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Open Access | Page 10 |
Vol 3 | Issue 1 | Pages 10-18
Copyright: © 2022 Mae Chu M. This is an open-access arcle distributed under the terms of
the Creave Commons Aribuon License, which permits unrestricted use, distribuon, and
reproducon in any medium, provided the original author and source are credited.
Journal of Industrial Biotechnology
ISSN: 2578-6210
SCHOLARS.DIRECT
DOI: 10.36959/967/629
*Corresponding author: Dave Arthur R Robledo, Internaonal
Baccalaureate Diploma Program (IBDP), Saint Jude Catholic
School, Manila, Philippines
Accepted: April 01, 2022
Published online: April 04, 2022
Citaon: Mae Chu M, Robledo DAR (2022) An Invesgaon on
the Eects of Varying Temperatures on Gelan Denaturaon
in Response to Enzymac Reacons from Fruit Extracts. J Ind
Biotechnol 3(1):10-18
An Investigation on the Eects of Varying Temperatures
on Gelatin Denaturation in Response to Enzymatic
Reactions from Fruit Extracts
Megan Mae Chu1 and Dave Arthur R Robledo1*
1Internaonal Baccalaureate Diploma Program (IBDP), Saint Jude Catholic School, Manila, Philippines
Introduction
During holiday celebraons, my family would hold
gatherings in our home, and I would always help my mother
prepare some tradionally known desserts in the Filipino
culture, such as buko pandan (coconut salad with jelly) and
fruit salad. I was advised to use canned pineapples instead of
fresh ones because the laer contained more acve enzymes,
which “melted” the gelan in the salad. When we discussed
biomolecules in class, I became parcularly interested with
the topic, as it is relavely associated with biological processes
in the human body such as digeson and metabolism. Thus,
upon learning that gelan - a major component in fruit
salads - was made up of the biomolecule protein, I have
decided to invesgate the dierent factors that aected fruit
enzymes and how these catalysts subsequently aected the
breakdown of proteins. This would be demonstrated through
an experiment involving the use of materials that I was
already familiar with - fruits and gelan.
Research question
To what extent do varying temperatures (5°C, 55°C,
75°C, 100°C) aect the pH and the denaturaon of gelan in
response to exposure to fruit enzymes, such as bromelain,
papain, and amylases found in pineapple (Ananas comosus),
orange and green papayas (Carica papaya), and banana
(Musa acuminata), respecvely, by measuring the change in
pH (±0.01) and volume (±0.5mL)?
Hypotheses
For the change in volume
H1: Gelan denaturaon occurs the most in fruit samples
exposed to room temperature condion, as this temperature
provides an internal environment where enzymes funcon
most eecvely.
H0: There is no dierence observed in gelan denaturaon
when exposed to fruit samples from all temperature
condions.
For the change in pH
H2: The pH of all fruit samples decreases as the temperature
increases.
Research Article
Check for
updates
H0: Varying temperature condions do not have a
signicant eect on the pH of all fruit samples.
Background Information
Gelan is made up of an animal protein called collagen,
the most abundant protein in the human body found in
bones, muscles, and skin. It is a hard, insoluble, and brous
protein that are packed together to form long, thin brils,
which act as supporng structures to give the skin elascity
and strength [1].
With the presence of heat, the long chains of amino acids
coiled by the weak bonds in gelan begin to unwind. In this
process, the hydrogen bonds and non-polar hydrophobic
interacons are disrupted by heat due to kinec energy.
With an increase in heat, the molecules move rapidly and
violently, disrupng the bonds in the process. The process of
denaturaon in secondary and terary proteins is illustrated
in Figure 1, depicng how the structures are destroyed into
random coils under heated condions [2].
Aer some me, the gelan cools down and solidies;
in this process, the collagen reforms, and the inial protein
structure is no longer achieved due to the disrupon of the
normal alpha-helix and beta sheets from denaturaon [2].
Ergo, the protein now reforms in a random, tangled structure.
