INTRODUCTION
Proteins are compounds composed of the basic units; amino acids, containing carborn,
hydrogen and nitrogen (Debruyen, pinna & Whitney 2008). The amino acids bond together
through peptide linkages to form a protein basic chain and the chains interact through
different interactions, for example hydrogen bonding, forming complex shapes and
structures. Under different conditions that may lead to breakdown and change of protein
structure and their interactions, protein functionality also changes. Such conditions are
introduced to proteins through food processing and preparation.
Coagulation is one of the protein functionalities which results from denaturation.
Denaturation is the transformatiom of the folded protein structure into the unfolded state
through breakdown of interactions by the action of heat, enzymes and acid. Coagulation
results from the rearrangement of the denatured protein components and forming a
contineous sem-solid mass. Henceforth, the principle for protein cougulation experiment is
that proteim molecules get denatured by unwinding into the secondary or primary structures
and later clump together to form a gel when expeposed to heat or acid (McWilliams 2008).
The second experiment is on protein structure change through emzymatic browning. The
principle here is that foodstuffs as vegetables and fruits contain a phenolic compounds and
and an emzyme polyphenolase of which ender exposure to oxygen, the polyphenolase reacts
on the phenolic compound resulting imto the brown discoloration (McWilliams 2008).
It is therefore the objectives of the experiments to findout factors that cause protein
denaturation and the types of denaturation, and to illustrate the rate at which emzymatic
browning occurs and how to control it in fruits and vegetables.
MATERILAS AND METHODOLOGY
Eggs and milk were used for denaturation and coagulation experiments under heat, beating
and acid while potatos and apples were used for the emzymatic browning experiment at room
temparature, in boiled water, tap water and vinegar. Other materials include, bowls, whisk,
oven, cooker, testubes, litmas paper, pipette, pan, thermometer, knife, seive, crusher and a
chopping board.
On protein denaturation and coagulation, we experimented on the effect of exposure to air,
effect of PH and coagulation temperatures of proteins. On the exeperiment looking at the
effect of exposure to air, we separated an egg white from the egg yolk and beaten the egg
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white in a dry bowl with a whisk. After that the resulting product was divided into two
portions and placed the other at room temperature (exposed to air) and the other placed in an
oven at 90℃ and observed for 1 hour while recording the results. On the effect of PH, a
sample of milk was poured into a test tube and litmus paper was placed in. Using a pepette,
drops of vinegar were contineously added to the milk until coagulation begins to appear and
placed the litmas paper to record the resulting PH. On coagulation temperature of proteins,
the whole egg and the egg white were beaten separately and poured into different testtubes.
The test tubes were heated in simmerin water in a pan on a cooker. Using a thermometer, the
temperatures at which each of the samples start coagulation were recorded.
On emzymatic browning experiment, irish potatoes were peeled and cut into cubes and others
were crushed, and the apples were also cut into pieces. Then the samples were left under
different conditions as at room temperature, boiled water, tap water and in vinegar.
Addtionally, some potato pieces were first blanched and later treated as the rest of the
samples. The colour change was recorded over a 30 minutes period.
RESULTS AND DISCUSSION
Coagulation of egg and milk prrotein under different conditions
Table 1.0. Effects of exposure to air
Sample treatment Results
Beaten eggwhite and left at room Formation of Foam and is maintained and some
temperature liquid collects after sometime
Beaten eggwhite and left in an oven The foam is maintained and stable with some
hardening
Beating an egg causes agitation which denatures the protein structure and this to leads the egg
protein albumen to be spread over larger surface area so that air gets incoporated into the
bubbles created throu beating (McWllians 2008). This is why in the table, the foam is formed
due the incoporation of the air. McWilliams(2008) also argues that a foam is one of the
colloidal dispersions that involves the gases in the air being didpersed in in the liquid egg
white. During beating of the egg white to form the foam, some proteins are denatured and
then they agregate/coagulate adding rigidity and stability of the foam. Further denaturation
increases stability of the protein foam and this maybe through heating. This is why in the
table, the beaten eggwhite left exposed to air drains out some liquid because beating does not
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completely denature all the proteins such that they collect into their liquid native structures
over a specific perion of time. The beaten eggwhite left in an oven does not drain out some
liquid because the oven heat further dantures the protiens and their agregation briengs about
stability and rigidity. Hence beating does not give permanebt denaturation while heating
denaturation is irreversible.
