Biochemistry LEC

Enzymes

OUTLINE

I. What are enzymes?

II. Characteristics
III. Classification
IV. Cofactor
V. How does enzyme work?
VI. Enzyme kinetics
VII. Factors that affect enzymatic reactions
VIII. Zymogen
IX. Clinical Uses

I. What are Enzymes?

- Enzymes are proteins that serve as biological catalysts (meaning, if they are not present in the metabolic
process, then the process itself will not occur) for reactions in all living organisms
- Increase the rate of reactions, but are not permanently changed in the process
- Crucial to the biological reactions that occur in the body
- In humans, enzymes must catalyze reactions under very specific physiological conditions
o pH = 7.4
o Temperature = 37C
- Enzymes work well in neutral pH and temperatures that are the same as human temperatures
- Without them, metabolic process will not occur. In glycolytic process, there are several enzymes that needs to
be present in order for us to convert your glucose into your pyruvate.

II. Characteristics of Enzymes

- Enzymes greatly enhance reaction rates
- An enzyme-catalyzed reaction can be 106 to 1012 times faster than a similar uncatalyzed reaction
- Enzymes are very specific – but it varies (meaning, enzymes work at a specific metabolic process but some
enzymes also work for example: in another metabolic process. But for example, this particular set of enzymes
work for the glycolytic pathway or gluconeogenesis, then dun lang sila magwowork.)
III. Classification of Enzymes

Classification Description Enzymes

Oxidoreductases Catalyze an oxidation-reduction reaction Lactate Dehydrogenase (LD)
between 2 substrates Glucose-6-phosphate dehydrogenase
(G6PD)
Transferases Catalyze the transfer of a group other than Aspartate aminotransferase
hydrogen from one substrate to another (AST/GOT)
Alanine aminotransferase (ALT/GPT)
Creatinine kinase (CK)
y – glutamyltransferase (GGT)
Hydrolases Catalyze hydrolysis of various bonds Alkaline phosphatase (ALP)
Acid phosphatase (ACP)
Amylase (AMS)
Lyases Catalyzes removal of groups when substrates Aldolase (ALD)
 without hydrolysis, the product contains double
bonds
Isomerases Catalyzes the interconversion of geometric, Phosphohexose Isomerase
optical, or positional isomers
Ligases Catalyzes the joining of 2 substrate molecules Glutathione synthetase (GSH-S)
*kinase – use for the transfer of phosphate groups

*isomerases do not conform or make another group, rather, they just change the structure of your group or compound
that you are supposed to change. In glycolytic pathway that serves as an isomerase is phosphoglucose isomerase or
phosphohexose isomerase (yung nagconvert ng glucose-6-phosphate to fructose-6-phosphate) (they do not change the
compound itself, rather, they just change the structure, positions)

IV. Cofactor
- Metal ion or a nonprotein organic molecule needed for an enzyme-catalyzed reaction to occur
- Coenzyme – an organic compound that serves as an enzyme cofactor

V. How does enzymes work?

- Enzymes contain a region called an active site that binds the substrate, forming an enzyme-substrate complex
(in order for us to have a reaction, you must have an enzyme and a substrate. Once they combine together, they
will now form your enzyme substrate complex)
- Two-Models on how enzymes work
o To explain the specificity of a substrate for an active site
 Lock-and-Key Model
 Induced-Fit Model

Lock-and Key Model

- The shape of the active site is rigid

- 3D geometry of the substrate must exactly match the shape of the active site for catalysis to occur
- Explains high specificity in enzyme reactions

Enzyme-substrate complex
Induced-Fit Model

- Shape of the active side is flexible

- When substrate and enzyme interact, the shape of the active site can adjust to fit the shape of the substrate

VI. Enzyme Kinetics

- Chemical reaction may occur spontaneously if the free energy or available kinetic energy is higher for the
substrate than product
- Enzymes catalyze physiologic reactions by lowering the activation energy level that the substrate must reach for
the reaction to occur
- An enzyme combines only with 1 substrate and catalyzes only 1 reaction – ABSOLUTE SPECIFICITY
- Group specificity – enzymes combines with all substrates in a chemical group
- Bond specificity – enzymes combine with specific chemical bonds

VII. Factors that affect enzymatic reactions

- Zero-order reaction – reaction rate depends only on enzyme concentration (The more enzymes you have, then
the more reactions occurring)
- First-order reaction – reaction rate is directly proportional to substrate concentration (you will need a sufficient
amount, or balance out the first order reaction, so you have to produce an enzyme)