Water is trapped in the middle of the long chain proteins in
the process, turning the liquid state of protein into a semi-
solid mass. The protein structure of a semi-solid mass remains
the same even under room temperature.
Electronic copy available at: https://ssrn.com/abstract=4085991
Citaon: Mae Chu M, Robledo DAR (2022) An Invesgaon on the Eects of Varying Temperatures on Gelan Denaturaon in Response to
Enzymac Reacons from Fruit Extracts. J Ind Biotechnol 3(1):10-18
Mae Chu M. J Ind Biotechnol 2022, 3(1):10-18 Open Access | Page 11 |
as it breaks protein down into smaller fragments of amino
acids and pepdes [8]. Based on the research of Babu (2019)
[9], papain is opmally acve between a pH of 5 to 7.5 at
70 - 90°C.The proteolyc enzymes bromelain and papain
were compared side-by-side based on their use as a meat
tenderizer for squid (Loligo vulgaris) in Gokoglu, et al. (2017)
[10], and results showed that beer results were obtained
from papain compared to bromelain.
Lastly, another fruit that contains an abundant number of
enzymes is banana (Musa acuminata). The fruit is composed
of amylases, which aid in the breakdown of complex
carbohydrates into easier to absorb simple sugars in the body
[11]. The maximum acvity of amylase was found to be within
the pH range of 6 - 7, and it is most acve at 62°C according to
Kinsella & Mao (2006) [12].
Factors Aecting Enzyme Activity
While it is true that enzymes work best in an internal
environment that mimics the human body temperature
(37°C), there sll exist factors that inuence enzyme acvity.
Enzymes have an opmum temperature at which they can
work best. It is observed in the illustraon in Figure 2 that
as the temperature gradually increases, the rate of enzyme
acvity also increases. This is true, but only unl the peak of
opmum temperature is reached. At opmum temperature,
the enzyme works at its fullest potenal; however, as
presented in the curvilinearity of the graph, the rate of enzyme
acvity connuously declines as the temperature increases
Enzymes in Fruits
Catalyc enzymes are known to accelerate the breakdown
of pepde bonds found in the link of long chain proteins by
lowering the acvaon energy. With this, the links of long
chain proteins break loose into smaller proteins. The trapped
water from the bonds is then released, and the soluon
returns into liquid. This explained why the gelan in the
experiment became liqueed as it came in contact with the
fruit samples.
Fruits are an abundant source of enzymes that aid in the
digeson of ingested protein and carbohydrates. According
to Kaur, et al. (2014) [3], bromelain is a sulydryl enzyme
found in the fruit and stem of pineapple, renowned for its
role in the promoon of remedying digesve disorders and
as a natural meat tenderizer. It is an essenal component in
the proteolysis and digeson of protein in the human body.
The proteolyc enzyme funcons by aacking the internal
pepde bonds of the protein chain [4]. To eecvely funcon,
the opmum pH of 7.1 must be reached. It is most stable
from the pH ranges of 3.9 - 4.2, and its opmum temperature
is 55°C [5].
Aside from the diversity of funcons bromelain oers,
its proteolyc acvity is known to be 10 mes higher than
that of papain [6]. Papain is another proteolyc enzyme
obtained from the latex of raw papaya fruit (Carica papaya)
[7]. Similar to the funcon of bromelain as a meat tenderizer
and a digesve agent, papain also boasts the same abilies,
Figure 1: The process of protein denaturaon.
Electronic copy available at: https://ssrn.com/abstract=4085991
Citaon: Mae Chu M, Robledo DAR (2022) An Invesgaon on the Eects of Varying Temperatures on Gelan Denaturaon in Response to
Enzymac Reacons from Fruit Extracts. J Ind Biotechnol 3(1):10-18
Mae Chu M. J Ind Biotechnol 2022, 3(1):10-18 Open Access | Page 12 |
and exceeds the opmum point. At this part, the inial shape
of the acve site of the enzyme becomes disrupted when
exposed to extreme temperatures, disabling the aachment
of a substrate to the enzyme’s acve site.
