CHAPTER 1:

ENZYME KINETICS AND APPLICATIONS

(Part 1a : Kinetics of Enzyme Catalyzed Reactions)
ERT 317 Biochemical Engineering
Sem 1, 2016/2017

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 Role of Biochemical /Bioprocess
Engineer
exploit advances in biology to create new products
design biochemical processes & operate plants
develop energy resources
Develop new, environmentally friendly, and safer
processes to make the biochemical products that people
depend on.
Work in research and development laboratories,
creating polymeric materials with improved performance
and durability.
Work in manufacturing, making vaccines and antibiotics.
Invent new ways to keep our food and water supplies
safe.

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Bioprocess Engineers Task
Minimize production of unwanted
byproducts
Separate the good (product) from
the bad (byproducts)
Recover the unused reactants
Maximize profit, minimize energy
consumption
Minimize impact on the
environment

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 OUTLINE
Introduction
Enzyme Structure
Enzyme Function

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 Enzymes
There are many chemical compounds in the living cell.

How they are manufactured and combined at sufficient

reaction rates under relatively mild temperature and
pressure?

How does the cell select exactly which reactants will be

combined and which molecule will be decomposed?

Catalysis by ENZYME

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 Enzymes
Enzymes are biological catalysts that are protein molecules in
nature- react in mild condition

They are produced by living cells (animal, plant, and

microorganism) and are absolutely essential as catalysts in
biochemical reactions.

Almost every reaction in a cell requires the presence of a

specific enzyme related to its particular protein structure.

A major function of enzymes in a living system is to catalyze

the making and breaking of chemical bonds.

Therefore, like any other catalysts, they increase the rate of

reaction without themselves undergoing permanent chemical
changes.

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 Over 2000 enzymes have been identified

Often named by adding the - ase to the name of

substrate acted upon, or the reaction catalyzed
such as urease, alcohol dehydrogenase

The majority of cellular reactions are catalyzed by

enzymes

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 Some protein enzyme required a non-protein group for their activity.

Non protein group:

Cofactors: metal ions, Mg, Zn, Mn, Fe.
Coenzyme: complex organic molecule, NAD, FAD, CoA
Vitamins

Catalyze biochemical reactions

breaking, forming and rearranging bonds.

Catalytic function very specific and effective (Specific because of

conformational shape)
Dictated by the enzyme active site.
Some active sites allow for multiple substrates.

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 Enzymes are catalysts
Catalyst: chemical that changes the rate of a reaction without
being consumed
Recycled (used multiple times)

Enzymes reduce the activation energy of a reaction

Amount of energy that must be added to get a reaction to
proceed

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 Catalysts
A catalyst is unaltered during the course of a reaction and functions in
both the forward and reverse directions.

In a chemical reaction, a catalyst increases the rate at which the reaction

reaches equilibrium.

For a reaction to proceed from starting material to product, the chemical

transformations of bond-making and bond-breaking require a minimal
threshold amount of energy, termed activation energy.

Generally, a catalyst serves to lower the activation energy of a particular

reaction.

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 Enzyme Function

Enzymes lower the activation energy of reaction catalyzed

( They do this by binding to the substrate of the reaction, and

forming an enzyme-substrate (ES) complex)

Substrate binds to a specific site on the enzyme called the active

site

Multi-substrate reactions possible

Lock and key model

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 The activation
energy for the
decomposition of
hydrogen
peroxide varies
depending on the
type of catalysis.

Type of catalysis Activation

energy

Uncatalyzed 18
reaction at 20C kcal/mol
Enzymatically 7 kcal/mol
catalyzed
(catalase)
Enzyme lower the activation energy of the reaction by binding
Chemically 13
catalysed (by kcal/mol the substrate and forming an enzymes-substrate complex.
collodial
platinum)
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Comparison of activation energies in the uncatalyzed and
catalyzed decompositions of ozone.

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 Important Terms To Be Remember!

active site - a region of an enzyme comprised of different amino

acids where catalysis occurs or a small portion of the surface of
an enzyme which a specific chemical reaction is catalyzed

substrate - the molecule being utilized and/or modified by a

particular enzyme at its active site

co-factor - organic or inorganic molecules that are required by

some enzymes for activity. These include Mg2+, Fe2+, Zn2+ and
larger molecules termed co-enzymes like nicotinamide adenine
dinucleotide (NAD+), coenzyme A, and many vitamins.

