The relationship of catalase and temperature; Effect

of salinity in the germination of mung beans

Dy, Chloe1; Garcia, Ana Marie Leoni1; Piaduche, Andrey Marie1; Rosales,
Mary Chris1

[1] Science, Technology, Engineering, and Mathematics Student, Senior High School Department,
Polytechnic University of the Philippines

Abstract

Nearly all living things possess catalyst or the substances within them that speed up chemical reactions
and processes. Enzymes are macromolecular biological catalysts that accelerate chemical reactions that
take place within cells. This experiment utilizes catalase as the enzyme and uses hydrogen peroxide as the
substrate that it breaks down. Also, the action of protein was tested on sliced potato, carrots and liver in
different test tubes that was placed in cold and hot water. In addition, enzyme should be able to function
in the conditions that surround it in the tissue it's supposed to function in, and some of it can be damaged
under certain conditions, such as heat and a damaged enzyme will no longer work to catalyze a chemical
reaction. The outcome of the experiment shows that each kind have different rate of chemical reactions
which results in the products of oxygen gas (O2) and liquid water (H2O), and the action of protein in
breaking down hydrogen peroxide was faster in liver.

Water is one of the essential components that are needed for plants to live and grow, but does the
composition of the water make any difference and if so, what is the ideal? The answers to these questions
are undeniably a big step in the agricultural industry of the country, allowing it to grow more and better
crops. By this experiment, the researches wanted to know which concentration of salt would be
exemplary for the germination of the mung bean. The researchers hypothesized that the composition or
salinity of water would affect (positive or negative) the growth of the Mung beans, and one of those
formulas (particularly NaCl solution or regular water) would be ideal for the growth of the plants.
Previous researchers hypothesized that Germination decreases along with increasing NaCl concentrations
(Dept. of Botany, Scottish Church College, India, 2014).