Aside from temperature, enzymes also respond to
changes in pH. An enzyme can undergo conformaonal
changes when a change in pH is induced in its environment.
This is explained through the charge in amino acids. Within
the enzyme are posively- and negavely- charged amino
acids that are aracted to one another. The shape of the
enzyme is aributed to the aracve forces between the
amino acids. When the pH is changed, the charges in the
amino acids are aected; thus, the amino acids are no longer
aracted to one another. As a result, the enzyme undergoes
a conformaonal change - most oen permanently - which
produces a denatured enzyme [13]. Similar to temperature,
the eect of pH depicts the same trends in the rate of
enzyme acvity. However, there is also an opmum pH in the
graph at which the enzyme is likewise able to funcon most
eecvely, yet this is only true to a certain extent because the
rate of enzyme acvity encounters a decline once the enzyme
exceeds its opmum pH measurement.
Methodology
Variables
Independent variable: Varying temperatures (5°C, 55°C,
75°C, 100°C)
Dependent variables: Change in volume (gelan
denaturaon) (mL) (±0.5mL), change in pH (±0.01)
Controlled variables
1. The volume of the gelan (mL) was evenly distributed
to ve empty beakers with a measurement of 60 mL in each.
2. The mass of the fruits (g) was measured on a digital
weighing scale to be exactly 40g. This was done to prevent
unequal measurements in gelan denaturaon. It was more
reasonable to measure the fruits in mass (g) than in volume
(mL) since the fruits had dierent consistencies aer being
blended - some were in chunks and some were more liqueed.
3. The room temperature condion was the control
group in this experiment. It did not undergo any extreme
temperature condions and remained at room temperature
at all mes. It was placed far away from an external heat
source. Hence, no changes were observed in the dependent
variables under this condion.
4. To prevent unequal measurements in gelan
denaturaon, the duraon of fruit exposure in gelan (hrs)
was controlled. Once the fruit sample was poured into the
gelan, a mer was set at 1.5 hours.
5. The me of temperature exposure of fruit samples
(mins) remained consistent throughout the experiment; all
mers lasted for twenty (20) minutes.
6. To ensure that the gelan’s response to fruit enzymes
was not varied in all trials, the gelan mix used in all samples
- Knox unavored gelan - was standardized.
7. The fruit samples for all trials under each condion
were prepared using the same fruit.
Materials
• 4 fruits (pineapple, orange papaya, green papaya, banana)
• 6 boxes of Knox unavored gelan
• 5 pieces of 100 mL beakers (±0.5mL)
• Oven mis
• Mixing spatula
• Chopping board
• Kitchen knife
• Mobile phone as a mer (±1s)
• 4 in 1 pH meter (±0.01)
*This brand was chosen due to its automac calibraon
feature.
• Tanita digital weighing scale (±0.1g)
*This brand was chosen due to its availability in the
kitchen.
Figure 2: The eects of temperature and pH on the rate of enzyme acvity.
Electronic copy available at: https://ssrn.com/abstract=4085991
Citaon: Mae Chu M, Robledo DAR (2022) An Invesgaon on the Eects of Varying Temperatures on Gelan Denaturaon in Response to
Enzymac Reacons from Fruit Extracts. J Ind Biotechnol 3(1):10-18
Mae Chu M. J Ind Biotechnol 2022, 3(1):10-18 Open Access | Page 13 |
• Philips blender
• Oven
• Refrigerator
Preliminary Set-up
1. The inial mass of the beakers containing gelan was
pre-weighed to measure the mass of the fruit juice when it
was later placed.
2. The gelan mix was prepared separately in a large bowl,
then poured into ve (5) empty beakers at 60 mL. These were
placed inside the refrigerator to solidify for three (3) hours.
