Enzymes in the Body

Enzymes are molecules in the human body that help speed up chemical reactions. Some enzymes help
with the digestion of food in the body, but there are thousands of different kinds of enzymes, each
designed to help with a specific reaction. For example, while you might use hydrogen peroxide externally,
when hydrogen peroxide forms inside the body, it is harmful to cells. Luckily, there is an enzyme that
helps neutralize oxidative compounds like hydrogen peroxide.

The enzyme catalase helps protect the body from oxidative cell damage by breaking down hydrogen
peroxide to water and oxygen. An enzyme has an active site to which specific compounds attach. The
molecules in the compound are referred to as substrates. Once the substrates attach to the enzyme, the
chemical reaction is sped up, the reaction takes place, and the reaction products are released. In the
case of hydrogen peroxide, water and oxygen are released. Once the reaction is complete, the enzyme is
available again for another substrate, and the process repeats.

Students can see how this process works with a hands-on science activity that simulates the function of
catalase enzymes in the body. Using hydrogen peroxide and yeast (which contains catalase), students
are guided in a hands-on experiment with a control and sample solutions containing varying amounts of
hydrogen peroxide. When hydrogen peroxide breaks down into oxygen and water, you can see the
release of oxygen in the formation of bubbles (or foam). Observing and comparing the visible changes
that occur with each sample solution, students can better visualize the enzyme activity that happens in
the body.

The Exploring Enzymes activity contains the directions for this family-friendly science activity.

Making Connections
Students interested in the function of enzymes, may also enjoy science projects and activities like these:

 A Juicy Project: Extracting Apple Juice with Pectinase

 Enzyme-Catalyzed Reactions-- What Affects Their Rates?
 I Love Ice Cream, But It Doesn't Love Me: Understanding Lactose Intolerance
 Liver Stinks!
 The Liver: Helping Enzymes Help You!
 Which Fruits Can Ruin Your Gelatin Dessert?

(https://www.sciencebuddies.org/blog/enzymes-foam-and-hydrogen-peroxide)

Enzymes: The Little Molecules That Bake Bread

By Emily Buehler on September 28, 2012

When I first began making bread, the science involved was always in the back
of my mind. I had an idea of what occurred—my diagram for the chemical
reactions in dough looked something like this:
When I started preparing a manual for a bread-making class, however, I really
began to wonder about the details. Is the sugar for fermentation part of flour?
How exactly does the yeast process this sugar? Do all the complex flavors of
bread really come from one organic molecule, ethanol? Numerous trips to the
university libraries helped me understand the enzymes involved in making the
dough.

When I realized that flour contains a very small amount of sugar, only one to
two percent, I thought, “Wait a minute, how is that possible? That’s not
enough to make dough rise.” Then I figured out that the starch in flour
provides most of the sugar for fermentation, and the starch must be broken
down into sugar before it can be fermented. This breakdown is the work of
enzymes.

An enzyme is defined as a large molecule, usually a protein, that catalyzes a

biological reaction. This means that the enzyme speeds up the reaction by
reducing whatever energy barrier is preventing the reaction from happening
quickly and easily.

When two molecules bump into each other, there is a chance they will react to
form new molecules. Sometimes this happens easily—the two molecules each
have an unstable site, for example, and when they bump, a bond forms
between the sites, creating a new, stable molecule. In other cases, however,
bonds in the reacting molecules must break (which requires energy) before
new bonds can form. The amount of energy needed to break the old bonds is
the energy barrier to the reaction. This is represented by the solid line in the
diagram below.

One way to increase the speed of a reaction is to heat it up. Hotter molecules
move faster; they possess more energy. When two of them collide, there is a
greater chance that the necessary bonds will break and reaction will occur. If
more molecules possess the energy needed to get over the barrier, more of the
reaction occurs.
The other way to speed up a reaction is to reduce the barrier, as shown by the
dashed line in the diagram. When less energy is needed for the reaction, more
molecules will possess enough energy to get over the barrier. Reducing the
barrier is the job of catalysts. They alter the situation to reduce the barrier to
reaction. Enzymes are a subset of catalysts; they work on biological reactions.
About 4000 reactions are known to involve enzymes, including most of the
reactions that occur in the human body and several reactions in bread dough,
described next.

