Beruflich Dokumente
Kultur Dokumente
20 and 21
ENZYMES AND FACTORS INFLUENCING ENZYME ACTION
ENZYMES
Comes from the Greek words en which means in, and zyme which means yeast.
An organic compound that acts as a catalyst for a biochemical reaction.
o Catalyst speeds up the rate of biochemical reaction
o Enzyme is the most efficient catalyst that can increase the rates of reaction
by 1020
GENERAL STRUCTURAL CLASSES:
1)
2)
SIMPLE ENZYME
Is an enzyme composed of protein (amino acid chains))
CONJUGATED ENZYME
It has protein and non-protein part
o Apoenzyme is a protein part of a conjugated enzyme
o Cofactor - in a non-protein part of the conjugated enzyme ; provides
additional chemically reactive functional groups besides those present in
the amino acid chain of apoenzyme.
Vitamins (COENZYME) and METAL IONS are biochemically
important cofactors
Combination of the apoenzyme and cofactor enzyme produces a
biochemically active enzyme.
Apoenzyme + cofactor = Holoenzyme
CLASSES OF ENZYMES:
1) OXIREDUCTASE
An enzyme that catalyzes an oxidation-reduction reaction.
Oxidation reaction is an oxidation that increases the number of C-O bonds and/or
decreases the number of C-H bonds
Reduction reaction is a reduction that decreases the number of C-O and/or
increases the number of C-H bonds
Example:
2) TRANSFERASES
Is an enzyme that catalyses the transfer of a functional group from one molecule to
another
TWO MAJOR TYPES
o Transaminases catalyze the transfer of an amino group from one molecule
to another.
o Kinases catalyzes the transfer of a phosphate group from adenosine
triphosphate (ATP) to give adenosine diphosphate (ADP) and a
phospholrylated product containing an additional phosphate group; major
role in the metabolic energy-production reaction.
Example:
3) HYDROLASES
An enzyme that catalyzes a hydrolysis reaction in which the addition of water to a
molecule to a bond causes the bond to break.
It is the central process of digestion.
o The carbohydrases - effects the breaking of glycosidic bonds in oligo- and
polysaccharides
o The Proteases- effect the breaking of the peptide linkages inprotein
o The Lipases effect the breaking of ester linkages in triacyglycerols
Example
5) ISOMERASES
Is an enzyme that catalyzes the isomerization (rearrangement of atoms) of a
substrate in a reaction, converting it into a molecule isomeric with itself.
Example:
6) LIGASES
Is an enzyme that catalyses the bonding together of two molecules into one with
the participation of ATP.
o ATP involves is involved as energy provider.
4) LYASES
An enzyme that catalyzes the addition of a group to a double bond or the removal of
a group to form a double bond in a manner that does not involve hydrolysis or
oxidation.
o Dehydratase removal of water from a double bond
o Hydratase effects the addition of the component of water to a double
bond
Example:
MODELS OF ENZYME ACTION
ENZYME ACTIVE SITE
o Crevicelike location in the enzyme that participates in the in the
interaction with substrate during the reaction
o It is small part of an enzymes structure that is actually involved in catalysis.
SUBSTRATE
o The reactant in an enzyme catalyzed reaction
ACTIVITY NO. 20
A. POTATO OXIDASE
- OXIDASE is an enzyme that causes or promote oxidation.
- Read the article on the enzymatic browning: discoloration of fruits and
vegetables.
C. CATALASE
- CATALASE is an enzyme found in most living cells that catalyzes the separation
of hydrogen peroxide into gaseous oxygen and water.
C. TEMPERATURE
3 testtubes with the same amount of substrate and enzyme were exposed to
different temperature. The result shows that the color changes becomes more
intense as the temperature increase, and indication that temperature affects
enzyme action, wherein increasing temperature increases the enzyme activity.
Note: that enzyme exhibit optimum activity at 37 degrees celcius, beyond cause
enzyme denaturation, below causes enzyme inactivation.
ACTIVITY NO. 21
A. SUBSTRATE CONCENTRATION
In this set up the catechol (substrate) is increasing in each of the testube with the
same concentration of enzyme, result of the experiment shows that the intensity
of color reaction becomes darker as the concentration substrate increase the rate of
enzyme action increases to its maximum.
Scanpix pp202-203
B. ENZYME CONCENTRATION
In this set up the enzyme extractl (catalyst) is increasing in each of the test tube, the
result of the experiment shows that the intensity of color reaction becomes darker.
This concludes that as the concentration enzyme increase the rate of enzyme action
increases to its maximum.