3. While waing for the gelan to solidify, the fruit was
sliced into smaller pieces then placed in a blender unl it
reached a liquid consistency. Following this procedure, the
inial pH of the fruit juice was measured.
Procedure
1. Once taken out of the refrigerator, the gelan samples
were placed on the table at room temperature to cool down
for thirty (30) minutes. This was to ensure that the cold
temperature, which the samples were previously exposed to,
did not become a confounding variable in directly aecng
the gelan denaturaon.
2. While waing for the gelan to cool, the selected
fruit was exposed to its respecve condion for twenty (20)
minutes. In the 5°C condion, the sample was placed inside
the freezer; for the following condions, the samples were
placed inside the pre-heated oven.
3. The fruit was le to cool down for thirty (30) minutes
to ensure that its temperature did not have a direct eect on
the gelan denaturaon. The pH of the fruit sample was then
measured.
*The purpose of exposing the fruit to varying temperatures
was solely for fruit enzyme denaturaon; thus, there was no need
to keep the temperatures constant throughout the experiment.
4. Next, the fruit juice was poured into the surface of the
gelan in ve (5) beakers. Each beaker was representave of
the ve (5) trials under the temperature condion the fruit
was exposed to.
5. A mobile phone was used to set a mer for 1.5 hours.
6. Aer the alloed me, the change in volume (mL) was
measured.
7. Aer compleng the experiment for the rst ve (5)
trials, the same procedure was repeated for the following
trials. Each fruit consisted of twenty-ve (25) trials and there
were ve (5) condions in total.
Figure 3: The set-up of gelan with banana under 75°C
Figure 4: The set-up of gelan with banana under 75°C
aer 1.5 hours
Risk assessment
Safety Concerns: Since most of the samples needed to
be exposed to high temperatures, oven mis were worn
when the oven was accessed to prevent geng burns. The
assistance of a family member at home was needed while
using the oven and using the knife to cut the fruits to ensure
that all safety precauons were followed.
Environmental concerns: The samples were disposed of
in the trash bin to lessen the chances of food contaminaon.
Ethical concerns: There were none surrounding this
experiment.
Presentation of Raw Data
Table 1: Change in pH in all fruit samples under varying
temperature condions aer 1.5 hours
Table 2: Change in gelan volume (mL) in all fruit samples
under varying temperature condions aer 1.5 hours
Observations
From temperatures 5°C to 75°C in all fruit samples, it was
seen that the fruits penetrated through the gelan surface
and turned the aected part into a liquid-like consistency aer
the exposure. Figure 5 illustrated the complete disintegraon
of gelan in response to the eect of pineapple, as seen in the
color transion of gelan from a translucent appearance to
an opaque, bright yellow. In Figure 6, there was a producon
of eervescence in green papaya aer a prolonged me,
showing an indicaon of the release of oxygen from water
due to the breaking of the hydrogen bonds in gelan caused
by the catalyc enzyme bromelain.
Processed data
To solve for the average changes in volumes and pH, this
equaon was used:
sin
(5) (5) (4) (7) (10)
55
5 6.20
sum of all the change the trials
xtotal number of trials
banana C condition
banana C condition
=
+++ +
=
=
Table 3: Average change in gelan volume (mL) in all fruit
samples under varying temperature condions aer 1.5 hours.
Figure 3: The set-up of gelan with banana under 75°C.
Figure 4: The set-up of gelan with banana under 75°C aer 1.5
hours.