Keywords: Enzyme, Catalase, Salinity

Introduction

Enzymes are protein catalysts, and a catalyst is a temperature, and then they will begin to
substance that speeds up the rate of a chemical denature and become inactive (Aronson, Fraser,
reaction without being used up. Enzymes Smith, 1956)
catalyze most if not all chemical reactions that
There are three essential things that plant need in
take place within living cells (Morris, 2011).
order to grow. These are sunlight, water and
This experiment utilizes catalase as the enzyme
nutrients, if you were to alter any of these
and uses hydrogen peroxide as the substrate that
necessities the plant will be the one who is most
it breaks down. The experiment had two parts to
likely to suffer. The uptake of water from the
it. In the first part, the hypothesis was that as
roots, through the xylem and into the rest of the
substrate concentration increased, so would the
plant is an essential part of a plants ability to
reaction rate until all active sites are bound. In
photosynthesize. Also, water is necessary to
the second part, it was hypothesized that as
create cell tension, which gives plants their
temperature increased, so would the reaction rate
form. Lastly, some minerals, such as calcium,
until the enzyme is denatured. The purpose of
are stored in water and the plant can only access
this experiment was to observe how varying the
these minerals by up taking water (Graham
temperature of the catalase and varying the
2006). Due to the extreme importance of water
substrate concentration affected the reaction rate
in the life of a plant, we decided to investigate
of O2. The null hypotheses were that as the
the effect water quality has on plant growth.
substrate concentration increased, so would the
Salinity refers to the salt content of any given
reaction rate until all the active sites were bound,
system. According to Pichu Rengasamy,
and that as the temperature of the catalase
“Salinization is one of the most devastating
increased, so would the reaction rate until the
forms of land degradation threatening food
enzyme was denatured. The alternative
production worldwide” (March,2008). It has
hypotheses were that the varied substrate
always been evident that salinity has its negative
concentrations and varied temperature would
effect on the growth of many plats and that fact
have no effect on reaction rates. The catalase
is rooted at the bases of the researchers’
speeds up the reaction proportional to the
experiment. The Independent variable in this
amount of substrate added. Once the active sites
experiment is the percentage of salinity in the
of the catalase are filled, it will no longer
salt solution. The dependent variable is the
function to catalyze the reaction. Likewise,
growth of the mung beans.
enzymes work at very specific temperatures, so
they will catalyze a reaction up to a certain
Methodology
1.0 Catalase Experiment
For each test that was conducted, the researchers
used 1cc syringe, test tube, cutter, circular
plasticware, hot and cold water, and especially,
the carrots, potato, liver, and hydrogen peroxide
as some of the independent variables. The
researchers divided the carrots, potato, and liver
into same sizes and place it, individually, at the
bottom of the of the test tube. While the test tube
was dipped in hot and cold water, respectively,
hydrogen peroxide was added within. The
reaction produced a bubbling effect and the
height of the bubbles were measured.
Researchers observed the factors that affect the
enzyme.
2.0 Salinity Experiment
To test the hypothesis, the researchers have 4
sets of mung beans with different salinity of
water. This experiment used circular
plasticware, tissue paper, water, and salt. The
salt was used by the researchers to produce
0.6%, 0.9%, and 1.2% salinity of water. First,
the researchers placed the tissue inside of the
circular plasticware and cut holes on the lid,
afterwards, the mung beans were set inside the
plasticware. When everything was settled, the
researchers watered each of the plasticware with
0.6% salinity, 0.9% salinity, 1.2% salinity, and
with regular water.
For seven (7) consecutive days, the researchers
watered it each day with its own water
composition and monitor its growth. The
researchers have monitored the mung beans'
characteristics and factors that may affect its
growth.
Results and Discussion
1.0 Catalase Experiment
Table 1: The reaction’s height of the three
variables, submerged in a cold water.
Test Tube
1 2 3
Carrot 0.5 0.3 0
Potato 1 0.5 1
Liver 2.5 4.4 4
The table number 1 interpret the height of the
reaction of the three variables, which is the
carrot, potato and liver that was submerged with
hydrogen peroxide in a test tube, while the test
tube was also submerged in a cold water. As for
the first variable; carrot, in its first test the
reaction is quiet low and the catalase is slower to
react that measures 0.5cm only, and the foam
became only visible after a couple minutes, there
was only a slight different in the result as we
conduct the second test that measures 0.3cm
and the third test with no foam or reaction to
measure. The second experiment of measuring
the reaction was conduct with the use of potato
as the variable, as the test goes by the
measurement of potato’s catalase reaction was
maintained from the first test to third test that
measures 1cm, it shows that there’s only a slight
difference in the second test where the height of
the reaction measures 0.5cm, lastly is the
experiment using the liver which managed to conducted experiment the liver made the most
show the highest measured reaction and quickest quickest reaction when it is submerged with
variable to react in the hydrogen peroxide while hydrogen peroxide inside the test tube, it
its test tube was being placed in a cold water, in managed to have a height reaction of 6cm in the
the first test its foam measures 2.5cm that first and second test, and it increases more in the
increases in the second test that measure 4.4cm third test where it measures 7.1cm.
and decrease a little in the third test having 4cm.
Three kind of protein was used to measure it's
Table 2: The reaction’s height of the three catalase reaction submerged with hydrogen
variables, submerged in a hot water. peroxide, the experiment was conducted three
times to see if any difference will be shown in
Test Tube the result, first experiment conducted is the
1 2 3 reaction of liver's catalase submerged with
Carrot 0 0.5 0.2
hydrogen peroxide, the observation is that when
Potato 0.5 1.5 0.5
the bigger the surface area the quicker the
Liver 6 6 7.1
reaction will be.

The table number 2 shows the height of the First the liver, its quick reaction is because of its

reaction of the three variables, which is the more particles that was exposed to the hydrogen

carrot, potato and liver that was submerged with peroxide therefore the liver that is ground up

hydrogen peroxide in a test tube, while the test will be the quickest reaction. In the conducted

tube was also submerged in a hot water. As for experiment only a small part of the liver was

the first variable; carrot, at first test it shows no exposed hence the test tube was too small for the

reaction at all, in the second test it made a slight liver that became the cause why there's only a

reaction where the reaction height measures 0.5 little amount of foam formed; in regards to the

cm and decreases again in the third test where reaction of liver, but among the three protein,

the height reaction only measures 0.2cm, the the liver has the highest foam and reaction and

second variable used in the experiment is the quickest to react in the hydrogen peroxide, in

potato, it also made the slightest difference because the liver has more catalase than the two,

between the three test while measuring its The variables that affect the decomposition of

reaction height, where the first and third test hydrogen peroxide are, The temperature of the

measures 0.5cm while the second test measures liver, The surface area of the liver, The pH of

