Enzyme

Introduction

The living cell is the site of tremendous biochemical activity called metabolism. This is the process of chemical
and physical change which goes on continually in the living organism. Build-up of new tissue, replacement of old
tissue, conversion of food to energy, disposal of waste materials, reproduction - all the activities that we
characterize as "life." the greatest majority of these biochemical reactions do not take place spontaneously. The
phenomenon of catalysis makes possible biochemical reactions necessary for all life processes. Catalysis is
defined as the acceleration of a chemical reaction by some substance which itself undergoes no permanent
chemical change. The catalysts of biochemical reactions are enzymes and are responsible for bringing about
almost all of the chemical reactions in living organisms. Without enzymes, these reactions take place at a rate far
too slow for the pace of metabolism.

The existence of enzymes has been known for well over a century. Some of the earliest studies were performed
in 1835 by the swedish chemist jon jakob berzelius who termed their chemical action catalytic. It was not until
1926, however, that the first enzyme was obtained in pure form, a feat accomplished by james b. Sumner of
cornell university. Sumner was able to isolate and crystallize the enzyme urease from the jack bean. His work
was to earn him the 1947 nobel prize.

What is an enzyme? How do they work?

An enzyme is a protein that functions as a catalyst to speed up a chemical reaction in the body; it is not used up
in the chemical reaction, rather it is recycled and used over and over again. All enzymes are proteins. Enzymes
are biological catalyst.

Characteristics of enzymes:

1. Enzymes do not make anything happen that couldn’t happen on its own, just makes it happen faster.
2. Enzymes are not used up in reactions. They can be used over and over again!
3. Enzymes are only needed in small amounts.
4. Each enzyme is highly selective about its substrate.
5. Enzymes chemically recognize, bind and modify substrates.

Enzyme action – can occur two ways:

1. Lock and key model – the substrate molecule has a specific 3-

dimensional shape that allows it to fit into the specific 3-dimensional shape
of an enzyme’s active site. Both enzyme and substrate already exist in these
specific 3-dimensional shapes.
2. Induced fit model – an interaction between the enzyme and substrate
induces or changes the shape of the molecules to produce a suitable fit.
What environmental factors can affect an enzyme’s function?

1. Temperature:

The big idea: enzymes function optimally at certain temperatures.

As temperature increases, kinetic energy increases and molecules are moving more, increasing the likelihood
that enzyme and substrate molecules will “bump into” each other, bind, and react. Therefore, initially enzyme
reaction rate increases with an increase in temperature. The optimal temperature for an enzyme is the
temperature at which the enzyme “works best,” and the rate of chemical reaction is highest. The “optimal
temperature” for most of the enzymes in your body is ~98.6 degrees f (also known as ~37 degrees c).

Trends:

 At extremely cold temperatures enzymes work slowly or not at all.

 Warm temperatures generally increase relative rate (speed) of an enzyme.
 Extremely high temperatures inactivate or “denature” the enzyme.

2. Ph (a measure of acidity)

The big idea: enzymes function optimally at a certain ph.

Changes in ph can make and break chemical bonds within the enzyme, changing the shape of the enzyme and,
therefore, its effectiveness. If the ph is too low (too acidic) or too high (too basic), the enzyme becomes
“denatured”: the chemical bonds within the enzyme are rearranged and the enzyme becomes misfolded. As the
enzyme’s shape changes, the 3- dimensional shape of its active site changes, and the active site cannot bind to
the substrate anymore. Thus, the enzyme cannot function anymore and the reaction rate decreases sharply.

Trends:

 Acidic ph: <ph7

 Neutral ph: =ph7
 Basic ph: >ph7
 Optimal ph: the ph at which the enzyme “works best,” and the chemical reaction goes fastest
 The “optimal ph” for most of the enzymes in your body is ~ph8
 However there are exceptions, such as the digestive enzymes of your stomach, which function in an
environment of ~ph3-4

3. Concentration of Enzyme or Substrate

When enzyme concentration is low, the reaction is slower. As enzyme concentration increases, the reaction is
faster up to a point when the amount of substrate available becomes limiting. Similarly, when substrate
concentration is low, the reaction is slower. As substrate concentration increases, the reaction is faster up to a
point when the amount of enzyme available becomes limiting.
Enzyme kinetics

The study of reaction rates and how they change in response to changes in experimental parameters in known
as Kinetics. Kinetics is that branch of enzymology that deals with the factors that affect the rate of enzyme
catalyzed reactions.