Electronic copy available at: https://ssrn.com/abstract=4085991
Citaon: Mae Chu M, Robledo DAR (2022) An Invesgaon on the Eects of Varying Temperatures on Gelan Denaturaon in Response to
Enzymac Reacons from Fruit Extracts. J Ind Biotechnol 3(1):10-18
Mae Chu M. J Ind Biotechnol 2022, 3(1):10-18 Open Access | Page 14 |
Table 1. Change in pH in all fruit samples under varying temperature condions aer 1.5 hours
Table 2. Change in gelan volume (mL) in all fruit samples under varying temperature condions aer 1.5 hours
Table 3. Average change in gelan volume (mL) in all fruit samples under varying temperature condions aer 1.5 hours
Table 4. Average change in pH inall fruit samples under varying temperature condions aer 1.5 hours
Electronic copy available at: https://ssrn.com/abstract=4085991
Citaon: Mae Chu M, Robledo DAR (2022) An Invesgaon on the Eects of Varying Temperatures on Gelan Denaturaon in Response to
Enzymac Reacons from Fruit Extracts. J Ind Biotechnol 3(1):10-18
Mae Chu M. J Ind Biotechnol 2022, 3(1):10-18 Open Access | Page 15 |
Figure 5: Set-up of gelan with pineapple under 55°C aer 1.5
hours.
Figure 6: Set-up of gelan with green papaya under 55°C aer
12 hours.
0
0.1
0.2
0.3
0.4
0.5
0.6
5℃ 55℃ 75℃ 100℃
Change in pH (±0.1)
condi�ons
Banana Pineapple Orange Papaya Green Papaya
Figure 7: Change in gelan volume (mL) in all fruit samples under
varying temperature condions with standard error.
0
5
10
15
20
25
30
5℃ room
temp.
55℃ 75℃ 100℃
change in volume (±0.5mL)
condi�ons
Banana Pineapple Orange Papaya Green Papaya
Figure 8: Change in pH in all fruit samples under varying
temperature condions with standard error.
Table 4: Average change in pH inall fruit samples under
varying temperature condions aer 1.5 hours.
In Figure 7, it was demonstrated that at higher
temperatures, enzymac acvity became weaker in
denaturing gelan because the rate of enzymac acvity
declined aer the opmum temperature, for each
temperature has been surpassed. Based on the results of
the experiment, the condion with the most acve enzymes
was at room temperature, and the lowest was at 5°C. Among
all the fruits, pineapple had the most noceable eects on
gelan denaturaon; on the contrary, the fruit with the lowest
enzymac acvity was in banana. In Figure 8, the trends
seemed inconsistent in all fruit samples, as some increased
then decreased and vice versa, whereas one connuously
decreased across the increasing temperatures. This clearly
illustrated that pH did not decrease as temperature increased.
Opmally acve at 55°C, papain from papaya at 70°C to
90°C, and amylases from banana at 62°C, all the fruit samples
worked most eecvely at room temperature. Bromelain
was said to be most stable between 3.9 to 4.1 pH, and this
was proven true in the study, as the pH of pineapple in all
condions approximately fell within the range. Between
orange and green papayas, the laer resulted in a more
denatured gelan than orange papaya. Papain was expected
to be opmally acve between 5 to 7.5 pH, and this was
conrmed in the study, as the pH measurements of the
samples in all condions t within the supposed range. In
banana, its pH ranged from 4.5 to 4.9, which did not coincide
with the literature values wherein amylases were said to be
opmally acve between 6 to 7 pH.
Statistical Test
The error bars in Figures 7 and Figure 8 suggested that
some data were more spread around the mean than others.
Hence, a Shapiro-Wilk test was used to examine whether
the samples (including outliers) were normally distributed
to determine the appropriate stascal test to be used later.
The tests on the average change in volume and the average
change in pH were ran on Microso Excel.
Table 5 showed that all samples were normally distributed,
where the values of W were found to be closer to 1, a predictor
of the data being normally distributed. Aside from this, it was
also observed that the p-value of all condions was greater
than the chosen σ level 0.05, hence the data being normally
distributed. The same results were observed in Table 6 where
the data was found to be normally distributed in all groups:
Electronic copy available at: https://ssrn.com/abstract=4085991
Citaon: Mae Chu M, Robledo DAR (2022) An Invesgaon on the Eects of Varying Temperatures on Gelan Denaturaon in Response to
Enzymac Reacons from Fruit Extracts. J Ind Biotechnol 3(1):10-18
Mae Chu M. J Ind Biotechnol 2022, 3(1):10-18 Open Access | Page 16 |
Due to the data being normally distributed, a one-way ANOVA
test was conducted using StatPlus to determine its signicance.