1.5cm that made the most visible reaction the hydrogen peroxide and The concentration of

among the three test, the third and last variable the enzymes, where in the experiment the liver

to be observed is the liver, among all the

has a quiet more foam reaction in hot water than enzymes are found in living cells and are made
in the cold water, of proteins. As enzymes are made of protein if
the temperature is too high it becomes damaged.
While in potato, when the sliced potato is placed
The collision theory states that particles need
in a test tube and submerged with a hydrogen
enough kinetic energy for a reaction to happen.
peroxide solution, bubbles also suddenly
appeared which only shows that oxygen is being To say the least almost similar variables among
evolved, but it's the slowest to react among the the three trigger the reaction to happen. There is
three protein, it took a couple of minutes before a slowing or stopping down of the catalase
to actually see the bubbles, that also the mostly because of the drastic change in the
temperature plays a big part in the experiment enzymes environment which cause by the
where in the potato has much more foam enzyme and substrate to bind or to not bind
reaction in cold water, which is the total of correctly.
opposite in the liver experiment,
2.0 Salinity Experiment
In the experiment the key factors that possibly
affect the result of the potato submerged with
hydrogen peroxide are the temperature of the
surroundings and test tubes, The pH value of its
surroundings, The substrate volume,
Concentration of the H202, (Substrate mass),
Surface area of the potato, (surface area of the
Figure 1: The effect to the growth of mung bean
active site) and the time, lastly is the carrot's watered by a regular water; Day 1.
catalase reaction in hydrogen peroxide,
Figure 1 show no progress to the growth of the
In the experiment the carrot has the slowest
mung bean watered by regular water on it first
reaction among the three, and the observation is
day.
that when the temperature increased more foam
was produced, the carrot reaction was more
visible in the hot water, The temperature did
affect the catalase in the carrot cells when the
hydrogen peroxide (H2O2) was added. The
enzyme breaks down the H2O2 to produce water
and oxygen which made the foam. Most of the
foam was produced in the range of 0-.5 cm, Figure 1.1: The effect to the growth of mung bean
watered by a 0.6% salinity of water; Day 1.
Figure 1.1 show a little progress to the growth of
the mung bean watered by a 0.6% salinity of
water on its first day.

Figure 2: The effect to the growth of mung bean

watered by a regular water; Day 2.

Figure 1 still show no progress to the growth of

the mung bean watered by regular water on its
second day.
Figure 1.2: The effect to the growth of mung bean
watered by a 0.9% salinity of water; Day 1.

Figure 1.2 show no progress to the growth of the

mung bean watered by a 0.9% salinity of water
on its first day.

Figure 2.1: The effect to the growth of mung bean

watered by a 0.6% salinity of water; Day 2.

Figure 2.1 shows a little progress to the growth

of the mung bean watered by a 0.6% salinity of
water on its first day where a little sprout was
already visible.
Figure 1.3: The effect to the growth of mung bean
watered by a 1.2% salinity of water; Day 1.

Figure 1.3 show no progress to the growth of the

mung bean watered by a 1.2% salinity of water
on its first day.

Figure 2.2: The effect to the growth of mung bean

watered by a 0.9% salinity of water; Day 2.
Figure 2.2 still show no progress to the growth
of the mung bean watered by a 0.9% salinity of
water on second day.

Figure 3.1: The effect to the growth of mung bean

watered by a 0.6% salinity of water; Day 3.

Figure 3.1 shows a little progress to the growth

of the mung bean watered by a 0.6% salinity of
water on its third day where the little sprout
Figure 2.3: The effect to the growth of mung bean
watered by a 1.2% salinity of water; Day 2. developed to be much longer from the second
day.
Figure 2.3 still show no progress to the growth
of the mung bean watered by a 1.2% salinity of
water on its second day.

Figure 3.2: The effect to the growth of mung bean

watered by a 0.9% salinity of water; Day 3.