Enzymes
catalyze three main reactions in bread-making: breaking starch into maltose, a
complex sugar; breaking complex sugars into simple sugars; and breaking
protein chains. The breakages could happen without enzymes, but the energy
barrier is so large that it is very unlikely. Essentially, the enzymes are
necessary for the reactions to occur.
It is easy to start seeing enzymes as little critters that come in, recognize the
site where they can work, and begin to chew on bonds or snap them in half.
While this is a convenient picture, it does a disservice to the marvels of
biology. Enzymes do not think or act, but still manage to arrive at the sites
where they are needed. Each enzyme has a very specific job to do and only
interacts with the appropriate molecules for which it is designed, ignoring all
others. Enzymes work efficiently and are not used up by the process; after the
reaction occurs, the original enzyme molecule is left intact and can proceed to
a new site.
If the enzyme does not think, how does it manage to perform its specific task?
The simplified picture presented in general chemistry textbooks is called the
“lock and key model.” An enzyme has a specific shape that fits together with
the substrate, the molecule on which it will be working. The enzyme bonds to
the substrate with a weaker chemical bond, a hydrogen bond or hydrophobic
bond, for example. It alters the substrate in a way that makes reaction
favorable. Once reaction occurs, the enzyme releases the products and moves
on.
For example, the substrate sucrose is a complex sugar that can react with a
water molecule to form two simple sugar molecules, glucose and fructose.
 There is an
energy barrier to the reaction because it takes a lot of energy to break the
middle bond of the sucrose.
The enzyme sucrase fits together with the sucrose (below). In order to bond to
the enzyme, the sucrose must stretch. This stretching weakens the sucrose’s
middle bond, which becomes susceptible to attack by water molecules. The
energy barrier has been lowered. When a water molecule comes along, the
middle bond easily breaks and reacts with the water molecule. The enzyme is
now holding the product molecules, which it releases. Sucrose has been
broken into glucose and fructose.
Another example emphasizes the bonding nature of the enzymes; they are not
simply fitting into substrates like puzzle pieces, snapping into place. Bonds
must form. Once bonded, the active site of the enzyme is positioned near the
reaction site of the substrate, which it alters to reduce the energy barrier.

In this example, the substrate is a protein. Proteins are chains of amino acids
linked by peptide bonds. When a peptide bond forms, a water molecule is
released.

A water
molecule can come back and break a peptide bond, but it usually does not
have enough energy.
The enzyme carboxypeptidase catalyzes the breaking of the last peptide bond
in the protein chain, releasing the end amino acid. Carboxypeptidase contains
a zinc atom with a positive charge. This zinc atom bonds with the protein near
the last peptide bond, pulling the electrons of the bond away from it and, thus,
weakening it (below). The enzyme also has a pocket area composed of
hydrophobic atoms; if the terminal amino acid has a hydrophobic group on it,
the group is attracted to this pocket and held by it. In addition,
carboxypeptidase can form hydrogen bonds with the terminal amino acid,
further securing it in place.
When a water molecule encounters the weakened peptide bond, it likely now
has enough energy to break it, recombining with the broken ends to reform
the loose amino acid. The various bonds holding the enzyme to the protein
substrate are weakened, and the enzyme is released.
The first enzyme to take action in bread dough is amylase. Amylase acts on
starch (either amylose or amylopectin), breaking the starch chain between
adjacent sugar rings. There are two kinds of amylase: α-amylase (alpha-
amylase) randomly breaks the chain into smaller pieces while β-amylase
(beta-amylase) breaks maltose units off the end of the chain.

Amylase is found in flour. Wheat kernels contain amylase because they need
to break starch down into sugar to use for energy when the kernels germinate.
The amount of amylase varies with the weather and harvesting conditions of
the wheat, so mills generally test for it and add extra or blend flours to get an
appropriate amount.

Amylases are mobilized when water is added to the flour. This is one reason
why doughs with a higher hydration often ferment faster—the amylases (and
other enzymes) can move about more effectively. To reach the starch
molecules, amylases must penetrate the starch’s granules; thus, most of the
action in bread dough happens at broken granules, where the starch is
available for reaction. Fortunately, a percentage of starch granules are
damaged during milling and accessible by the amylases.

