ENZYMES

Content:
1. History of Enzymes
2. The role of enzymes
3. Chemical reaction rates and the effects of catalysis
4. Enzymes as catalysts
5. The kinetics of enzyme catalysis
6. Enzyme inhibition
7. The regulation of enzyme activity
8. Coenzymes, vitamins and essential metals
9. The diversity of enzymatic functions
10. The regulation of enzyme activity

History of Enzymes

Friedrich Wilhelm Kuhne

 He coined the term “enzyme” in 1878\
 “biological catalysts” that had previously been called “ferments”
 The word itself means 'in yeast' from the Greek word “enzymos”
“en” means in
“zyme” means yeast or leaven
 Because of most recognizable reaction popularly known as alcohol fermentation by
zymase enzyme in yeast.

Number of enzymes per cell

 A bacterium like Escherichia coli has about 4228 proteins of which almost 1701 of
them are enzymes.
 Mammals have more than 10 times the number of proteins and enzymes found in E.
coli.
 The human body makes approx. 22 digestive enzymes.
Enzyme
 It is a protein molecule that functions as a catalyst to speed up chemical reactions
in living organisms.

Characteristics of Enzyme
1. Made of proteins (or RNA)
 They are actually globular proteins. Like all other proteins, enzymes are built with
amino acids structured in a long chain which folds to generate specifically ordered
three-dimensional structures. Each enzyme consists of between a hundred and up
to a million amino acids placed like pearls on a string. The structure and function
of an enzyme is decided by the order of the amino acids. The unique three-
dimensional structure of each enzyme determines the function of the enzyme. Even
slight changes in the sequence of the amino acids could have a distinct impact on
the three-dimensional structure and function of the enzyme. Some RNA molecules
called ribozymes can also be enzymes. These are usually found in the nuclear
region of cells and catalyze the splitting of RNA molecules.

2. Biological catalysts
 Speeds up chemical reactions. They catalyse nearly all the chemical reactions
taking place in the cells of the body.

3. Reusable
 Used in multiple reactions. It cannot be altered or consumed during the reaction.
They remain unchanged after each reaction and can therefore be reused.

4. Specific
 One type of enzyme for one type of reaction.

Naming of Enzyme SUBSTRATE ENZYMES PRODUCTS

lactose lactase glucose + galactose
 Usually ends in –ase maltose maltase glucose
 Identifies a reacting substance cellulose cellulase glucose
sucrase – reacts sucrose lipid lipase glycerol + fatty acid
lipase - reacts lipid starch amylase maltose
 Describes function of enzyme protein protease peptides + polypeptide
oxidase – catalyzes oxidation
hydrolase – catalyzes hydrolysis
 Common names of digestion
enzymes still use –in
pepsin, trypsin

Systemic Name
 Each enzyme is characterized by a code no. called Enzyme Code no. or EC
number and contain four figures (digits) separated by a dot.
EC m.n.o.p SUBSTRATE ENZYMES PRODUCTS
lactose lactase glucose + galactose
maltose maltase glucose
cellulose cellulase glucose
lipid lipase glycerol + fatty acid
starch amylase maltose
  First digit represents the class;
 Second digit stands for subclass;
 Third digit stands for sub-sub class or subgroup;
 Fourth digit gives the serial number of the particular enzymes in the list.
Example: EC 2.7.1.1.for hexokinase

Classification of Enzymes

Examples of biological enzymes

 Lipase — breaking down of fats (lipid)
 Protease — breaking down of proteins
 Cellulase — breaking down of fiber (cellulose)
 Amylase — breaking down of starch (amylose)
 Lactase — breaking down of dairy products (lactose)
 Sucrase — breaking down of sugar (sucrose)
 Maltase — breaking down of grains (maltose)

Mechanism of Enzyme Action

The basic mechanism of enzyme action is to catalyze the chemical reactions, which
begins with the binding of the substrate with the active site of the enzyme. This active site is a
specific area that combines with the substrate. After the enzyme and substrate bind it is called
an enzyme-substrate complex. The substrate may break apart or bond together to form a
product.
 The molecules that bind to the enzyme are called
substrates.
 The location where they bind (sit down) is called the
active site.

