Enzymes

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Enzymes
• Biological catalyst
• Most of them are proteins
• They are neither consumed nor permanently altered after the
reaction
• Highly selective
• Enzymes are highly effective catalysts, commonly enhancing reaction
rates by a factor of 105 to 1017 .

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• Some enzymes require no chemical groups for activity other than their
amino acid residues. Others require an additional chemical component
called a cofactor – inorganic ions, or coenzyme – a complex organic or
metalloorganic molecule

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• Prosthetic group- a coenzyme or metal ion that is very tightly or even
covalently bound to the enzyme protein
• Holoenzyme- a complete, catalytically active enzyme together with its
bound coenzyme and/or metal ions
• Apoenzyme or Apoprotein- an enzyme without its characteristic
prosthetic group

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Enzyme Classification
1. Oxidoreductases – catalyze oxidation and reduction
a. Oxidases
b. Reductases
c. Dehydrogenases
2. Transferases – transfers a group from a donor to an acceptor
molecules
a. Transaminases
b. Kinases

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3. Hydrolases – hydrolysis reactions
a. Lipases
b. Proteases
c. Nucleases
d. Carbohydrases
e. Phosphatases
4. Lyases – addition of groups to double bonds, or formation of double
bonds by removal of groups
a. Dehydratases
b. Decarboxylases
c. Deaminases
d. Hydratases
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5. Isomerases – transfer of groups within molecules to yield isomeric
forms
a. Racemase
b. Mutases
6. Ligases – Formation of C-C, C-S, C-O, and C-N bonds by condensation
reactions coupled to ATP cleavage
a. Synthetases
b. Carboxylases

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• Enzyme Commission number (E.C. number)
Ex.

Enzyme - ATP:glucose phosphotransferase

E.C. number – 2.7.1.1.
Class 2 – Transferase
7 – Transfer of phosphoryl group
1 – Alcohol is phosphoryl acceptor
1 – D-glucose as the phosphoryl group acceptor
Trivial name – hexokinase

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• Active site – a crevice-like location in the enzyme involved in substrate
interaction
• Substrate – the molecule that is bound in the active site and acted
upon by the enzyme

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• Spontaneous Reaction • Catalysts enhance reaction rates by
lowering activation energies.

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• Role of binding energy in catalysis.

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Enzymatic Models
1. Lock and Key Model – based on the shape and fit between
substrate and active site; coupled with weak binding forces

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2. Induced-fit Model – considers enzymes as not rigid and static, and
allows for enzyme flexibility in their shapes

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Enzyme Activity
• Enzyme activity is a measure of the rate at which an enzyme converts
substrate to products in a biochemical reaction
• Factors which affect enzyme activity
1. Temperature
2. pH
3. Substrate concentration
4. Enzyme concentration
• At constant enzyme concentration, increasing the substrate
concentration will produce a saturation curve

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Enzyme Kinetics

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Michaelis-Menten Equation
Vmax [S]
Vi= Km+[S]

• Vi - initial rate (or initial velocity)

• Vmax – maximum velocity
• Km – substrate concentration at which
Vi equals half of Vmax

[S] = Km, Vi = ½ Vmax

Low Km; enzyme has high affinity for its
substrate
High Km; enzyme has low affinity for its
substrate
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Lineweaver-Burk equation
• Reciprocal of Michaelis-
Menten Equation. Linear
equation which allows for
the determination of the
Vmax and Km. May also
be used to determine the
specific type of inhibition

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1 Km 1
= +
Vi Vmax [S] Vmax

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Enzyme Inhibition
• Enzyme inhibitor- a substance that slows or stops the normal catalytic function of an
enzyme by binding to it.
• Types of Inhibition:
1. Irreversible – forms strong covalent bond to an amino acid in the active site and
deactivates the enzyme
2. Reversible Inhibition
o Competitive – resembles the enzyme’s substrate and competes with it for occupancy of
the active site

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o Uncompetitive – binds only to an enzyme-substrate complex

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oNoncompetitive – binds to a site other than the active site
– interact with both E and ES

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Inhibitor type Effect on Vmax Effect on Km
Competitive no effect increase
Uncompetitive decrease decrease
Noncompetitive decrease no effect

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Example
Determine the values for Vmax and Km for this enzymatic system with
and without the inhibitor. From those values, classify the type of
inhibitor

[S], mM No inhibitor With inhibitor

V, mmol/min V, mmol/min
3.0 4.58 3.66
5.0 6.40 5.12
7.0 7.72 6.18
9.0 8.72 6.98
11.0 9.50 7.60

Vmax = __________ Km = ___________Inhibitor Type = ___________

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