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CC 2 Lab Ex 2

The document outlines Experiment 2 on Enzyme Kinetics, covering the general characteristics, classification, and basic concepts of enzymes. It discusses factors influencing enzymatic reactions such as substrate concentration, pH, temperature, cofactors, and inhibitors, along with the Michaelis-Menten curve and Lineweaver-Burk plot. The content is structured to provide a comprehensive understanding of enzyme behavior and kinetics in biochemical reactions.

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4 views

CC 2 Lab Ex 2

The document outlines Experiment 2 on Enzyme Kinetics, covering the general characteristics, classification, and basic concepts of enzymes. It discusses factors influencing enzymatic reactions such as substrate concentration, pH, temperature, cofactors, and inhibitors, along with the Michaelis-Menten curve and Lineweaver-Burk plot. The content is structured to provide a comprehensive understanding of enzyme behavior and kinetics in biochemical reactions.

Uploaded by

trishamacaranasmd
Copyright
© © All Rights Reserved
Available Formats
Download as PDF, TXT or read online on Scribd
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CLINICAL CHEMISTRY 2 (CC2)

8424 / LECTURE / BSMLS-3

EXPERIMENT 2: Enzyme Kinetics

Outiline
I. General Characteristics of Enzymes
II. Enzyme Classification and Nomenclature
A. Oxidoreductases
B. Transferases
C. Hydrolases
D. Lyases
E. Isomerases
F. Ligases
III. Basic Concepts of Enzyme Kinetics
IV. Factors that Influence Enzymatic Reactions
A. Substrate and Enzyme Concentration
B. pH
C. Temperature
D. Cofactors
E. Inhibitors
V. Other Concepts
A. Michaelis- Menten Curve
B. Lineweaver-Burk Plot

Experiment 2

Enzyme Kinetics

I. General Characteristics
➢ Active Site
- Where substrate and enzyme interacts
➢ Allosteric Site
- Site other than the active site
➢ Cofactors
- Activate or enhance the enzymatic reactions
■ Inorganic Cofactors
■ Organic Cofactor
➢ Proenzyme / Zymogen
- Inactive form; not capable in binding
➢ Holoenzyme
- Active form
➢ Isoform III. Basic Concepts
- Functionally related forms; not catalyzes same reaction ➢ Enzymes
➢ Isoenzyme ○ Specific biologic proteins
- Catalyzes same reaction but different in function ○ Catalyzes biochemical reactions
➢ Enzyme Kinetics
II. Classification and Nomenclature ○ Study of enzyme reactions and factors affecting it
○ Fisher’s Lock and Key Model
O 1. Oxidoreductases L 4. Lyases - Substrate must be specific to enzyme

T 2. Transferases I 5. Isomerase

H 3. Hydrolases L 6. Ligases

7 | TMM | S.Y. 24-25 | Prelims |


LABORATORY
■ Michaelis-Menten Equation
IV. Factors that Influence Enzymatic Reactions
1. Substrate and Enzyme Concentration

■ What if an enzyme can bind more than one substrate?


- Low Km = low substrate concentration; high binding capacity [only few
substrate is occupying the active sites]
- High Km = High substrate concentration; low binding capacity
➤ Lineweaver-Burk Plot
- Double reciprocal of the Michaelis-Menten
- Used for the determination of Vmax and Km
2. pH
- Optimum pH - Most favorable pH value
- Most enzymes react at pH 7.0-8.0
- Use of buffer solutions [maintains pH]
■ ACP - pH 5.0
■ ALP - pH 9.0-10.0
3. Temperature
- Optimum temperature: 37℃
- Directly proportional to rate of reaction
- An increase of 10 ℃ = doubles the rate of reaction
4. Cofactors
- Enzyme activity may NOT occur in the absence of cofactors
- In excess - inhibits the activity
- Presents in the minimal amounts
5. Inhibitors
- Competitive Inhibition
■ Inhibitor is competing to the same active sites
- Non-competitive Inhibition
■ Inhibitor binds to other sites than active site
- Uncompetitive Inhibition
■ Inhibitor binds to enzyme-substrate complex [change in Km and Vmax]

V.Others

➤ Michaelis - Menten Curve


➢ Describes the relationship between substrate concentration and rate of velocity
■ First Order Kinetics
- The substrate concentration is directly proportional to the reaction rate
- The higher the substrate concentration the higher the reaction rate
■ Zero Order Kinetics
- Plateu
- A chemical reaction where the rate of the reaction is independent of the
concentration of the reactants.
- This means that even if you increase or decrease the amount of the
starting materials, the reaction will proceed at a constant rate.

8 | TMM | S.Y. 24-25 | Prelims |


LABORATORY
INHIBITION

9 | TMM | S.Y. 24-25 | Prelims |

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