Name: Stainley Yap Lik Hong

Class:DCB1(A2)

Lecture:

Course:AACB1253 Bioenergetics
Introduction

Enzymes are macromolecular catalyst which provides an alternative path way to accelerate the process
of a chemical reaction which required lower activation energy. Enzyme may convert substrates to another
product which usually happen in a current existing cell. This reaction will be done through the temporary
binding of the enzymes to one or more reactants. Enzymes are widely knew as a proteins and currently
research shown that not all enzymes are proteins as some are ribonucleoprotein enzymes where catalytic
activity is in the RNA part rather than in the protein part. (Enzymes, 2016, Enzymes, Para. 1 The Editors
of Encyclopedia Britannica, 2016 Westfall, 2013). Furthermore, Alajar(2009) also stated that, Some
enzymes are composed chiefly of proteins while most [are made up of] protein and non-proteinportions
which can either be an organic molecule or a metal ion" known as a cofactor (Enzymes, 2015, Function
and structure, Para. 4).Almost all metabolism process need enzyme to accelerate their chemical process
to sustain life. Enzymes are currently known as catalyst almost 5000 types of chemical reaction. Usually
most of the persons confused about different between of enzymes and catalyst, but there are little
different between both of them.There are mainly two types of catalyst group which are competitive and
non comepetitive inhibitor.Competitive inhibitor function by binding reversibly to the active site of
the enzyme however non competitive inhibitor function by bind to the enzyme whether or not
the substrate has already been bound, but if it has a higher affinity for binding the enzyme in one
state or the other.Catalyst is an enzyme used by the liver to decompose the poisonous hydrogen peroxide
(H2O2) into oxygen (O2) and water (H2O). This experiment seeks to prove that catalyst is the cause of this
reaction and that oxygen and water are the byproducts, and also to explore the effects of increased
temperature on catalyst activity, and enzyme activity in general. The main different is that enzymes are
largely organic in nature and are bio-catalysts, while non-enzymatic catalysts can be inorganic compounds
such as Iron, vanadium(V) oxide, platinum , rhodium and so on.A catalyst is a chemical that increases or
decrease the rate of a chemical reaction remain unchanged after the reaction. In fact that they aren't
changed by participating in a reaction distinguishes catalysts from substrates, which are the reactants on
which catalysts work but enzymes catalyze biochemical reactions. So, enzymes are proteins that increase
rate of chemical reactions converting substrate into product. Catalyst also not specific and therefore end
up producing residues with errors but enzyme are highly specific producing large amount of residues. The
reaction of a catalyst required a high temperature and pressure but enzyme required a good condition
and environment which provide an optimal pH, temperature, salt condition and so on. Then the reaction
rates of a catalyst typical slower compare with enzyme.The enzyme activity can be controlled but the
activity of the catalysts can not be controlled.Therefore, all enzymes count as catalysts but not all
catalysts are enzymes.

The similar between enzymes and catalysts they are both remain unchanged chemically and
quantitatively at the end of a reaction and they can be recycle using.Then, they are both are
required in small quantity as compared to their substrates and do not change the equilibrium of
chemical reaction.After that,the reaction control by catalyst and enzyme are reversible and
They lower the activation energy needed to start the reaction. Enzyme and catalyst are both
also only enhance the rate of chemical reaction and do not initiate the reaction.Other than
that, They form short-lived complexes with the substrate molecules and does not change the
product after the reaction.

Every enzyme is specific for some reaction and enzyme are mainly possess great catalytic power.
Enzyme are also highly specific and show varying degree of specificities.Enzyme usually only bind
with one substrate.The reaction of an enzyme is highest while the enzyme is at the optimum
temperature also have an optimum pH range within which the enzymes function is at its
peak. The increase of concentration of the reactants, and substrate the rate of the reaction
increase until the enzyme will become saturated with the substrate.Otherwise, increase the
amount of enzyme, increase the rate of the reaction.Some of the.Inorganic substances known as
activators also increase the activity of the enzyme.

There are many factors will affect the efficiency of enzyme such as pH, temperature, salt concentration,
small molecules.Here are the examples how these factors affect the activity of enzyme.

pH=pH is a unit to measure acidity and basicity of a solution.pH is a measure of the hydrogen

Ion (H+) concentration. and therefore a good indicator of the hydroxide Ion (OH-)

concentration. It ranges from pH1 to pH14. Lower pH values mean higher H+ concentrations

and lower OH- concentration.Acid solutions have pH values below 7, and Basic solutions have

pH values above7, Deionised water is pH7, which is termed 'neutral'.The change of pH will
alter the enzyme shape and become denatured.However, different enzymes work best at
different pH values.The optimum pH of each enzyme is variable and depend on where it
normally works. For examples, intestinal enzymes such as trypsin has an optimum pH of about
7.5 . Enzymes in the stomach has an optimum pH of about 2.

Temperature-As the temperature increases, the rate of reaction also increased,but too high
temperature will denature enzyme.The activity of enzyme will increase dramatic increase with
temperature up to around 37 degree celsius or body temperature. Then as the temperature
continue increasing, the rate of reaction will fall rapidly as the heat energy cause denatures the
enzyme.

Salt concentration-Salt concentration could affect the activities of enzymes by high

concentration of salt cause change in salinity.Thus it will cause add or remove of cation and
anion and it also disrupt bond of enzyme by disrupt the attraction between charged amino
acids at the end the protein of enzyme will become denatured by affect the structure of
protein.