D. pH
4 test tubes with the same amount of substrate and enzyme was exposed to
different pH level, the result shows that at acidic pH the solution becomes cloudy,
increasing pH (5-light red), (7-red brown) and (9-yellow) The result indicates that
optimum activity of an enzyme is obtain at neutral pH, because low or high pH
causes the enzyme to be deactivated or denatured
pH is neutral. Saliva is the first catalyst in the process of digestion. Enzyme amylase
can work efficiently at neutral pH. (exception on stomach enzyme.)
C. COMPOSITION OF SALIVA
TEST
NAME
OF
THE RESULT OBSERVED
CONCLUSION/
NO
TEST/REAGENT ADDED
PRESENCE OF
1
Molisch test
Violet ring formation
CHO
2
Biuret test
Purple color
2 peptide
3
Benedicts test
Blue
No reducing sugar
4
Dil. Acetic acid
White ppt
mucin
5
Dil. HNO3+AgNO3
White ppt
Cl6
Dil. HCL+BaCl2
White ppt
SO4
7
Dil.
HNO3+ammonium Yellow ppt
PO4
molybdate
8
Dil.
Acetic
acid+ White ppt
Ca++
(NH4)2C2)4
9
Dil.H2SO4 + KI+starch
Darck blue
NO2
10
FeCl3+Dil HCL
Yellow orange
Thiocyanate (SCN)
Post Lab Questions:
1) Same sample of saliva should be used to prevent variation of or results since the
composition varies depending on the physical state and genetic constituents of
the donor.
2) Factors responsible for the variation of the composition of saliva are as follows:
a. Mechanical factor- mastication of bland substance
b. Chemical factor action of sugars, acids and salts in the taste buds
c. Exposure to air- saliva lose CO2 and pH rises rapidly
d. Relative secretion of different glands, the release of more mucin, contain
less ptyalin.
3) Equation
a. AgNO3+Cl
AgCl ( white ppt)
b. Ba + SO4
BaSO4 (White ppt)
c. Ca + Ca2O4
CaC2O4
d. 2KI+2NO2
2KNO2+2I
e. FeCl3+3K
Fe(SCN)3+3KCL
4) Glycoprotein present in the saliva is the mucin, functions: a. Lubricant of the
mouth, B. Responsible for the viscosity of the saliva C. Protects lining of mouth
against drying.
A.
ACTIVITY NO. 23
PTYALIN
Determination of achromatic point
Iodine is the test for the presence of starch that gives dark blue color indicating a
positive result.
Benedicts reagent is a test for the presence of sugar, which give brick red ppt as a
positive result.
Beaker exposed to 40 degrees Celsius reached the achromatic point first (brown
color/no reaction) this means that the starch already dissociated and broken down.
Ptaylin an enzyme found in the saliva, having the property of converting starch into
dextrin and maltose thus aiding in the process of digestion.
B. TEMPERATURE
The test was done by exposing the ptyalin into four different temperature and the
result shows that test tube no 3 (40 degrees Celsius) reached the achromatic point
first. This indicates that ptyalin can work efficiently at optimum temperature which
is 37 degrees Celsius. Above the optimum temperature, its activity is hastened but is
Optimum pH 5.6-6.5, when the pH is lower than 4, its action is inhibited (Cabatit);
based on results of pH, below or above the optimum pH, activity ceases.
III. PRESENCE OF OTHER SUBSTANCES
1. Normal acidity of stomach contents is 1.6 -1.8 Stomach- columnar epithelium tining with
Halogens increases enzymatic activity but not in combination with heavy metals and
organic compounds
Chlorides, bromides and nitrates increases the activity.
ANSWERS TO QUESTIONS:
gastric pits containing gastric glands that produce gastric juice. Pepsin present digests
tissues of meat.
>
Answers to Questions:
protective mucus.
2. Lactic acid is secreted by gastric mucosa. Stagrration of gastric contents leads to bacterial
action and formation. Fermentation of CHO by lactic acid forming bacteria (Boas Oppler
Baciltus). Lactic acid in gastric juice:
a) bacterial fermentation
b) gastric carcincma
fermentation of
Abnormal:
Macroscopic Examination
l. Color
Normal: lighVdark browndue to stercobilin
Abnormal:
IV. Blood
- most common caused by ulcers, esophageal
- The higher the origin of biood in Gl tract, the
darker and more thoroughly mixed it will be with
feces.
Abnormal:
u,,v,vvvtvv Hl
hemorrhages.
prgments.
. Nature of diet
Example:
Rancid food- very sofl, watery/ ribbon like
High lipid- foamy and mushy
Thoroughly digested- hard and dry
Vegetarian- soft stool
Water intake and i-eabsorotion in colon.