This stascal test was chosen because there were numerous
samples present, and the study only had one independent
variable. Tables 7 and Table 8 below showed the results from
the test on the change in volume (mL) and the change in pH.
Due to the equal number of samples present in each group, the
homogeneity assumpon for the variances was ignored.
With an obtained p-value of 0.00338, which was less than
the signicance level of 0.05, Table 7 demonstrated that the
data was signicant. Based from Table 7, the 100°C condion
had the lowest standard deviaon of 1.95 among the ve
groups, indicang that the dierences in the condion were
closer to one another. The room temperature condion
resulted in a standard deviaon of 6.80, which meant that
the dierences were larger in this condion. The same was
observed in the mean - the 100°C condion had the lowest
mean (3.50) and the room temperature condion had the
highest mean (18.4), indicang that the enzymes were able
to work best under room temperature. With the data being
signicant at p < 0.05, and pineapple having the highest mean
in the change in volume, the rst alternate hypothesis for the
change in volume was accepted. This result supported the
noon that enzymes work best in an internal environment
because it shares a close resemblance to the human
body temperature (37°C), wherein they play a vital role in
maintaining biological processes. It was therefore understood
that pineapple, at room temperature, resulted to the most
prominent disrupon of protein chains in gelan, which led to
releasing trapped water from the bonds. This explained why
its outcome produced the most liquid-like consistency as it
came in contact with the fruit (See Figure 5).
5℃ (±0.5mL) room temperature 55℃
(±0.5mL)
75℃
(±0.5mL)
100℃
(±0.5mL)
W 0.945775 0.980980 0.964103 0.921791 0.944664
p-value 0.307459 0.946117 0.628637 0.107275 0.293246
Table 5. Shapiro-Wilk Test on the average change in volume in gelan (mL) in all fruit samples under varying temperature condions aer
1.5 hours
5℃ (±0.1) room temperature 55℃ (±0.1) 75℃ (±0.1) 100℃ (±0.1)
W 0.944206 NA 0.782638 0.904323 0.875665
p-value 0.902342 NA 0.123842 0.626649 0.440688
Table 6. Shapiro-Wilk Test on the average change in pHinall fruit samples under varying temperature condions aer 1.5 hours
Table 7. ANOVA test onthe change in volume (mL) in all fruit samples under varyingtemperature condions
Table 8. ANOVA test onthe change in pH in all fruit samples under varyingtemperature condions
Electronic copy available at: https://ssrn.com/abstract=4085991
Citaon: Mae Chu M, Robledo DAR (2022) An Invesgaon on the Eects of Varying Temperatures on Gelan Denaturaon in Response to
Enzymac Reacons from Fruit Extracts. J Ind Biotechnol 3(1):10-18
Mae Chu M. J Ind Biotechnol 2022, 3(1):10-18 Open Access | Page 17 |
The obtained p-value of 0.42462 provided in Table 8
proved that the data was insignicant, as it was greater than
the signicance level of 0.05. Among the ve condions, the
75°C condion had the lowest standard deviaon of 0.0624,
indicang that the dierences in the condion were closer
to one another. The 55°C condion had the highest standard
deviaon of 0.196, which meant that the dierences in the
condion were larger compared to the former. The same was
observed in the mean - the 75°C condion had the lowest
mean (0.113) and the 55°C condion had the highest mean
(0.16), which meant that the largest decrease in pH values was
observed in the laer. There were no observed changes in pH
at room temperature condion because it was not exposed
to any extreme temperature condions. For the reason that
the data was not signicant at p < 0.05, the null hypothesis for
the change in pH failed to be rejected. Thus, it was inferred
that varying temperature condions did not have a signicant
eect on the pH of all fruit samples.