Enzymes match with a substrate that

fits like a puzzle at an active site.
 The basic mechanism of enzyme catalysis involving
enzyme substrate binding→ ES complex → EP complex→ enzyme + product

Example:
Maltase

Maltose is made up two glucose molecules bonded together. The maltase enzyme
is a protein that is perfectly shaped to accept a maltose molecule and break the bond. Then the
two glucose molecules are released. A single maltase enzyme can break in excess of 1,000
maltose bonds per second, and will only accept maltose molecules.

Catalytic Cycle of an Enzyme

Example:
Sucrase

Two Models of Enzyme Action

1. Lock-and-Key Model
 Proposed by Emil Fischer in 1894
 The active site has a rigid shape
 Only substrates with the matching shape can fit
 The substrate is a key to fits the lock of the
active site

In this model, the shape of the active site and substrate complement in such a way that
the substrate fits into the binding site perfectly. There’s some truth in the lock and key model
in that enzymes do have active sites, which need to be filled with a substrate and interact with
the substrate through non-covalent interactions. The problem with this model is that if the
substrate fitted the enzyme perfectly, catalysis would be hampered. This is because the enzyme
needs to bring about a change to the substrate and not just to bind it. The Lock and Key model
explains that the enzyme needs to bind substrate, but once the reaction progresses to the
transition state and product formation, the active site would not be able to accommodate this
change. This is the paradox of how enzymes work, they need to be able to bind specifically to
the substrate, but they also need to be able to turn the substrate into something else (product),
which means those two things are at odds with each other. So the enzyme needs to bind the
substrate slightly imperfectly in order to be able to turn it over, that is, convert it to the product.
This is an older model, however, and does not work for all enzymes.
2. Induced-Fit Model
 Proposed by Danial Kosh Lan in 1958.
 The active site is flexible, not rigid.
 The shapes of the enzyme, active site, and substrate adjust to maximize the fit, which
improves catalysis.
 There is a greater range of substrate specificity.

 Refine the original lock-and-key model of enzyme actions. They discovered that the
binding of a substrate often leads to a large conformational change in the enzyme, as
well as to changes in the structure of the substrate or substrates. The current theory,
known as the induced-fit model, says that enzymes can undergo a change in
conformation when they bind substrate molecules, and the active site has a shape
complementary to that of the substrate only after the substrate is bound.
 This model is more consistent with a wider range of enzymes.
In the Induced Fit model, the enzyme active site forms in response to substrate binding.
Initially the active site is not perfect, but upon binding, it is able to move, which puts the active
site under strain. This strain is then able to elicit the energy that’s required for the reaction to
occur by stabilizing the transition state and not just binding of the substrate. The enzyme carries
out its work by inducing the substrate to take up a transition state on the path to the required
product.

Enzymes as catalysts in chemical reactions

 Chemical reactions occur when molecules interact and chemical bonds between
them are formed or broken.
 The rate of reaction or reaction rate is a measurement of how fast a reaction occurs.

Enzymes speed the reaction, or allow it

to occur at lower energy levels and, once
the reaction is complete, they are again
available. Most often, we look at how
quickly products are being made or how
quickly substrates are being used up in
order to determine the rate of reaction.
Adding an enzyme or other catalyst to a
reaction will speed up its rate by
decreasing activation energy.
Activation Energy
 It is the energy required for a chemical reaction to occur.
 Enzymes lower the activation energy barrier of a reaction because they
are catalysts. This means that reactions can progress much faster than they can
without a catalyst. In other words, the reaction speeds up.

 Enough energy has to be added to the reactants to get a reaction to start. It's like pushing
a car up a hill: you have to keep adding more and more energy, but once you're at the
top of the hill, the car (or the reaction) can begin to roll on its own.
 It takes time to get enough energy to start a reaction. Enzymes shorten this time by
reducing the activation energy required for a reaction to begin.

Example:
Catecholase
 Catecholase is present in most fruits and vegetables.
 It is the enzyme that facilitates the browning of cut or
bruised fruits and vegetables by catalyzing the following
reaction:

(catecholase)
Catechol + oxygen ------------ polyphenol
colorless substrate brown product

Factors Affecting Enzyme Activity

1. Temperature: Raising temperature generally

speeds up a reaction, and lowering temperature
slows down a reaction. However, extreme high
temperatures can cause an enzyme to lose its
shape (denature) and stop working.

2. pH: Each enzyme has an optimum pH range.

