BIOCHEMISTRY

BIOCHEMISTRY 2:

ENZYMES
MFAC1501 Foundations 2015
Dr Anne M Galea
School of Biotechnology and Biomolecular Sciences
Room 103B, Biological Sciences Building
Ph: (02) 9385 8156, E-mail: a.galea@unsw.edu.au
http://www.azur-diving.com/wp-content/uploads/enlarge/6.jpg
MRE/AMG 2015

Lecture Overview
What are enzymes and what is catalysis?
Formation of the ES complex
Catalysis at the active site
Factors affecting the rate of enzymecatalysed reactions

Enzyme inhibition
Therapeutics based on enzyme inhibition
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ENZYMES
LEARNING OBJECTIVES:
Describe the role of enzymes as biological catalysts and
explain the fundamental principles of enzyme action.

Explain how enzymes speed up the rate of a reaction without

affecting the equilibrium of the reaction.

Describe how an enzyme lowers the activation energy for a

reaction to occur.

List and describe the chemical and physical factors that can
influence enzyme activity (including enzyme inhibitors).

Describe the way in which enzyme inhibition can be employed

in drug-based therapeutics (using examples).
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ENZYMES: Introduction
Enzymes are biological catalysts or catalytic proteins.
A catalyst is a chemical agent that changes the rate of a
reaction without being consumed by the reaction.

Enzymes are highly specific:

- with respect to the reactions they catalyse.

- with respect to the choice of reactants (substrates).
Enzymes have great catalytic power (can increase rates
of reaction by > x106).
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ENZYMES: Introduction
Enzymes speed up the attainment of equilibrium without
affecting the position of equilibrium.

Many enzymes require other components for activity

(e.g. cofactors or coenzymes).

In the absence of enzymes, chemical reactions within

cells would not occur rapidly enough to sustain life.

Representing an enzyme catalysed reaction:

Substrate(s)
S
Sucrose

enzyme
sucrase

Product(s)
P
Glucose + Fructose
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How Do Enzymes Speed Up Reactions?

Enzymes lower the energy barrier

(activation energy) of a thermodynamically
favourable reaction, thus increasing the rate
of that reaction. This is called catalysis.

Enzymes DO NOT affect the equilibrium or

free energy change (G) of the reaction and
cannot make a thermodynamically
unfavourable reaction favourable!

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Enzymes Accelerate Reactions by

Lowering Energy Barriers
Enzymes decrease the activation energy (EA) of a
reaction by providing an alternative reaction pathway to
the transition state

This reduced uphill

climb means that the
transition state is
reached more frequently
and the reaction
proceeds more rapidly
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How Do Enzymes Speed Up Reactions?

Campbell, Reece, Meyers Biology 8e 2009 Pearson Education Australia

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Formation of the ES Complex

E+S

ES the first step in enzyme catalysis

Induced Fit

Enzymes have active sites where catalysis takes place.

Part of the active site is the substrate binding site.
Substrate binding may result in conformational changes
(an induced fit model).
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Enzyme-substrate Complexes

Campbell, Reece, Meyers Biology 8e 2009 Pearson Education Australia

Note the conformational change that is induced as glucose

enters the active site of the enzyme, hexokinase, above.

Structures are determined by X-ray crystallography.
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Enzyme Catalysis
E+S

ES

E+P
S

A single enzyme typically

catalyses thousands of
reactions per second
E
ES

Release of
products

P
The enzyme is
not altered

EP
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Representing Enzymes: CPK Model

Lysozyme

Carbon grey
Nitrogen blue
Oxygen red
Sulphur yellow
All protein atoms are
shown as spheres with
radii approximately equal
to radii of atoms.
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Representing Enzymes: Ribbon Model

Triose Phosphate
Isomerase

Alpha helices shown as helices

Beta-sheets shown as arrows

Drawn by Jane
Richardson
IJM/AMG
2008
IJM/AMG
AMG
2009
2015

Factors Affecting Enzyme Activity

The physical and chemical environment of a cell
affects enzyme activity.

Some of the main factors affecting enzyme activity

include:
Enzyme concentration [E]
Substrate concentration [S]
Temperature
pH
Cofactor requirements
Inhibitors

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Initial rate of reaction

(v)

Enzyme Concentration

The rate of reaction is

directly proportional to the
enzyme concentration

[E]

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Substrate Concentration
A plot of the reaction
velocity (v0) as a
function of the substrate
concentration [S].

Michaelis - Menten
equation:
V0 = Vmax [S]
KM + [S]

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Temperature and pH

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Cofactor Requirements
Enzymes often require a non-protein

Coenzyme B12

component for activity.

Cofactors may be metal ions or more

complex organic molecules
(coenzymes).

Water soluble vitamins (B vitamins

and vitamin C) are either coenzymes
or converted into coenzymes.

This is the primary role of vitamins in

the human body and why they are
essential in the diet.

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Enzyme Inhibition
Enzymes can be inhibited by specific molecules
Competitive inhibitors mimic
the substrate, bind at the active
site and block substrate binding.

Non-competitive inhibitors bind

at a separate site, preventing
catalysis.

Inhibition is commonly
reversible (like substrate
binding, but may be irreversible
if a covalent bond is formed
between the inhibitor and an
amino acid side chain.
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Competitive Inhibition
E + S

ES

E + P

+
I

EI

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Non-competitive Inhibition
E + S

ES

E + P

+
S

I
E

ESI

Enzyme inhibition forms

the basis of action of many
pharmaceutical drugs

I
E

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Methotrexate Inhibits Dihydrofolate Reductase

Methotrexate is a close
structural analogue of
the substrate (folic acid)
and product and binds
tightly at the active site.

It is used in combination
therapy for cancers such
as leukaemia because
the enzyme is
particularly important in
rapidly dividing cells.

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Penicillin
Penicillin irreversibly inhibits an enzyme involved in
bacterial cell-wall synthesis

Inhibitor

Substrate

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Crixivan Inhibits the HIV Protease Enzyme

crixivan

HIV proteins are initially synthesised as part of a very large

polyprotein. This must be split into the different component
proteins by the HIV protease before they fold correctly and are
functional. Inhibiting the protease prevents HIV from being
infectious.
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READING
ENZYMES
Biochemistry, 7th Edition
Berg, Tymoczko & Stryer, 2012
Chapter 8 (pp.219-248)

Essential Biochemistry
Pratt & Cornely, 2004
Chapter 7 (pp.198-231)

Fundamentals of Biochemistry,
4th Edition
Voet, Voet & Pratt, 2013
Chapters 11 and 12

Biochemistry A Short
Course, 2nd Edition
Tymoczko, Berg & Stryer, 2013
Chapters 6, 7 and 8

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