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Lecture 7

Topic 8
Enzymes

Topic 8
Enzymes

1) Introduction

2) Enzyme Kinetics

3) Effect of temperature on enzymes

4) Effect of pH on enzymes

5) Immobilized enzyme system

Main reference : Shuler and Kargi, Bioprocess Engineering

Additional reading : Homaei et al., Enzyme immobilization: an update, J
Chem Biol (2013) 6:185–205

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Introduction

Please refer to Table 3.1 & 3.2 for classification of enzyme

Please refer to Topic 5 (lecture 4)

Introduction – How enzymes work

Please refer to Topic 5 (lecture 4)

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Introduction – How enzymes work

Please refer to Topic 5 (lecture 4)

Enzyme Kinetics

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Enzyme Kinetics

Enzyme Kinetics : Michaelis-Menten kinetics

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Enzyme Kinetics : How to determine Km and Vmax

Enzyme Kinetics : How to determine Km and Vmax

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Enzyme Kinetics : How to determine Km and Vmax

Enzyme Kinetics : Inhibited enzymes kinetics

Please refer to Topic 5 (lecture 4) for type of inhibition

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Enzyme Kinetics : Inhibited enzymes kinetics

Equation 3.28 -3.38

Enzyme Kinetics : Inhibited enzymes kinetics

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Effect of temperature on enzymes

Effect of pH on enzymes

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Immobilized enzyme system

Immobilization
The containment of enzyme solution within a confined space for
the purpose of retaining and re-using enzyme in processing
equipment. There are many advantages that accompany
immobilized enzymes and many methods for immobilization.
Advantages
1. Reduce costs of operation compared to free enzyme systems
where additional separation and purification steps are needed.
2. Some immobilization methods can increase enzyme activity.
3. A model system to study enzyme action in membrane-bound
enzymes that occur in the cell.
Disadvantages
1. Many immobilized enzymes exhibit lower activity compared to
free enzymes.
2. More expensive to prepare than free enzymes.
3. Mass transfer limitations due to immobilization methods.

Immobilized enzyme system

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Immobilized enzyme system

Examples of immobilization technique

Source: Homaei et al., J Chem Biol (2013) 6:185–205

Immobilized enzyme system

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Immobilized enzyme system

Immobilized enzyme system

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Immobilized enzyme system

Additional reading

Immobilized enzyme system

Source: Homaei et al., J Chem Biol (2013) 6:185–205

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Production of enzyme

Utilization of enzymes

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Utilization of enzymes

Source: Homaei et al., J Chem Biol (2013) 6:185–205

Utilization of enzymes

Source: Homaei et al., J Chem Biol (2013) 6:185–205

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Utilization of enzymes

Source: Homaei et al., J Chem Biol (2013) 6:185–205

Exercise 1

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Exercise 1

[S](u V(nmol/
M) min) 1/[v] 1/[S]
0.2 1.43 0.70 5.00
0.26 1.67 0.60 3.85
0.33 2.08 0.48 3.03
1 3.33 0.30 1.00

Exercise 2

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Exercise 2
V1 V2 V3
I = 0.25 I = 50
[S] I = 0 nM nM nM 1/[S] 1/V1 1/V2 1/V3
0.4 0.22 0.21 0.2 2.50 4.55 4.76 5.00
0.67 0.29 0.26 0.24 1.49 3.45 3.85 4.17
1 0.32 0.3 0.28 1.00 3.13 3.33 3.57
2 0.4 0.36 0.32 0.50 2.50 2.78 3.13

Announcement

1) Tutorial for Topic 8 to be uploaded in smart2ums.

2) Tutorial (28th April) starts at 8.30 am.

3) Group project to be uploaded in smart2ums.

4) Midterm exam : 4th May 2017, 10 am (to be

confirmed), Topic 6 - 8

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