Changing the pH outside of this range will slow
enzyme activity. Extreme pH values can cause
enzymes to denature.
Enzymes require an optimum temperature and pH for
their action. The temperature or pH at which a compound
shows its maximum activity is called optimum temperature or optimum pH, respectively. As
mentioned earlier, enzymes are protein compounds. A temperature or pH more than optimum
may alter the molecular structure of the enzymes. Generally, an optimum pH for enzymes is
considered to be ranging between 5 and 7.
Enzymes can be denatured - they change shape so much that they are no longer effective.
High temp or pH can cause denaturation by breaking the H-bonds of the protein 4D structure.
When food is cooked, some of its proteins become denatured. This is why boiled eggs become
hard and cooked meat becomes firm.

Denaturation
 the change of folding structure of a protein (and thus of physical properties) caused by
heating, changes in pH, or exposure to certain chemicals
 Temperatures far above the normal range denature enzymes. (This is why very high
fevers are so dangerous. They can cook the body’s proteins.)
 Most enzymes work best near neutral pH (6 to 8).
Example: Irreversible egg protein denaturation caused by high temperature (while cooking it).

3. Enzyme/Substrate Concentration

Enzyme concentration: Increasing enzyme

concentration will speed up the reaction, as
long as there is substrate available to bind to.
Once all of the substrate is bound, the reaction
will no longer speed up, since there will be
nothing for additional enzymes to bind to.

Substrate concentration: Increasing substrate

concentration also increases the rate of reaction
to a certain point. Once all of the enzymes have
bound, any substrate increase will have no effect
on the rate of reaction, as the available enzymes
will be saturated and working at their maximum
rate.

Kinetics of Enzyme Catalysis

Enzyme catalysis
 It is an increase in the rate of reaction with the help of enzyme (as catalyst)
Michaelis–Menten kinetics
 is one of the simplest and best-known
models of enzyme kinetics. It is named
after Leonor Michaelis and Maud
Menten. This model is based on the
single substrate reaction:
  If the initial rate of the reaction is
measured over a range of substrate
concentrations (denoted as [S]), the
reaction rate ( increases / decreases ) as
[S] increases.
 As [S] gets higher, the enzyme
becomes saturated with substrate and the
rate reaches Vmax, the enzyme's
maximum rate.

4. Presence of Inhibitors
Inhibitors
 substances which alter the catalytic action of the enzyme and consequently slow
down, or in some cases, stop catalysis. This effect may
be permanent or temporary.

Types of Inhibitors:
a. Competitive Inhibitor
 An inhibitor may bind to an enzyme
and block binding of the substrate, for
example, by attaching to the active
site. The inhibitor “competes” with the
substrate for the enzyme. That is, only
the inhibitor or the substrate can be
bound at a given moment.
 Chemicals that resemble an enzyme’s
normal substrate and compete with it
for the active site.
 Reversible depending on concentration
of inhibitor and substrate.
Example: The drug Antabuse is used to help alcoholics quit drinking. Antabuse inhibits
aldehyde oxidase, resulting in the accumulation of acetaldehyde during the metabolism of
alcohol. Elevated acetaldehyde levels cause symptoms of nausea and vomiting.

b. Noncompetitive Inhibitor
 An inhibitor that doesn't block the
substrate from binding to the active site.
Instead, it attaches at another site called
“allosteric site” and blocks the enzyme
from doing its job. This inhibition is said
to be "noncompetitive" because the
inhibitor and substrate can both be
bound at the same time.
 Do not enter active site, but bind to
another part of the enzyme, causing the
enzyme & active site to change shape.
 Usually reversible, depending on
concentration of inhibitor & substrate.
Example: You may know that compounds containing heavy metals such as lead, mercury,
copper or silver are poisonous. This is because ions of these metals are non-competitive
inhibitors for several enzymes.

Regulatory Molecules

Allosteric regulation, is just any form of regulation where the regulatory molecule (an
activator or inhibitor) binds to an enzyme someplace other than the active site. The place where
the regulator binds is called the allosteric site.
Inhibitors – decrease the activity of an enzyme
Activators – Increase the activity of an enzyme
Enzyme Inhibitors as Medicines
Some Enzyme Inhibitors can be
used as Medicines in the treatment
of conditions. For example, infection by
viruses can be treated by Inhibitors to the
viral enzyme Protease, often competitive
Inhibitors. This means that viruses cannot
build new protein coats and therefore
cannot replicate.
Penicillin works by inhibiting a
bacterial enzyme that is responsible for
forming cross-links in bacteria cell walls.
This therefore halts reproduction.

