Lecture 4: Enzyme Technology

26 September 2019
Dr Lim Mee Wei (limmeewei@segi.edu.my)
Faculty of Engineering and Built Environment
SEGi University, Kota Damansara Campus
 Last updated: 04 Dec 2013

Enzyme Inhibition
• Certain compounds may bind to enzymes and reduce
their activity  Enzyme inhibitors
• Enzyme inhibitors may be reversible or irreversible.
▫ Irreversible compounds (heavy metals: lead, cadium, mercury,
etc):
 Form stable complex with enzyme and reduce enzyme activity.
 May be reversed using chelating agents such as EDTA and citrate
▫ Reversible compounds
 May dissociate easily after binding

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Enzyme Inhibition

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Enzyme Inhibition

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Enzyme Inhibition (Competitive)

Competitive inhibitors: Compete with substrate for active
site of the enzyme. Usually are substrate analogs.

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Enzyme Inhibition (Non Competitive)

Non Competitive inhibitors: Inhibitors bind on sites other
than active site, and reduce enzyme affinity to the
substrate. Not substrate analogs.

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Enzyme Inhibition (Uncompetitive)

Non Competitive inhibitors: Bind to ES complex only, and
no affinity for enzyme itself

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Enzyme Inhibition (Substrate Inhibition)

Substrate inhibitors: Occurs when there are high substrate
concentrations

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Worked Example 1
What type of inhibition reaction is the below? Substantiate the answer. Then, determine
the Michaelis-Menten equation, K’m, and the value of inhibitor constant, Ki.

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Worked Example 1
Plot a double reciprocal plot or Lineweaver-Burk plot of 1/v versus
1/[S] for inhibitor concentration I=0. I=0.0012 M, I=0.0044M and
I=0.0060.

Trend of the graph indicates that inhibition is noncompetitive.

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Problem!
• Enzymes are proteins. They are soluble in water. Thus, it
is hard to reuse them once the reaction is over. What is
your suggestion to overcome this problem?

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Immobilized Enzyme Technology

• Defined as restriction of enzyme mobility in a fixed
space.
• Advantages:
• enzyme reutilization

• elimination of enzyme recovery and purification

process
• provides better environment for enzyme activity

• Improves product purity

• Reduces effluent handling problems

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Immobilized Enzyme Systems

• Matrix entrapment
(a) (b)

Binding
•
(c) (d)

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Class Activity 1
• What are the advantages
and disadvantages of the
following immobilization
techniques?
▫ Entrapment
 Matrix entrapment
 Membrane entrapment
▫ Surface immobilization
 Adsorption
 Covalent bonding

Get into groups and discuss.

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Application of Immobilization technique in

Production of Biodiesel

enzyme

separation
Reactor

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Compulsory page

References:
Shuler, M. L. and Kargi, F. 2002, Bioprocess Engineering –
Basic Concepts, 2nd edn, Prentice Hall.

Katoh, S. and Yashida, F. 2009, Biochemical Engineering:

A Textbook for Engineers, Chemists and Biologists, Wiley-
VCH Verlag GmbH & Co. KGaA, Weinheim.

Rao, D. G. 2010, Introduction to Biochemical Engineering,

2nd end, McGraw Hill.

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