Factors affecting Enzyme Reaction Velocity:

1. Enzyme concentration.
2. Substrate concentration.
3. Temperature
4. Ph.
5. Activators.
6. Inhibitors

In a mathematical description of enzyme action developed by Leonor Michaelis and Maud Menten in 1913, two
constants, Vmax and Km, play an important role. These constants are important to know, both to understand
enzyme activity on the macroscale and to understand the effects of different types of enzyme inhibitors.

Maximal Velocity (Vmax): Increasing the substrate concentration indefinitely does not increase the rate of an
enzyme-catalyzed reaction beyond a certain point. This point is reached when there are enough substrate
molecules to completely fill (saturate) the enzyme's active sites. The maximal velocity, or Vmax, is the rate of the
reaction under these conditions. Vmax reflects how fast the enzyme can catalyze the reaction.

Michaelis Constant (Km): Enzymes have varying tendencies to bind their substrates (affinities). An enzyme's Km
describes the substrate concentration at which half the enzyme's active sites are occupied by substrate. A high
Km means a lot of substrate must be present to saturate the enzyme, meaning the enzyme has low affinity for
the substrate. On the other hand, a low Km means only a small amount of substrate is needed to saturate the
enzyme, indicating a high affinity for substrate.

Michaelis – Menten Plot

Two 20th century scientists, Leonor Michaelis and Maud Leonora Menten, proposed the model known as
Michaelis-Menten Kinetics to account for enzymatic dynamics. The model serves to explain how an enzyme can
cause kinetic rate enhancement of a reaction and explains how reaction rates depends on the concentration of
enzyme and substrate. The Michaelis-Menten model (1) is the one of the simplest and best-known approaches
to enzyme kinetics. It takes the form of an equation relating reaction velocity to substrate concentration for a
system where a substrate S binds reversibly to an enzyme E to form an enzyme-substrate complex ES, which
then reacts irreversibly to generate a product P and to regenerate the free enzyme E. This system can be
represented schematically as follows:

The Michaelis-Menten equation for this system is:

 Here, Vmax represents the maximum velocity achieved by the system,
at maximum (saturating) substrate concentrations. KM (the Michaelis
constant; sometimes represented as KS instead) is the substrate
concentration at which the reaction velocity is 50% of the Vmax. [S] is
the concentration of the substrate S.

This is a plot of the Michaelis-Menten equation’s predicted reaction

velocity as a function of substrate concentration, with the significance
of the kinetic parameters Vmax and KM graphically depicted.

Lineweaver–Burk plot

In biochemistry, the Lineweaver–Burk plot (or double reciprocal plot) is a graphical representation of the
Lineweaver–Burk equation of enzyme kinetics, described by Hans Lineweaver and Dean Burk
in 1934.

The plot provides a useful graphical method for analysis of the Michaelis–Menten equation:

The Lineweaver–Burk plot was widely used to determine important terms in enzyme kinetics, such as Km and
Vmax, before the wide availability of powerful computers and non-linear regression software. The y-intercept of
such a graph is equivalent to the inverse of Vmax; the x-intercept of the graph represents −1/Km. It also gives a
quick, visual impression of the different forms of enzyme inhibition. When used for determining the type of
enzyme inhibition, the Lineweaver–Burk plot can distinguish competitive, non-competitive and uncompetitive
inhibitors. Competitive inhibitors have the same y-intercept as uninhibited enzyme (since Vmax is unaffected by
competitive inhibitors the inverse of Vmax also doesn't change) but there are different slopes and x-intercepts
between the two data sets. Non-competitive inhibition produces plots with the same x-intercept as uninhibited
enzyme (Km is unaffected) but different slopes and y-intercepts. Uncompetitive inhibition causes different
intercepts on both the y- and x-axes.
References:

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