Table 1.1. Effect of PH
Sample PH before PH after coagulation Changes
Milk 6.6 4.5 Formation of clumps or priciptates within
the milk
Here the principle is that adding an acid to milk will cause the casein protein to coagulate
(Brown 2011). In the table the ph of milk before adding acid vinegar is 6.6 determined by
litmus paper as it did slightly not change its blueish colour but after addind vinegar, the the
litmus paper turned pink to redish colour an indicator of increasing acidity. Some clumps also
formed after the addition of the vinegar. Again Brown(2011), says that “casein protein
preciptate when the normal 6.6 pH of milk drops below 4.6” (pg. 225). Thus, the pricipitates
of casein are as a result of coagulation. Vinegar has a Ph of 4 and adding it to the slightly
acidic milk will definately increase acidity (4.5ph) hence the litmus paper turns red (Earl &
Wilford 2001). This is why we see coagulation and is ofcourse irrevesible.
Table 1.2. Coagulation temperature of proteins
Sample Coagulation temperature
Beaten whole egg 68
Beaten eggwhite 62
The table above shows that beaten whole egg proteins starts coagulation at 68℃ while a
beaten eggwhite stars coagulation at 62℃. Accordind to McWilliams, “proteins in eggwhite
begin to coagulate at about 60℃ and lose ability to flow when temperature reaches
approximately 65℃ and that egg yolk proteins are somewhat more resistant to to the impact
of heat and begin coagulation at around 65℃ and cease to flow when temperature rises to
about 70℃” (2008, pg 279). Additionally, beaten whole egg coagulation temperature is
assumed to be the average the average of the coagulation temperstures of the eggwhite and
the egg yolk hence sapposed to be 62℃ by calculation. But according to Egg Safety Center
(2010), it is stated that the beaten whole egg starts coagulation at temperature around 68℃
which is ofcourse what was found as in the table. Heat is provides enerdy for the prtein
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molecules to unwind to secondary or primary structures and clump as they denature and
coagulate. Again, this coagulation is somewhat irrevesible.
Enzymatic browing of fruits and vegetables
Table 2.0. Enzymatic browning
Samples Treatment At room Bioled water Tap water vinegar
temperature
Irish potato cube Browning No browning Less No browning
browning
crashed More No browning Less No browning
browning browning
blanched Less to no No browning No browning No browning
browning
Apples Cut Broning No browning Less No browning
browning
As stated earlier in the introduction, enzymatic browning requires the prsence of the a
phenolic compound, an enzyme polyphenolase and presence of oxygen. So treating the fruits
with water and vinegar is inhibiting presence of oxygen and the enzyme respectvely. Because
vinegar is an acid, it therefore denatures proteins and because enzymes are protein in nature,
polyphenolase which is responsible for browning is also denatured. Hence in the table, there
is no browning under vinegar treatment. Tap water slightly contain some free oxygen which
will again be available for some browning to take place. This why most of the samples are
showing up less browning ecept for blanchen potato. Blanching potato denatures the
polyphenolase enzyme hence not available for any browning activity in tap water. Boiling
water reduces the amount of free oxygen in water and hence no browning is observable in all
the samples treated with bolied warter. Air has abundant free oxygen to accelerate the
enzymayic browning reaction. This is why all the samples shows browning under room
temperature. When fruits and vegetables are cut or bruised the phenols and the enzyme are
exposed to air and start reacting in the presence of oxygen to produce the brown coloured
products (Brown 2011).
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CONCLUSION
In conclustion, heat (temperature), beating (mechanical energy) and Ph (increasing acidity)
are the factors that cause protein to denature of which some denaturation are permanent or
reversible. For exmple beating an egg leads to a reversible denatuiration while heating and
adding acid brings about permanent denaturation. Denaturation and coagulation of proteins is
applicable in food processing and preparation as they work as thickenin agents in castards
and formation of cheese. Enzymatic browning occurs at different degrees in different
mediums. Fruits and vegetables readly go brown under exposure to air, show less to no
browning in boiled water and tap water and shows no browning in vinegar. Blanchin of
potato and some vegatables inhibits browning while crushing or cutting them into very small
pieces increases surface area for enzymatic browning. Henceforth, it could be recommended
to use vinegar, blanching and boiled water to contro enzymatic browning. But venegar may
still denature some impoortant proteins hence caution should be taken. Again, when
preparing fruits and vegetables avoid expexure to air and cut/chop them into pieces big
enough to reduce surface area for enzymatic browning.
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Reference
Brown A. (2011). Understanding Food; Principles and Preparation, (4th Ed). U.S.A;
Wadasworth Cengage Learning.
Debruyne L.K., Pinna K. & Whitney E. (2008). Nutrition and Diet Therapy, (8th Ed). U.S.A;
Brooks/Cole, Cengage Learning
Earl, B. & Wilford L.D.R. (2001). Chemistry, (2nd Ed). London; John Murray (Publishers)
Ltd.
Egg Safety Center,(2010). Coagulation of Eggs, Retrieved on 18/11/18, from
http://eggsafety.org/egg-safety/.
McWilliams, M, (2008).Foods; Experimental Perspectives, (6th Ed). New Jersey; Pearson
Education Inc.