An amylase is a big molecule, with hundreds of amino acids linked together.

Many different groups contribute to the bonding between the amylase and the
starch substrate. In addition, there are several different amylase molecules,
and each functions differently. The examples of enzyme action presented
above give the general idea.
Because of amylase, some of the starch in bread dough is broken into maltose,
a double-ring sugar composed of two glucose molecules; but fermentation
reactions require single glucose rings. Simple sugars like glucose also provide
flavor to the bread and participate in browning reactions that occur at the
crust during baking.
Fortunately, the yeast used in bread-making contains the enzyme maltase,
which breaks maltose into glucose. When the yeast cell encounters a maltose
molecule, it absorbs it. Maltase then bonds to the maltose and breaks it in two.
Yeast cells also contain invertase, another enzyme that can break sucrose, like
the sucrase described above. This enzyme works on the small percentage of
sucrose found in the flour. These two enzymes are responsible for producing
much of the glucose needed by the yeast for fermentation.
The other major enzyme at work in bread dough is protease. Protease acts on
protein chains, breaking the peptide bonds between amino acids.
Carboxypeptidase, described above, is an example of a protease. There are
hundreds of proteases, but only a few are found in bread dough, where they
chop the gluten into pieces. Proteases occur naturally in flour, yeast cells, and
malt. Their levels are measured at the mill and adjusted in the same way that
amylase levels are adjusted.

Proteases in bread dough have been the subject of scientific research for the
past hundred years. There has been much debate about their importance. In
the early years, scientists were trying to prove their existence and measure
relative activity in different brands of flour. They amplified the protease
activity by adding non-gluten substrates to the mix. These substrates were
ones that protease readily attacks. Eventually someone thought to look at
protease activity in normal bread dough and found very little activity.

It seems, however, that this very small activity might be just what is needed in
bread dough. Too much protease activity would break up the gluten,
destroying the network that forms during kneading. A little bit, however,
softens the dough and makes it more workable. If the dough is allowed to
autolyse (i.e., rest) or if preferments are used, proteases have time to work
before kneading, making the dough easier to knead. (I wonder if this is the
origin of the word “autolyse,” from “autolysis,” which means “self-breaking”
and could refer to the protein proteases at work on the protein chains.)

In addition to affecting the dough’s consistency, proteases affect its flavor.

Proteases result in single amino acids when they break the last peptide bond
of the protein chain. These amino acids can participate in the flavor and
browning reactions that occur at the crust during baking.

So now, my simplified diagram of the chemical reactions in bread dough looks

more like this:

