Enzymes are proteins that act as catalysts, helping speed up chemical reactions in living cells.

●​ Without enzymes, most reactions in cells would be too slow to sustain life.​

●​ Almost all enzymes are globular proteins, except for a few RNA molecules that can also
act as catalysts.

Enzymes (Simple Explanation)

●​ Enzymes are special proteins that help speed up chemical reactions in the body.​

●​ Some enzymes can make reactions happen much faster—even a billion times quicker
than they would on their own. (1020 times)​

●​ They are important because:​

○​ They work very fast to help the body function properly.​

○​ They are specific, meaning each enzyme only works for one type of reaction.​

○​ They can be controlled, so the body can turn them on or off as needed.​

Without enzymes, many processes in the body would be too slow to keep us alive.

Catalytic Efficiency

●​ Catalysts make chemical reactions happen faster without being used up.​

●​ They participate in the reaction but stay the same and can be used again and again.​

●​ Enzymes work the same way as other catalysts by lowering the energy needed for a
reaction, helping it reach a balanced state more quickly.​

Catalytic Efficiency

●​ Enzymes help carry out important chemical reactions in the body, such as breaking
down esters, oxidizing alcohols, and forming amides.​

●​ Unlike chemical reactions in a test tube, enzymes allow these processes to happen
under mild conditions of pH and temperature.​
 ●​ What might take hours or weeks in a laboratory can happen in seconds with enzymes.​

●​ The enzyme carbonic anhydrase speeds up the removal of carbon dioxide by

combining it with water to form carbonic acid much faster than it would without the
enzyme.​

Further explanation:

Carbonic Anhydrase: How It Helps Remove CO₂

●​ Your body makes carbon dioxide (CO₂) when it uses energy.​

●​ Too much CO₂ in the blood is dangerous, so your body needs to get rid of it fast.​

●​ Carbonic anhydrase is an enzyme that helps CO₂ mix with water to form a new
substance called carbonic acid.​

●​ This process happens much faster with the enzyme than without it.​

●​ In the lungs, the reaction reverses, turning carbonic acid back into CO₂, which you
breathe out.​

Without carbonic anhydrase, getting rid of CO₂ would be too slow, and your body wouldn't work
properly.

Specificity

●​ Enzymes are highly specific, meaning they only work on certain reactions and specific
substances.​

●​ Unlike strong acids, which can break down many different compounds, enzymes are
more selective.​

●​ For example, the enzyme urease only breaks down urea, a single type of amide, while
strong acids can break down many amides and esters.​

Specificity of Enzymes

Enzymes work on specific substances, but their level of specificity can vary:
 ●​ Absolute specificity → The enzyme only reacts with one exact substance and nothing
else.​

○​ Example: An enzyme that only works on a single molecule.​

●​ Relative specificity → The enzyme works on a group of similar substances that

share a common structure.​

○​ Example:​

■​ Lipases break down fats (lipids).​

■​ Proteases break down proteins.​

■​ Phosphatases break down phosphate compounds.​

●​ Stereochemical specificity → The enzyme only reacts with one version of a

molecule when two mirror-image forms exist.​

○​ Example: D-amino acid oxidase only breaks down D-amino acids and does not
work on L-amino acids.​

This means some enzymes are very strict and only work on one substance, while others can
work on a group of similar substances.

Regulation of Enzymes

●​ Enzymes can be controlled to regulate their activity.​

●​ Even though many chemical reactions could happen in a cell, only a few actually occur
because enzymes choose which ones take place.​

●​ The cell controls how fast these reactions happen and how much of a product is made
by regulating enzyme activity.​

Enzyme Nomenclature

●​ Early enzymes were named with "-in" at the end to show they were proteins (e.g.,
pepsin, trypsin, chymotrypsin).​
 ●​ Since there are now many known enzymes, a systematic naming system called the
Enzyme Commission (EC) system is used.​

●​ Enzymes are divided into six major classes based on the type of reaction they help
with.​

●​ Each enzyme has a specific name that describes:​

○​ The substrate (the substance it acts on).​

○​ The functional group it affects.​

○​ The reaction type it catalyzes.​

●​ All EC enzyme names end with "-ase".​

Difference Between Substrate and Cofactor

●​ Substrate → The substance that the enzyme acts on and changes into a new
product.​

○​ Example: Amylase (enzyme) breaks down starch (substrate) into sugar.​

●​ Cofactor → A helper molecule that some enzymes need to function properly.​

○​ Cofactors can be metal ions (like zinc or iron) or organic molecules

(coenzymes like vitamins).​

○​ Example: The enzyme carbonic anhydrase needs zinc as a cofactor to speed

up the removal of CO₂ from the body.​

Main Difference

●​ Substrate = What the enzyme works on.​

●​ Cofactor = What helps the enzyme work.

EXAMPLE AGAIN :​
​
For carbonic anhydrase, the substrates are carbon dioxide (CO₂) and water (H₂O). The
enzyme speeds up their conversion into carbonic acid (H₂CO₃), which later breaks down into
CO₂ and water for exhalation.
Zinc (Zn²⁺) acts as a cofactor to help the enzyme function properly.

