyme for
proper substrate binding.
• Example: calcium, zinc, chloride, magnesium and potassium
c. Metalloenzymes – are inorganic ion attached to a molecule
• Example: catalase and cytochrome oxidase

❖ Inhibitors: different types of interferences with the function of an enzyme that may be reversible
or irreversible and lower the reaction rate of product formation

o Competitive inhibitors: __________________________________________ of the

enzyme and prevent formation of product, but have a higher Km than the preferred substrate
and can be overcome by addition of more substrate.
o Noncompetitive inhibitors__________________________________________ of the
enzyme than the substrate and so cannot be overcome by addition of more substrate, but
prevent formation of product despite the enzyme-substrate complex.
o Uncompetitive inhibitors: _________________________________________ and prevent
the formation of product.

Inhibitors are important in the human body because they regulate enzyme activity, aiding in disease
treatment and enhancing defense mechanisms.

❖ Isoenzymes:
o These are enzymes (polypeptide chains) having the same catalytic reactions but slightly
different molecular structures.

❖ Temperature:
o Enzymes are active at 25˚C, 30˚C, or _______________ (optimum temperature for
enzymatic activity).
o Increasing temp. usually increases the rate of a chemical reaction by increasing the movement
of the molecule.
o The rate of denaturation increases as the temperature increases, and is usually significant at
40˚C to 50˚C.
o 60 - 65˚C may result to inactivation of enzymes.
o ________________________________ – for every 10˚C increase in temperature, there will be
a two-fold increase in enzyme activity.

❖ pH or Hydrogen Ion Concentration:

 o Most physiologic reactions occur in the pH of _________________
o Extreme pH level may denature an enzyme or influence its iconic state resulting in structural
change or change in the charge of amino acid residue in the active site

❖ Storage:
o Low temperature (refrigeration/freezing) render enzymes reversibly inactive
o Repeated freezing and thawing tend to denature proteins and should be avoided
o _______________ = ideal temperature for preservation of enzymes (loner period of time)
o _______________ = ideal storage temp for substrate and coenzymes
o 22˚C or room temperature = ideal for storage of LDH (LD4 and LD5

❖ Hemolysis – Mostly increases enzyme concentration

❖ Latescence or milky specimen = decreases enzyme concentration

Enzyme Activity
Enzymes are measured in terms of:

1. Change in the substrate concentration

2. Change in the product concentration
3. Change in coenzyme concentration

Causes of Elevated Plasma Enzyme Levels

1. Impaired removal of enzyme from plasma
2. Increased permeability of cell membrane
3. Increased in the number of cells or the production of cells
4. Increased in the normal cell turnover
5. Decreased clearance of enzymes from the circulation
6. Tissue necrosis and degeneration – death of enzyme-containing cells

Prepared by:

Prof. Ryann H. Valenzuela, RMT, MSMT