This diagram includes the presence of enzymes. Without enzymes, bread-

making would not be possible. Then again, neither would we.
Introduction
Have you ever wondered how all the food that you eat gets digested? It is not
only the acid in your stomach that breaks down your food—many little
molecules in your body, called enzymes, help with that too. Enzymes are
special types of proteins that speed up chemical reactions, such as the
digestion of food in your stomach. In fact, there are thousands of different
enzymes in your body that work around-the-clock to keep you healthy and
active. In this science activity you will investigate one of these enzymes, called
catalase, to find out how it helps to protect your body from damage.
Background
Enzymes are essential for our survival. These proteins, made by our cells, help
transform chemicals in our body, functioning as a catalyst. A catalyst gets
reactions started and makes them happen faster, by increasing the rate of a
reaction that otherwise might not happen at all, or would take too long to
sustain life. However, a catalyst does not take part in the reaction itself—so
how does this work? Each chemical reaction needs a minimum amount of
energy to make it happen. This energy is called the activation energy. The
lower the activation energy of a reaction, the faster it takes place. If the
activation energy is too high, the reaction does not occur.
Enzymes have the ability to lower the activation energy of a chemical reaction
by interacting with its reactants (the chemicals doing the reacting). Each
enzyme has an active site, which is where the reaction takes place. These sites
are like special pockets that are able to bind a chemical molecule. The
compounds or molecules the enzyme reacts with are called their substrates.
The enzyme pocket has a special shape so that only one specific substrate is
able to bind to it, just like only one key fits into a specific lock. Once the
molecule is bound to the enzyme, the chemical reaction takes place. Then, the
reaction products are released from the pocket, and the enzyme is ready to
start all over again with another substrate molecule.
Catalase is a very common enzyme that is present in almost all organisms that
are exposed to oxygen. The purpose of catalase in living cells is to protect them
from oxidative damage, which can occur when cells or other molecules in the
body come into contact with oxidative compounds. This damage is a natural
result of reactions happening inside your cells. The reactions can include by-
products such as hydrogen peroxide, which can be harmful to the body, just as
how a by-product of a nice bonfire can be unwanted smoke that makes you
cough or stings your eyes. To prevent such damage, the catalase enzyme helps
getting rid of these compounds by breaking up hydrogen peroxide (H2O2) into
harmless water and oxygen. Do you want to see the catalyze enzyme in action?
In this activity you will disarm hydrogen peroxide with the help of catalase
from yeast.
Materials
 Safety goggles or protective glasses
 Five teaspoons of dish soap
 One package of dry yeast
 Hydrogen peroxide, 3 percent (at least 100 mL)
 Three tablespoons
 One teaspoon
 Five 16-ounce disposable plastic cups
 Tap water
 Measuring cup
 Permanent marker
 Paper towel
 Workspace that can get wet (and won't be damaged by any spilled hydrogen
peroxide or food-colored water)
 Food coloring (optional)
Preparation
 Take one cup and dissolve the dry yeast in about one-half cup of warm tap
water. The water shouldn't be too hot but close to body temperature (37
Celsius). Let the dissolved yeast rest for at least five minutes.
 Use the permanent marker to label the remaining four cups from one to four.
 To all the labeled cups, add one teaspoon of dish soap.
 To cup one no further additions are made at this point.
 Before using the hydrogen peroxide, put on your safety goggles to protect your
eyes. In case you spill hydrogen peroxide, clean it up with a wet paper towel. If
you get it on your skin, make sure to rinse the affected area with plenty of
water.
 To cup two, add one tablespoon of 3 percent hydrogen peroxide solution. Use
a fresh spoon for the hydrogen peroxide.
 To cup three, add two tablespoons of the hydrogen peroxide.
 To cup four, add three tablespoons of the hydrogen peroxide.
 Optionally, you can add a drop of food color to each of the labeled cups. (You
can choose a different color for each one for easy identification)
Procedure
 Take cup number one and place it in front of you on the work area. With a
fresh tablespoon, add one tablespoon of the dissolved yeast solution to the cup
and swirl it slightly. What happens after you add the yeast? Do you see a
reaction happening?
 Place cup number two in front of you and again add one tablespoon of yeast
solution to the cup. Once you add the enzyme, does the catalase react with
the hydrogen peroxide? Can you see the reaction products being formed?
 Add one tablespoon of yeast solution to cup number three. Do you see the
same reaction taking place? Is the result different or the same compared to
cup number two?
 Finally, add one tablespoon of yeast solution to cup number four. Do you see
more or less reaction products compared to your previous results? Can you
explain the difference?
 Place all four cups next to each other in front of you and observe your
results. Did the enzymatic reaction take place in all of the cups or was there
an exception? How do the results in each cup look different? Why do you
think this is the case?
 Now, take cup number one and add one additional tablespoon of 3 percent
hydrogen peroxide to the cup. Swirl the cup slightly to mix the solution. What
happens now? Looking at all your results, what do you think is the limiting
factor for the catalase reaction in your cups?
 Extra: Repeat this activity, but this time do not add dish soap to all of the
reactions. What is different once you remove the dish soap? Do you still see
foam formation?
 Extra: So far you have observed the effect of substrate (H2O2) concentration
on the catalase reaction. What happens if you keep the substrate
concentration constant but change the concentration of the enzyme? Try
adding different amounts of yeast solution to three tablespoons of hydrogen
peroxide, starting with one teaspoon. Do you observe any differences, or does
the concentration of catalase not matter in your reaction?
 Extra: What happens if the environmental conditions for the enzyme are
changed? Repeat the catalase reaction but this time vary conditions such as
the pH by adding vinegar (an acid) or baking soda (a base), or change the
reaction temperature by heating the solution in the microwave. Can you
identify which conditions are optimal for the catalase reaction? Are there
any conditions that eliminate the catalase activity?
 Extra: Can you find other sources of catalase enzyme that you could use in
this activity? Research what other organisms, plants or cells contain catalase
and try using these for your reaction. Do they work as well as yeast?
Observations and results
You probably saw lots of bubbles and foam in this activity. What made the
foam appear? When the enzyme catalase comes into contact with its substrate,
hydrogen peroxide, it starts breaking it down into water and oxygen. Oxygen is
a gas and therefore wants to escape the liquid. However, the dish soap that
you added to all your solutions is able to trap the gas bubbles, which results in
the formation of a stable foam. As long as there is enzyme and hydrogen
peroxide present in the solution, the reaction continues and foam is produced.
Once one of both compounds is depleted, the product formation stops. If you
do not add dish soap to the reaction, you will see bubbles generated but no
stable foam formation.
If there is no hydrogen peroxide present, the catalase cannot function, which
is why in cup one you shouldn't have seen any bubble or foam production.
Only when hydrogen peroxide is available, the catalase reaction can take place
as you probably observed in the other cups. In fact, the catalase reaction is
dependent on the substrate concentration. If you have an excess of enzyme but
not enough substrate, the reaction will be limited by the substrate availability.
Once you add more hydrogen peroxide to the solution, the reaction rate will
increase as more substrate molecules can collide with the enzyme, forming
more product. The result is an increasing amount of foam produced in your
cup as you increase the amount of H2O2 in your reaction. You should have seen
more foam being produced once you added another tablespoon of hydrogen
peroxide to cup one, which should have resulted in a similar amount of foam
as in cup two. However, at some point you will reach a substrate concentration
at which the enzyme gets saturated and becomes the limiting factor. In this
case you have to add more enzyme to speed up the reaction again.