Coenzymes, Vitamins and essential metals

Many enzymes are simple proteins consisting

entirely of one or more amino acid chains.
Other enzymes contain a nonprotein
component called a cofactor that is necessary
for the enzyme’s proper functioning. There
are two types of cofactors: inorganic ions
[e.g., zinc or Cu(I) ions] and organic
molecules known as coenzymes.
Most coenzymes are vitamins or are derived
from vitamins.
Example: DNA polymerase requires a Zinc
ion (𝑍𝑛2+ ) to build DNA molecules.

Function of Coenzyme
 A coenzyme prepares the active site for catalytic activity.
Metal Ions as Cofactors
Many active enzymes require a
metal ion. For example, Zn2+ , a cofactor
for carboxypeptidase, stabilizes the
carbonyl oxygen during the hydrolysis of a
peptide bond.

Vitamins-derived Coenzymes
Vitamins are required for coenzyme synthesis and must be supplied in the diet.
 Lack of particular vitamins causes disease.

Vitamins C and E, as well as the provitamin β-carotene can act as antioxidants in the
body. Antioxidants prevent damage from free radicals, which are molecules that are highly
reactive because they have unpaired electrons. Free radicals are formed not only through
metabolic reactions involving oxygen but also by such environmental factors as radiation and
pollution.
Because organisms differ in their synthetic abilities, a substance that is a vitamin for
one species may not be so for another. Over the past 100 years, scientists have identified and
isolated 13 vitamins required in the human diet and have divided them into two broad
categories: the fat-soluble vitamins, which include vitamins A, D, E, and K, and the water-
soluble vitamins, which are the B complex vitamins and vitamin C. All fat-soluble vitamins
contain a high proportion of hydrocarbon structural components. There are one or two oxygen
atoms present, but the compounds as a whole are nonpolar. In contrast, water-soluble vitamins
contain large numbers of electronegative oxygen and nitrogen atoms, which can engage in
hydrogen bonding with water. Most water-soluble vitamins act as coenzymes or are required
for the synthesis of coenzymes.

Two catagories of vitamins:

1. Water-soluble (B vitamins and Vitamin C)
 Required daily in diet
 Excess excreted in urine
2. Lipid-soluble (Vitamins A,D,E,K)
 Intake must be limited
 Stored in fat
Functions and Applications of Enzymes

Enzymes are essential for respiration, digesting food, muscle and nerve function, among
thousands of other roles.
 Some enzymes help to break down
large nutrient molecules, such as
proteins, fats, and carbohydrates, into
smaller molecules. This process occurs
during the digestion of foodstuffs in the
stomach and intestines of animals.

 Cleaning Products
 Many biological washing powders contain enzymes to help with the removal of
stains. The enzyme may be a protease to break down protein stains or a lipase
to break down fat an oil (lipids) stains.
 Beverages
 Alcoholic beverages generated by fermentation vary a lot based on many
factors. Based on the type of the plant’s product, which is to be used and the
type of the enzyme applied, the fermented product varies.
 For example, grapes, honey, hops, wheat, cassava roots, and potatoes depending
upon the materials available. Beers, wines and other drinks are produced from
plant fermentation.

 Food Products
 Bread can be considered as the finest example of fermentation in our everyday
life.
 A small proportion of yeast and sugar is mixed with the batter for making bread.
Then one can observe that the bread gets puffed up as a result of fermentation
of the sugar by the enzyme action in yeast, which leads to the formation of
carbon dioxide gas. This process gives the texture to the bread, which would be
missing in the absence of the fermentation process.
References:
 https://byjus.com/biology/enzymes/
 https://www.pinterest.ph/pin/486951778443313272/
 https://sites.duke.edu/apep/module-1-gender-matters/biology-and-chemistry-
connections/enzymes-as-
catalysts/#:~:text=Enzymes%20are%20proteins%20that%20have,chemical%20reacti
ons%20that%20they%20accelerate.
 https://www.ncbi.nlm.nih.gov/books/NBK9921/
 https://www.slideshare.net/akeeeee/enzyme-cofactorsandvitamins
 https://en.wikipedia.org/wiki/Competitive_inhibition
 https://courses.lumenlearning.com/boundless-biology/chapter/enzymes/
 https://www.news-medical.net/life-sciences/What-is-an-Enzyme-
Cofactor.aspxhttps://slideplayer.com/slide/11374568/