Following this, the change in pH across the temperature
condions was graphed to beer understand the trend
present in the variables:
An inverse relaonship between pH and temperature
with a moderate correlaon of determinaon (R2 = 0.5156) is
presented in Figure 9. From 5°C to room temperature, there
was an increase in pH; as the temperature reached 55°C,
the change in pH gradually decreased, and fairly increased
towards 75°C. At 100°C, there was a drasc decline as the
pH connued to decrease to -1.1275. Due to this, it was
deduced that temperature did not have a signicant eect on
pH, but they shared an inverse relaonship and a moderate
correlaon.
Evaluation
Conclusion
Based on the results of this study, the enzymes in all
the fruit samples worked most eecvely under room
temperature, as this mimicked the normal body temperature
at which enzymes become opmally acve. Among all the
fruits involved, pineapple demonstrated the most acvity
in denaturing gelan, and banana with the least acvity.
A reason for the occurrence that bromelain resulted to
a higher denaturing capability than amylases was that
bromelain was a proteolyc enzyme known to digest
proteins, whereas amylases were instead involved in the
breakdown of carbohydrates into simple sugars. Moreover,
at room temperature, it was implied that the enzymes
remained unaected as they did not denature or undergo
any conformaonal changes at this condion due to the
absence of extreme temperatures, hence the gelan became
more resistant to the acon of enzymes. Due to this and with
the data being signicant, the rst alternave hypothesis for
the change in volume was conrmed by the data, insinuang
that gelan denaturaon occurred the most in fruit samples
exposed to room temperature condion, as this temperature
provided an internal environment where enzymes funcon
most eecvely.
In measuring the change in pH, temperature did not
have a signicant eect on pH, so a strong relaonship
between the variables could not be established. Among
the temperature condions, the highest change in pH was
found to be at 55°C and the lowest at 75°C. Among all the
fruit samples, the highest change in pH occurred in orange
papaya; the lowest change, in banana. Hence, the second null
hypothesis for the change in pH was refuted by the data. This
suggested that varying temperature condions did not have
a signicant eect on the pH of all fruit samples. However,
it was sll worth nong that temperature and pH shared a
moderate inverse relaonship (R2 = 0.5156), as demonstrated
in Figure 9. To explain this, as the temperature increased, the
subsequent molecular vibraons caused ionizaon to occur,
hence more hydrogen ions were formed. As a result, the pH
dropped (“How Does Temperature Aect pH?”), leading to
an inverse relaonship. Ulmately, the results showed that
temperature sll played a part in the measurement of pH.
Strengths
The experiment certainly had strengths due to it having
several controlled variables in the procedure. There were
5 trials used in 4 fruit samples under each temperature
condion, which ensured a more consistent representaon
of results. Although the measuring devices used in the
experiment could not guarantee a high level of accuracy
due to the presence of measurement errors, the uncertainty
measurements for all apparatuses used in the experiment
and the standard error in Figures 7 and Figures 8 sll helped
improve accuracy. Another strength would be the use of
standardized samples from Knox unavored gelan, which
stayed consistent throughout all the trials. This ensured that
the gelan in all samples reacted the same way to the fruit
enzymes.
Limitations
A limitaon to the procedure included the lack of
appropriate measuring apparatuses due to it being a home-
based experiment. A kitchen scale was used to weigh the
fruit samples instead of a laboratory weighing scale, which
could only measure the mass of the samples unl the second
decimal place. Using the laer would have lessened random
and measurement errors due to its precision in detecng
the mass of the sample unl the smallest decimal. Also,
it was worth nong that although the fruit samples came
from the same batch, their ripeness was not assessed or
-1.5
-1
-0.5
0
5℃
room temp.
55℃
75℃
100℃
change in pH (±0.1)
condi�ons
y = - 0.2103x + 0.3243
R² = 0.5156
Figure 9: Relaonship between pH and temperature in all fruit
samples under varying temperature condions.