Many other factors affect the activity of enzymes as well. Most enzymes only
function under optimal environmental conditions. If the pH or temperature
deviates from these conditions too much, the enzyme reaction slows down
significantly or does not work at all. You might have noticed that when doing
the extra steps in the procedure.
Cleanup
Pour all the solutions into the sink and clean all the spoons with warm water
and dish soap. Wipe your work area with a wet paper towel and wash your
hands with water and soap.
(https://www.scientificamerican.com/article/exploring-enzymes/)

Hydrogen Peroxide Breakdown in Liver vs. Potato

What gas was produced by the breakdown of hydrogen peroxide?

Oxygen gas was produced
Describe the test that was performed in order to identify the gas.
A glowing splint of a match was lit, blown out then inserted into the test tube. The match
relighting in the test tube indicates oxygen gas is present.
Can hydrogen peroxide be broken down by catalyst other than those found in a living
system?
Hydrogen peroxide can be broken down by manganese dioxide because it has catalytic
properties. It is unstable which makes it very reactive. It even breaks down in the presence
of light. It increases the rate of reaction without being changed. Sand however is not able to
break it down because it contains no catalytic properties.
Explain how temperature affected the enzyme’s function
Increasing the temperature increased the rate of reaction. There is a higher energy when
heated. The enzyme was able to catalyze the reaction more quickly. This is only until the
point until denaturation. At 40 degrees, the enzyme would experience denaturation causing
the rate of reaction to drop. The enzyme would be damaged and not be able to perform the
same way.
How did particle size affect the rate of reaction?
Smaller size of particles increased the rate of reaction because smaller particles consume
less energy than larger ones to break down molecules, therefore the reaction would happen
faster. Larger particles decreased the rate of reaction because they require more energy to
break down.
Explain why there is a difference in the rates of reaction between the liver and the
potato
Liver contains more of the enzyme catalase, which breaks down hydrogen peroxide. Liver
contains more because it detoxifies substances in the body. A larger amount of catalase
lowers the activation energy, therefore speeds up the rate of reaction. The potato contains
less of the enzyme catalase, therefore requires more activation energy, slowing down the
rate of reaction.
Show the fully labeled balanced chemical equation for the decomposition of
hydrogen peroxide
2 H2O (aq) –(catalase)—> 2 H2O (l) + O2 (g)
hydrogen peroxide enzyme water oxygen gas
Why is it possible to use dead cells to study the function of this enzyme?
Although the cells are dead, catalase still remains active. It remains active in certain
temperatures up until the point of denaturation which occurs above 40 degrees. The
organism which contained the cells is gone but the cells are still present and active in
certain conditions.
(https://schoolworkhelper.net/hydrogen-peroxide-breakdown-
liver-vs-potato/)