Aim

To test and analyze that how the pH affect the rate of reaction of enzyme catalyst

Materials

Fresh liver, potatoes, sand,10ml measuring cylinder, dilute hydrochloric acid solution, 100ml hydrogen
peroxide, boiling water, mortar and pestle, toothpick, 10ml measuring pipettes, dilute sodium hydroxide
acid, clean test tubes and vials

Method

1)5 mi of 10 V hydrogen solution which measured by student then poured into each eight labeled
vial by using measuring cylinder.
2) One piece of liver cubes was taken by using a toothpick ,then placed into boiling for 2-3
minutes.Student should remove and allow it to cool before started next step.
3)Student should take another cube of liver and another of potato, place into separate
mortals.Release the cell content by grinding each tissue with a separate pestle.
4)To the vials labeled A to F, following table was added:

Vial Solution Substance added

A 5ml Hydrogen peroxide Pinch of sand

B 5ml Hydrogen peroxide Cube of liver

C 5ml Hydrogen peroxide Cube of potato

D 5ml Hydrogen peroxide Ground liver

E 5ml Hydrogen peroxide Ground potato

F 5ml Hydrogen peroxide Cube of boiled liver

G 5ml Hydrogen peroxide+1ml Dilute hydrochloric acid solution Cube of liver

H 5ml Hydrogen peroxide+1ml Dilute sodium hydroxide solution Cube of liver

 5)Number of bubbles evolved from the hydrogen peroxide in the vials observed by students.Speed
of production oxygen in a table recorded by students by using scale 0 to 5, where 0 represents no
production and 5 represent the most rapid production.
6)1 ml of dilute hydrochloric acid solution to vial G and 1 ml of dlute sodium hydroxide solution to
vial H.Then, cubes of liver was added to each vials.The rate of oxygen production observed and
recorded by students.

Result

Vial Solution Substance Speed of Observation

added production oxygen
(scale)

A 5ml Hydrogen peroxide Pinch of sand 0 No bubbles

formed actually

B 5ml Hydrogen peroxide Cube of liver 4 Bubble formed

immediately and
went up fast after
combining

C 5ml Hydrogen peroxide Cube of potato 2 Few bubbles

formed

D 5ml Hydrogen peroxide Ground liver 5 Bubbles formed

immediately
when reactants
touched and
overflowed

E 5ml Hydrogen peroxide Ground potato 3 Bubble formed

immediately but
didn't shoots
straight up

F 5ml Hydrogen peroxide Cube of boiled 1 Least bubble

liver formed, enzyme
could have
denatured
 G 5ml Hydrogen peroxide+1ml Dilute Cube of liver 3 Bubble formed
hydrochloric acid solution immediately but
didn't shoots
straight up
reactant

H 5ml Hydrogen peroxide+1ml Dilute Cube of liver 4 Bubble formed

sodium hydroxide solution immediately and
went up fast after
combining
reactant

Discussion and questions

1)What can you conclude from the results from tubes B and C?
Reaction of enzyme in liver in tube B faster than reaction of enzyme in potato in tube C.This is
because liver contain more enzyme catalyst compare with potato which breaking down hydrogen
peroxide become oxygen. Liver contain more because of it detoxified the body. A larger amount of
catalase lowers the activation energy, therefore speeds up the rate of reaction. The potato contains
less enzyme catalase, therefore requires more activation energy, slowing down the rate of reaction.

2)Comment on the results of the pairs of tubes A and E

Reaction of pinch of sand is nil however reaction of ground potato is happened.This is because the
sand does not contain any enzyme to break down hydrogen peroxide become oxygen however the
enzyme in cube potato in tube E able to break down hydrogen peroxide become oxygen because of
enzyme function well in that condition.

3)Comment on the results between tube B and F

Reaction of enzyme in liver in tube B formed bubbles immediately because of the enzyme is still able
to work however reaction of the enzyme in boiled liver of tube F formed least bubble because of
most of the enzyme denatured after undergo high temperature make them unable to break down
hydrogen peroxide immediately.

4)Compare the results from tubes B and G and tubes B and H

Reaction of enzyme in liver in tube B is faster than reaction of enzyme in liver in tube G because of
enzyme in liver in tube B is on a good environment which on the optimal pH value for the enzyme
however the enzyme in liver in tube H is on the alkali environment which will cause the enzyme
unable to function correctly.
 5)Write a summary on properties of enzymes.
The enzyme function quickest while at the optimal pH, temperature, salt concentration and smaller
molecules. When enzyme undergo boiled, it still able to function but might be very slow.Other than
that, enzyme will function better with smaller molecules as the reaction ground liver faster than
reaction of cube liver.Sand does not contain any enzyme so that it was unable to undergo reaction.
Enzyme at the alkali condition will undergo faster reaction as the reaction of tube H faster than
G.Thus, the speed order arrangement of the reaction are from tube A, F, C, E, G, B ,H , to D.

Coclusion

In this lab, we show that how pH and temperature will effect enzymes. We learned about pH
and temperature will effect on the enzyme. Enzymes are mainly work best at certain pHs, if
they are in a pH that is too low or too high they start to denature and don't function as well. In
this experiment, when we added NaOH to the enzyme, the reaction took place at a slower rate.
This can lead to the conclusion that if we were to make it more basic, the reaction may not
occur at all. This goes the same with adding the acid. When we added a couple drops of HCl,
there were no significant amount of bubbles that formed. This can conclude that the acidic
level was too high for the enzyme, preventing it to carry out its function. Additionally
temperature has the same effect. When we put the enzyme in the cold water bath, reaction
occurred at a slow rate, producing a small amount of bubbles. Then when we put the liver into
25 degree Celsius, the reaction sped up compared to the control. Enzymes in a high
temperature environment, will have their bonds break because they will start to denature.

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