Enzyme Cofactors

●​ Some enzymes need non-protein molecules or metal ions to function properly. These
are called prosthetic groups if they are tightly bound to the enzyme.​

●​ If the non-protein component is loosely attached, it is called a cofactor.​

Types of Cofactors:

1.​ Coenzymes → Organic molecules that help enzymes (e.g., vitamins).​

2.​ Inorganic ions → Usually metal ions like Mg²⁺, Zn²⁺, or Fe²⁺.​

The protein part of the enzyme (without the cofactor) is called an apoenzyme.

NOTE: APOENZYME + COFACTOR = ACTIVE ENZYME

Enzymatic Action

1.​ Enzymes work on specific substances by interacting with them at a special area
called the active site.​

2.​ When the substrate (the substance the enzyme acts on) binds to the active site, it
forms an enzyme-substrate (ES) complex.​

3.​ The enzyme helps break and form chemical bonds, changing the substrate into a
new product.​

4.​ After the reaction, the product is released, and the enzyme is ready to work on
another substrate.​

EXAMPLE:

Enzymatic Action (Example: Amylase and Starch)

Amylase is an enzyme that breaks down starch into smaller sugars. When starch (the substrate)
binds to the active site of amylase, they form an enzyme-substrate (ES) complex. At the active
site, amylase helps break the chemical bonds in starch, converting it into maltose and other
simple sugars. Once the reaction is complete, the sugar molecules (products) are released, and
amylase is free to bind to another starch molecule and repeat the process.
Lock and Key Theory

The lock-and-key theory explains why enzymes are highly specific. The enzyme's active site is
like a lock, and only a substrate with the right shape and size can fit into it, forming an
enzyme-substrate (ES) complex.

A limitation of this theory is that it assumes enzymes have rigid shapes. However, research
suggests that enzymes are somewhat flexible, meaning their shape can slightly adjust to fit the
substrate better.

Enzymatic Activity

●​ Enzyme activity refers to how well an enzyme speeds up a reaction.​

●​ The turnover number is the number of substrate molecules processed by a single

enzyme per minute.​

○​ Carbonic anhydrase has one of the highest turnover numbers, acting on 36

million molecules per minute.​

○​ Most enzymes process around 1,000 molecules per minute.​

Enzyme Assays

●​ Enzyme assays are experiments used to measure enzyme activity.​

●​ In clinical labs, enzyme assays help analyze blood enzymes.​

●​ These tests often measure enzyme activity by:​

○​ Observing color changes in the product or substrate.​

○​ Tracking pH changes over time for reactions involving H⁺ ions.​

Factors Affecting Enzyme Activity

Enzyme Concentration

●​ The amount of enzyme ([E]) present affects the reaction rate.​

●​ More enzymes mean more reactions, so doubling the enzyme concentration doubles
the reaction rate.​
 ●​ The relationship between enzyme concentration and reaction rate forms a straight-line
graph.​

Substrate Concentration

●​ The amount of substrate ([S]) also affects reaction speed.​

●​ More substrate increases the reaction rate until the enzyme becomes saturated.​

●​ Once saturation is reached, the reaction hits a maximum speed (Vmax) and cannot go
any faster.​

Temperature

●​ Higher temperatures generally increase enzyme activity—up to a point.​

●​ Too much heat denatures the enzyme, stopping its function.​

●​ Most enzymes work best between 25°C and 40°C; outside this range, activity
decreases.​

pH Levels

●​ pH changes can alter enzyme structure and function.​

●​ Extreme pH levels denature enzymes, making them ineffective.​

●​ Most enzymes work best at pH 7, but some, like pepsin in the stomach, function better
in acidic conditions.​

Enzyme Inhibition Explained in Detail

Enzyme inhibition occurs when a substance (inhibitor) reduces or stops an enzyme's ability to
catalyze a reaction. This can happen in two main ways: irreversible inhibition and reversible
inhibition.

Irreversible Inhibition
Irreversible inhibitors permanently disable enzymes by forming strong covalent bonds with
specific functional groups. This destroys the enzyme’s activity, and the enzyme cannot
function again.

Examples of Irreversible Inhibitors:

1. Cyanide Poisoning (CN⁻)

●​ Cyanide binds permanently to the enzyme cytochrome oxidase, which is necessary

for cellular respiration (the process of producing energy in cells).​

●​ Without this enzyme, cells cannot produce ATP (energy), leading to cell death.​

●​ Result: Death occurs within minutes due to lack of energy.​

●​ Antidote: Sodium thiosulfate converts cyanide into thiocyanate, which does not block
cytochrome oxidase and can be safely excreted.​

2. Heavy Metal Poisoning (Lead & Mercury)

●​ Lead (Pb²⁺) and Mercury (Hg²⁺) bind to –SH groups (sulfhydryl groups) on enzymes,
preventing them from working properly.​

●​ This can lead to protein denaturation (loss of enzyme structure), making the enzyme
completely useless.​

●​ Result: Can cause permanent neurological damage (brain damage, memory loss,
etc.).​

●​ Treatment: Chelating agents (such as EDTA) bind tightly to heavy metal ions and help
remove them through urine.​

Antibiotics as Enzyme Inhibitors

Certain antibiotics kill bacteria by inhibiting their enzymes, preventing them from functioning.