Introduction
Your liver is important for cleaning up any potentially dangerous substances
you consume. But how does it do it?—With a little help from some complex
chemistry. Within your liver, as within every tissue in the body, many
chemical reactions occur. Often these reactions require "help" to happen at a
faster speed, and this can be supplied by enzymes—tiny types of proteins.
The liver uses specialized enzymes to help it break down toxic substances and
make them safer for the body to process. But an enzyme, just like the chemical
reactions it modifies, needs certain conditions to do its work. So, some
environments can make a liver enzyme effective, whereas others can prevent it
from working at all.
Background
A chemical reaction occurs when compounds come together and their
molecules interact to form new compounds. Sometimes these reactions
happen by themselves, are usually very fast and spontaneous, and give off
energy. Other chemical reactions need energy, without which they would
proceed very slowly or not at all. Enzymes can help speed up these types of
chemical reactions.

Enzymes are large proteins that speed up the rate of a chemical reaction by
acting as a catalyst. A catalyst provides the necessary environment for the
reaction to occur, thereby quickening it. Certain catalysts work for certain
kinds of reactions; in other words, each enzyme has a particular type of
reaction that it can activate. Enzymes can be very fussy and sometimes need to
be in certain environments or conditions to work well—or at all. Some
enzymes can even be damaged, such as when exposed to too much heat. A
damaged enzyme may no longer work to catalyze a chemical reaction.
Catalase is an enzyme in the liver that breaks down harmful hydrogen
peroxide into oxygen and water. When this reaction occurs, oxygen gas
bubbles escape and create foam.

Materials
• Raw liver (fresh or frozen, thawed; one quarter pound)
• Knife
• Cutting board
• Blender
• Water
• Refrigerator
• Medicine dropper
• Large plate
• Hydrogen peroxide (new or recently purchased bottle works best)
• Measuring teaspoon
• Two bowls
• Vinegar
• Baking soda
• Microwave-safe bowl (with a cover)
• Microwave oven
Preparation
• Completely disinfect any surface that the raw liver touches during this
activity.
• On the cutting board, carefully cut the liver into little, cube-shaped pieces,
about one to two centimeters long. Be careful using the sharp knife. (An adult
may need to help with this.)
• Place the liver cubes into a blender and add an equal volume of water.
Blend on high speed, pulsing when necessary, until the liver is smooth and no
chunks are present. Be careful of the sharp blades in the blender.
• Keep the blended liver in the refrigerator.

Procedure
• Put one drop of the blended liver on the large plate. To the blended liver
drop, add one drop of hydrogen peroxide. You should see a lot of
bubbles! What do you think the bubbles are made of? This shows that the
liver enzyme catalase is working to start the chemical reaction that breaks
down the hydrogen peroxide that would be harmful to the body into less
dangerous compounds.
• To test the effect of an acid on the liver enzyme, put one teaspoon of the
blended liver in a bowl and mix it well with one teaspoon vinegar. What is the
color and consistency of this mixture? Put one drop of the mixture on a clean
part of the large plate and add one drop of hydrogen peroxide to it. Compared
with the untreated blended liver, did more, less or about the same amount of
bubbles form? Did they form more slowly, more quickly or at about the same
rate?
• To test the effect of a base, put one teaspoon of the blended liver in a bowl
and mix it with one teaspoon baking soda. What is the color and consistency
of this mixture? Put one drop of the mixture on a clean part of the large plate
and add one drop of hydrogen peroxide to it. Did more, less or about the same
amount of bubbles form? Did they form more slowly, more quickly or at
about the same rate?
• To test the effect of heat, put one teaspoon of the blended liver into a
microwave-safe bowl. Cover the bowl and microwave it on high for 20
seconds. How does the blended liver look after heating? Remove a drop-size
amount of the heated liver and put it on a clean part of the large plate. Add
one drop of hydrogen peroxide to it. Did more, less or about the same amount
of bubbles form? Did they form more slowly, more quickly or at about the
same rate?
• Based on your observations, under which condition(s) does it look like the
enzyme works best? Which condition(s) makes it work the worst? Why do
you think this is so?
• Extra: Try experimenting with other conditions. For example, try freezing
some blended liver or mixing it with salt and then test the enzyme's activity.
Or you could try adding more than one teaspoon of vinegar or baking soda and
then test the enzyme. Under which conditions does the enzyme work well,
and under which ones does it work poorly?
• Extra: You could try this activity again using another enzyme, called
bromelain, which digests proteins and can be extracted from pineapples. One
protein that is fun to digest using bromelain is gelatin, which is found in many
puddings and gelatinous desserts. How do different conditions affect the
ability of bromelain to digest proteins?