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 Types of Enzymes

holoenzyme - a complete, catalytically active enzyme

including all co-factors OR an enzyme containing a non
protein group

apoenzyme - the protein portion of a holoenzyme minus the

co-factors OR the protein part of holoenzyme

(holoenzyme = apoenzyme+cofactor)

isozyme - (or iso-enzyme) an enzyme that performs the same

or similar function of another enzyme that occur in several
different molecular forms.
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 Nomenclature of enzyme
Originally enzymes were given non descriptive names such as:
rennin : curding of milk to start cheese-making processor
pepsin : hydrolyzes proteins at acidic pH
trypsin : hydrolyzes proteins at mild alkaline pH

The nomenclature was later improved by adding the suffix -ase to the name of the substrate with which the
enzyme functions, or to the reaction that is catalyzed, for example:

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Alcohol
dehydrogenase

Glucose isomerase

Glucose oxidase

Lactic acid
dehydrogenase

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 Enzyme reactions are different from
chemical reactions, as follows:
1. An enzyme catalyst is highly specific, and catalyzes only one or
a small number of chemical reactions. A great variety of enzymes
exist, which can catalyze a very wide range of reactions.

2. The rate of an enzyme-catalyzed reaction is usually much faster

than that of the same reaction when directed by nonbiological catalysts
at mild reaction condition.

3. A small amount of enzyme is required to produce a desired effect.

4. Enzymes are comparatively sensitive or unstable molecules

and require care in their use.

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 Enzymatic Reaction Principles
Biochemically, enzymes are highly specific for their substrates and
generally catalyze only one type of reaction at rates thousands and
millions times higher than non-enzymatic reactions.

Two main principles to remember about enzymes are :

a) they act as CATALYSTS (they are not consumed in a
reaction and are regenerated to their starting state)

a) they INCREASE the rate of a reaction towards equilibrium

(ratio of substrate to product), but they do not determine the
overall equilibrium of a reaction.

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 Reaction Rates
The rate of the reaction is effected by several factors
including the concentration of substrate, temperature and
pH.
For most standard physiological enzymatic reactions, pH
and temperature are in a defined environment (eg; pH 6.9-
7.4, 37oC).
This enzymatic rate relationship has been described
mathematically by combining the equilibrium constant, the
free energy change and first or second-order rate theory.
Keq = eGo/RT
The net result for enzymatic reactions is that the lower the
activation energy, the faster the reaction rate, and vice versa.

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 Specificity
Most synthetic catalyst are not specific
i.e., they will catalyze similar reactions
involving many different kinds of
reactants.

While enzymes are specific. They will

catalyze only one reaction involving only
certain substances.

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 Binding Energy
The interaction between enzyme and its
substrate is usually by weak forces.

In most cases, Van der Waals forces and

hydrogen bonding are responsible for
the formation of ES complexes.

The substrate binds to a specific site on

the enzyme known as the active site.

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 Classification of Enzyme
Enzymes fall into 6 classes based on function

1. Oxidoreductases: which are involved in oxidation, reduction, and

electron or proton transfer reactions

2. Transferases : transfer of functional group

3. Hydrolases : which cleave various covalent bonds by hydrolysis

4. Lyases : catalyse reactions forming or breaking double bonds

5. Isomerases : catalyse isomerisation reactions

6. Ligases : join substituents together covalently.

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 References

Shuler, M. L. and Kargi, F. (2014). Bioprocess Engineering: Basic

Concept. 2nd Ed. Upper Saddle River, NJ: Prentice Hall PTR

Rao, D.G. (2010). Introduction to Biochemical engineering. 2nd Ed.

Tata McGraw Hill Education Private Limited

Dutta, R. (2008). Fundamentals of Biochemical Engineering. New Delhi:

Ane Books India

Katoh, S. and Yoshida, F. (2009). Biochemical Engineering: A Textbook

for Engineers Chemists and Biologists. Weinheim: Wiley-VCH Verlag
GmbH & Co

Nielsen, J., Villadsen, J. and Gunner L. (2011). Bioreaction Engineering

Principles. 3rd Ed. New York: Springer Science+Bussiness Media

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