Examples of Antibiotics that Act as Enzyme Inhibitors:

1.​ Sulfa Drugs (Discovered by Gerhard Domagk in 1935)​

 ○​ These block enzymes that bacteria need to make folic acid, an essential nutrient
for their survival.​

○​ Humans do not make folic acid (we get it from food), so sulfa drugs only affect
bacteria and not human cells.​

2.​ Penicillin (Discovered by Alexander Fleming in 1928)​

○​ Blocks the enzyme transpeptidase, which bacteria use to build cell walls.​

○​ Without this enzyme, bacteria cannot form strong cell walls, causing them to
burst and die.​

Reversible Inhibition
Unlike irreversible inhibitors, reversible inhibitors bind to enzymes temporarily and can be
removed by shifting equilibrium, allowing the enzyme to function again.

Enzyme Inhibition — Competitive Inhibitors

●​ Competitive inhibitors bind to the active site of an enzyme, preventing the normal
substrate from attaching.​

●​ These inhibitors have a similar shape to the substrate, allowing them to compete for the
active site.​

Competitive Inhibition in Action

●​ There are two equilibria happening: the enzyme binding to either the substrate or the
inhibitor.​

●​ This inhibition can be reversed by:​

○​ Increasing substrate concentration, so more substrate molecules outcompete

the inhibitor.​

○​ Decreasing inhibitor concentration, reducing competition for the active site.​

Enzyme Inhibition — Noncompetitive Inhibitors

●​ Noncompetitive inhibitors do not bind to the active site.​

●​ Instead, they attach to a different site on the enzyme, changing its shape and making
the active site ineffective.​

Noncompetitive Inhibition in Action

●​ Unlike competitive inhibitors, noncompetitive inhibitors do not resemble the substrate

and cannot be outcompeted by increasing substrate concentration.​

●​ Since the enzyme’s shape is altered, the substrate no longer fits properly, stopping the
reaction.​

Enzyme Regulation

Enzymes control biochemical reactions in living organisms and must be regulated to respond to
changes in cellular conditions. Regulation occurs through:​
✔ Activation of zymogens​
✔ Allosteric regulation​
✔ Genetic control

Activation of Zymogens

●​ Zymogens (proenzymes) are inactive enzyme precursors.​

●​ Some enzymes, if active too soon, could damage the cell, so they are stored in an
inactive form.​

●​ When needed, the zymogen is released and activated in the right location.​

●​ Activation happens by cleaving peptide bonds in the zymogen.​

●​ Examples: Pepsin, trypsin, chymotrypsin (digestive enzymes) and blood clotting

enzymes.​

Allosteric Regulation

●​ Some molecules, called modulators, can bind to enzymes and change their 3D shape.​
 ●​ This change affects how well the enzyme works:​

○​ Activators make the enzyme work better.​

○​ Inhibitors make the enzyme work worse (similar to noncompetitive inhibitors).​

●​ Enzymes that have these special binding sites for modulators are called allosteric
enzymes.​

●​ These enzymes help control important cell processes, adjusting their activity based
on what the cell needs.​

Feedback Inhibition

●​ This is a way for cells to self-regulate enzyme activity.​

●​ When too much of a final product builds up, it inhibits an earlier enzyme in the reaction.​

●​ This prevents waste and keeps the reaction balanced.​

●​ Example: Isoleucine Synthesis​

○​ The enzyme threonine deaminase starts the process of making isoleucine.​

○​ When isoleucine levels get too high, it binds to threonine deaminase, stopping
more isoleucine from being made.​

○​ Once isoleucine levels drop, the enzyme becomes active again, allowing more
to be produced.​

Genetic Control

●​ Cells can turn enzyme production on or off depending on their needs.​

●​ This ensures that enzymes are only made when they are actually needed.​

●​ Enzyme induction happens when a cell starts producing an enzyme in response to

environmental changes.​

Example: β-Galactosidase in E. coli

 ●​ The β-galactosidase enzyme breaks down lactose into simple sugars.​

●​ When there’s no lactose, the cell produces very little of this enzyme.​

●​ When lactose is present, the cell starts making a lot of β-galactosidase.​

●​ If lactose is removed again, the cell stops making the enzyme to save energy.​

This system helps cells adapt to their environment by only making enzymes when they are
actually needed.

Enzymes in Clinical Diagnosis

●​ Some enzymes should only be inside cells, so if they appear in the blood, it may mean
tissue damage.​

●​ Doctors check enzyme levels in blood to detect diseases in the heart, liver, pancreas,
prostate, and bones.​

Isoenzymes

●​ Isoenzymes are different versions of the same enzyme found in different body parts.​

●​ Example: LDH (Lactate Dehydrogenase) has five types, depending on whether it's
from the heart (H) or muscles (M).​

●​ LDH levels help diagnose diseases:​

○​ High LDH1 & LDH2 → Possible heart attack.​

○​ High LDH5 → Possible liver damage.​