Observations and results

When exposed to hydrogen peroxide, did the blended liver bubble less when
mixed with either the vinegar or baking soda compared with when it was
untreated? Did it bubble even less after it was microwaved?

An enzyme needs certain conditions to work, and the ideal environment can
be a hint as to where the enzyme normally works in the body. And because
different body tissues have distinct environments—acidic or warm—each
enzyme is tuned to work best under specific conditions.

Different tissues in the body have different pHs (pH is a measure of how basic
or acidic a solution is). The liver maintains a neutral pH (about pH 7), which is
easiest for its enzymes, such as catalase, to work in. Consequently, when
exposed to hydrogen peroxide the liver should have produced more bubbles
(oxygen gas), and at a faster rate, when it was untreated than when exposed to
vinegar or baking soda. (It may have bubbled more when treated with baking
soda, compared with vinegar, because it might have been better able to return
the pH to around 7.)

Similarly, enzymes in the liver are also used to functioning at body

temperature (37 degrees Celsius), so microwaving the blended liver to a
temperature hotter than that should have damaged the catalase enzyme and
clearly decreased the amount of bubbles when it was exposed to hydrogen
peroxide.
Cleanup
Safely dispose of any raw liver meat used in this activity by putting it in the
trash when you are done. Completely disinfect any surfaces that the raw liver
 meat touched during this activity, and be sure to thoroughly wash your hands
with soap and warm water.
(https://www.scientificamerican.com/article/bring-science-home-liver-helping-enzymes/)

How do living cells interact with the environment around them? All living things
possess catalysts, or substances within them that speed up chemical reactions and
processes. Enzymes are molecules that enable the chemical reactions that occur in all
living things on earth. In this catalase and hydrogen peroxide experiment, we will
discover how enzymes act as catalysts by causing chemical reactions to occur more
quickly within living things. Using a potato and hydrogen peroxide, we can observe how
enzymes like catalase work to perform decomposition, or the breaking down, of other
substances. Catalase works to speed up the decomposition of hydrogen peroxide into
oxygen and water. We will also test how this process is affected by changes in the
temperature of the potato. Is the process faster or slower when compared to the control
experiment conducted at room temperature?
Problem
What happens when a potato is combined with hydrogen peroxide?
Materials

 1 Potato
 Hydrogen peroxide
 Small glass beaker or cup

Procedure

1. Divide the potato into three roughly equal sections.

2. Keep one section raw and at room temperature.
3. Place another section in the freezer for at least 30 minutes.
4. Boil the last section for at least 5 minutes.
5. Chop and mash a small sample (about a tablespoon) of the room temperature potato
and place into beaker or cup.
6. Pour enough hydrogen peroxide into the cup so that potato is submerged and observe.
7. Repeat steps 5 & 6 with the boiled and frozen potato sections.