Factors that affect enzymatic reactions:

Enzyme Concentration

- The higher the concentration, the faster the reaction

- More enzyme is present to bind with the substrate

Substrate Concentration

- With the amount of enzyme exceeding the amount of substrate, the reaction rate steadily increases as more
substrate is added
- When substrate concentration reaches a maximal value, higher concentration of substrate no longer result in
increased rate of reaction.

pH or Hydrogen Ion Concentration

- Most physiologic reactions occur in pH 7-8

 - Extreme pH levels may denature enzyme or influence its ionic state
o Changes in structure
o Change in charge of amino acid residue charge in the active site

Temperature

- Active at 25C, 30C, or 37C

- 37OC – optimum temperature for enzymatic activity
- Increased temperature increases rate of chemical reaction
- 60-65OC may result to denaturation of enzymes
- Temperature coefficient – for every 10OC increase in temperature, there is a two-fold increase in enzyme
activity (ex. 20OC to 30OC – there would be a 40x reaction increase in enzyme activity)

Inhibitors

- Enzymatic reactions may not progress if an inhibitor interferes with the reaction (inhibitors are also polypeptide
or proteins that may affect our enzyme-substrate complex)
- Competitive inhibitor
o Physically binds to the active site of an enzyme
o Both substrate and inhibitor compete for the same active site
o Once the inhibitor binds to the active site first, then the reaction would not occur.

- Non-competitive inhibitor
o do not compete with the substrate for the active site
o Binds to the allosteric site (Wherein the substrate can still bind to the active site, however this causes
the reaction to be slower)
o Slower rate of reaction
 - Uncompetitive inhibitor
o Binds to the enzyme-substrate complex
o Increasing substrate concentration results in increasing the inhibition (when enzyme-substrate complex
is complete, that is the time the inhibitor binds to it)

VIII. Zymogen
- Enzymes in their inactive forms
- aka Proenzyme
- Contains additional amino acids in the chain and cleaved when activated

IX. Clinical Uses

Clinically…

- Enzymes are found in all body tissues

- They frequently appear in the serum following cellular injury and degraded cells
- Plasma/Serum enzyme levels are often useful in diagnosis of particular diseases

Enzymes in Medicine: DIAGNOSIS

- Measuring enzyme levels in the blood and understanding the key role of enzymes in biological reaction have
aided greatly in both diagnosing and treating disease
- The most commonly tested enzymes in the laboratory are as follows:

ENZYME CLINICAL SIGNIFICANCE

Acid Phosphatase (ACP) Prostatic carcinoma
Alanine aminotransferase (ALT) Hepatic Disorder
Aldolase (ALD) Skeletal muscle disorder
Alkaline phosphatase (ALP) Hepatic disorder
Bone disorder
Amylase (AMS) Acute pancreatitis (Fast marker)
Angiotensin-converting enzyme (ACE) Blood pressure regulation
Aspartate aminotransferase (AST) Myocardial infarction
Hepatic disorder
Creatinine kinase (CK) Myocardial infarction
Glucose-6-phosphate-dehydrogenase (G6PD) Drug-induced hemolytic anemia
y-glutamyltransferase (GGT) Hepatic disorder (associated w/ Alcoholic liver disease)
 Lactate dehydrogenase (LD) Myocardial infarction
Carcinoma
Lipase (LPS) Acute pancreatitis (specific marker)
Pseudocholinesterase (PChE) Organophosphate poisoning (Usually farmers poisoned
w/ insecticide)

Enzymes in Medicine: DRUGS

 ACE Inhibitors
o Group of drugs used to treat individuals with high blood pressure
o Narrows the blood vessels, thus increasing blood pressing
o Formed from zymogen angiotensin by action of ACE (angiotensin-converting enzyme)
 HIV protease inhibitors
o Inhibit the action of the HIV protease enzyme (enzyme needed by HIV to make copies) (within immune
cells, they use enzymes to make copies of themselves) (they inhibit the enzyme na nagpapaduplicate kay
HIV)
o Resulting in the… Decrease the virus population

*both are drugs that are to be intake

*paracetamol is an inhibitor that inhibits the pain, rather than to lower your body temp or fever. Kaya nag increase ng
body temperature because our cells increase heat in order to fight back kung ano man yung pathogen or nagiinfect
satin. As long as mataas ang temp, may pathogen sa body natin. Yun ang mechanism ng cells natin kaya tumataas ang
body temp natin. Paracetamol most likely used to inhibit the pain. They block pain receptors, and then we will not
feel the pain.