Electronic copy available at: https://ssrn.com/abstract=4085991
Citaon: Mae Chu M, Robledo DAR (2022) An Invesgaon on the Eects of Varying Temperatures on Gelan Denaturaon in Response to
Enzymac Reacons from Fruit Extracts. J Ind Biotechnol 3(1):10-18
Mae Chu M. J Ind Biotechnol 2022, 3(1):10-18 Open Access | Page 18 |
measured before the experiment. Ethylene producon in
fruits causes ripening, and in this process, new enzymes are
produced [13]. This poses a hindrance in acquiring more
accurate data because the fruits may have been at dierent
levels of ripeness, aecng the proteolyc acvity of the
enzymes in protein denaturaon in the gelan. This may have
consequently resulted to the presence of outliers in the data,
ergo aecng its signicance.
Extensions
For future studies, experimenng in a laboratory seng
is recommended due to access to appropriate apparatus.
This would allow a more controlled environment for the
variables, and using measurement tools with smaller
uncertaines would lessen the percentage error in data.
Because the temperature condions used in the experiment
were pre-set temperatures in the oven, the enzymac
acvity under condions closer to body temperatures (for
example, 35°C to 40°C) was not tested. This would have been
an interesng take on understanding beer the chemical
processes occurring inside our bodies. Addionally, the
gelan denaturaon should occurat constant temperature,
unlike in this study, where only a 20-minute exposure was
performed in denaturing fruit enzymes. Addionally, the
selecon of fruit samples may be expanded to a wider variety
of fruits whose enzymes may also aid in digeson. Beer yet,
it would be a worthwhile opportunity to test the proteolyc
properes that uncommon fruits may possibly contain. Most
importantly, it would be advisable to quantavely measure
the ripeness level of the fruit samples beforehand to reduce
possible confounding variables in the experiment. Perhaps
this could be done by using a spectrophotometer to inspect
the colored pigment molecules or light absorpon intensity
of the fruit’s skin as a means of determining its age.
References
1. McIntosh J (2017) What is collagen, and why do people use it?
Medical News Today.
2. Denaturaon of proteins.
3. Kaur T, Kaur A, Grewal R (2015) Kinecs studies with fruit
bromelain (Ananas comosus) in the presence of cysteine and
divalent ions. J Food Sci Technol 52: 5954-5960.
4. Wijeratnam S (2016) Pineapple. Encyclopedia of Food and
Health.
5. Bromelain. Creave Enzymes.
6. Bromelain. Naonal Center for Complementary and Integrave
Health.
7. Arvanitoyannis I, Varzakas T (2008) Fruit/Fruit juice waste
management: Treatment methods and potenal uses of treated
waste. Waste management for the food industries.
8. McDermo A (2018) 6 ways to use papain. Health line.
9. Babu K (2019) The dyeing of silk. In. Papain. (2nd Edn). Science
Direct.
10. Gokoglu N, Ucak I, Yatmaz H, et al. (2017) Eect of bromelain
and papain enzymes addion on physicochemical and
textural properes of squid (Loligo vulgaris). Journal of food
measurement and characterizaon 11: 347-353.
11. Oswald C (2018) Digesve Enzymes: Amylase, protease, and
lipase. Integrave Therapeucs.
12. Kinsella JE, Mao WW (1981) Amylase acvity in banana fruit:
Properes and changes in acvity with ripening. Journal of Food
Science 46: 1400-1403.
13. What happens in cells and what do cells need? Factors aecng
enzyme denaturaon.
14. Koning R (1994) Fruit growth and ripening.
Copyright: © 2022 Mae Chu M. This is an open-access arcle distributed under the terms of
the Creave Commons Aribuon License, which permits unrestricted use, distribuon, and
reproducon in any medium, provided the original author and source are credited.
SCHOLARS.DIRECT
DOI: 10.36959/967/629
Electronic copy available at: https://ssrn.com/abstract=4085991