Observations & Results

Watch each of the potato/hydrogen peroxide mixtures and record what happens. The
bubbling reaction you see is the metabolic process of decomposition, described
earlier. This reaction is caused by catalase, an enzyme within the potato. You are
observing catalase breaking hydrogen peroxide into oxygen and water. Which potato
sample decomposed the most hydrogen peroxide? Which one reacted the least?
Why?
You should have noticed that the boiled potato produced little to no bubbles. This is
because the heat degraded the catalase enzyme, making it incapable of processing the
hydrogen peroxide. The frozen potato should have produced fewer bubbles than the
room temperature sample because the cold temperature slowed the catalase enzyme’s
ability to decompose the hydrogen peroxide. The room temperature potato produced
the most bubbles because catalase works best at a room temperature.
Conclusions
Catalase acts as the catalyzing enzyme in the decomposition of hydrogen peroxide.
Nearly all living things possess catalase, including us! This enzyme, like many others,
aids in the decomposition of one substance into another. Catalase decomposes, or
breaks down, hydrogen peroxide into water and oxygen.
Want to take a closer look? Go further in this experiment by looking at a very small
sample of potato combined with hydrogen peroxide under a microscope!
Author: Justine Rembac
(https://www.education.com/science-fair/article/activator/)

What are enzymes?

(http://brilliantbiologystudent.weebly.com/enzymes.html)
Structure of Chymotrypsin, a protease enzyme which is active in
the duodenum.
Enzymes are biological catalysts. A catalyst is a substance
which speeds the rate of reaction but remains unchanged in the
reaction. Catalysts reduce the activation energy needed for a
reaction. Enzymes are proteins and occur naturally in living
biological systems, acting in many metabolic pathways.

The activity of enzymes are affected by pH, temperature, enzyme

concentration and substrate concentration. Enzymes have optimum pH and
optimum temperatures, at which they experience maximal activity.

Enzymes are highly specific, acting upon a single substrate or group of related
substrates. Enzymes have an active site - a small portion of the molecule which is
complementary in shape to a portion of the substrate. The substrate binds to the active
site of the enzyme forming the enzyme-substrate complex. Strain is induced in the
bonds causes them to cleave and the the products leave the active site, leaving it
available for further reactions. The reaction of enzymes are reversible.
Learning Objectives
  Explain the properties and functions of enzymes
 Explain how various factors - pH, temperature, substrate concentration, enzyme
concentration - can affect the rate of enzyme-catalyzed activity
 Explain the action of inhibitors - competitive or non-competitive, reversible or
irreversible.

Enzyme Experiments

 To investigate the effect of substrate concentration on enzyme activity

To investigate the effect of enzyme concentration on enzyme activity
To investigate the effect of temperature on enzyme activity
To investigate the effect of pH on enzyme activity

To investigate the effect of substrate concentration on enzyme activity

 To investigate the effect of different pH values on enzyme activity

 To investigate the effect of temperature on enzyme activity

Enzyme and substrate pairs commonly used in the for biology experiments are:

 Catalase (from raw potato tuber) & hydrogen peroxide

 Amylase (commercial preparation) & starch
 Sucrase - invertase is the most common sucrase ( commercial preparation)
& sucrose.

Factors affecting Enzyme activity

(http://www.nuffieldfoundation.org/practical-biology/factors-affecting-enzyme-activity)
Enzymes are sophisticated catalysts for biological processes. These practicals (and the
practicals at intermediate level) give you opportunities to explore how enzyme activity
changes in different conditions.
Enzyme experiments often provide real ‘messy’ data, because their activity can change
dramatically from one lesson to the next.
 Microscale investigations of catalase activity in plant extracts
Adsorb microscale quantities of plant extract onto filter paper discs and assess
catalase activity by comparing times taken for the discs to surface in hydrogen
peroxide solution.
 Investigating an enzyme-controlled reaction: catalase and hydrogen peroxide
concentration
Use catalase in pureed potato and investigate the effect of changing hydrogen
peroxide concentration on rate of production of oxygen.
 Investigating effect of temperature on the activity of lipase
A simple protocol that measures the effect of changing temperature on the time taken
for lipase to break down the fat in milk to form fatty acids (and glycerol).
 Investigating the effect of pH on amylase activity
Measure the time taken for amylase to completely break down a sample of starch,
using buffers for different pHs.
 Investigating effect of concentration on the activity of trypsin
Measure the time taken for different concentrations of trypsin to digest the gelatine
coating on exposed photographic film and release the blackened silver halides.