Figure 3.2 also already show a progress by

Figure 3: The effect to the growth of mung bean
watered by a regular water; Day 3. managing to developed a little sprout to the
growth of the mung bean watered by a 0.9%
Figure 3 already show a progress to the growth salinity of water on its third day.
of the mung bean watered by regular water on its
third day where the bean developed a little
sprout.
Figure 3.3: The effect to the growth of mung bean
watered by a 1.2% salinity of water; Day 3.
Figure 3.3 still show no progress to the growth
of the mung bean watered by a 1.2% salinity of
water on its third day.
Figure 4: The effect to the growth of mung bean
watered by a regular water; Day 4.
Figure 4 already show a progress to the growth
of the mung bean watered by regular water on its
third day where the bean developed a much
longer sprout compared to the third day of
observation.
Figure 4.1: The effect to the growth of mung bean
watered by a 0.6% salinity of water; Day 3.
Figure 4.1 shows the most progress to the
growth of the mung bean watered by a 0.6%
salinity of water on its fourth day where the
sprout developed to be the most longer among
the four container of mung beans being watered
by a different salinity of water, from the third
day.
Figure 4.2: The effect to the growth of mung bean
watered by a 0.9% salinity of water; Day 3.
Figure 4.2 show a much progresive progress by
managing to developed a little sprout to the
growth of the mung bean watered by a 0.9%
salinity of water on its fourth day compared to
the third day.
Figure 4.3: The effect to the growth of mung bean
Figure 5.1: The effect to the growth of mung bean
watered by a 1.2% salinity of water; Day 3.
watered by a 0.6% salinity of water; Day 7.

Figure 4.3 still show no progress to the growth

of the mung bean watered by a 1.2% salinity of
water on its fourth day.
Figure 5: The effect to the growth of mung bean
watered by a regular water; Day 7.
Figure 5 shows show a progress to the growth of
the mung bean watered by regular water on its
third day where the bean developed a sprout
which is long and it also managed to crawl
outside the container, compared to the last figure
which is the fourth day of observation it
developed a long sprout or the stem, where a
two more days already passed; the fifth and sixth
day, that made it more possible to grow longer
spending a span of 7 days.
Figure 5.1 shows the most progress to the
growth of the mung bean watered by a 0.6%
salinity of water on its seventh day where the
sprout developed to be the most longest among
the four container of mung beans being watered
by a different salinity of water, where it has the
longest sprout and managed to developed a lot
of leaves compared to the other three container,
compared to the last figure 4.1 which is the
fourth day of observation, two more days
already which passed; fifth and sixth day, this
made the mung bean watered by 0.6% salinity of
water to grow aa a more healthy plant just in the
span of seven days.
Figure 5.2: The effect to the growth of mung bean
watered by a 0.9% salinity of water; Day 7.
Figure 5.2 show a much progressive progress
compared to the last figure which is the figure
4.2; the fourth day observation of the mung bean
watered by a 0.9% salinity of water, by
managing to developed a much longer sprout or
stem, but from the observation the sprout is
much thinner and not healthy at all, this growth
happened in the span of seven days.
Figure 5.3: The effect to the growth of mung bean
watered by a 1.2% salinity of water; Day 7.
Figure 5.3 still show progress to the growth of
the mung bean watered by a 1.2% salinity of
water on its seventh day where no sprout is
visible, and it didn’t managed to grow.
Salinity caused a significant reduction in plant
growth and associated developmental
parameters, since mung bean plant is very
sensitive to salt condition, selection of salinity
tolerant genotypes becomes important for mung
bean improvement. The objective of this study
was to evaluate the tolerance of 10 mung bean
genotypes pieces to salinity at seedling stage
under different levels. After observing the mung
bean for seven days, each day each container of
mung bean was changing but those change are
not similar from each other because of the level
of salinity of H2O to be used while watering the
beans.
At Day 1 there is no progress of growth in the
mung beans watered with water with a salinity
of 0.9% and 1.2% as well as the regular water
but the mung beans watered with 0.6 salinity
showed a progress by showing a little sprout,
when the Day 6-7 came, the difference between
the mung beans being watered by different
salinity of water was really visible to observed
among the four container of mung beans
watered by different level of salinity, the mung
beans watered with 0.6% of salinity show the
highest progress who happened to developed
the longest sprout among the for, while the
remaining three also managed to grow a sprout
and made a progress but not as healthy and
good as the mung bean with 0.6% of salinity.
This shows that the increasing salt
concentration significantly reduced the
percentage germination, fresh and dry weights,
protein and carbohydrate contents of all plant
parts, leaf area, shoot and root lengths, shoot/
root ratio, chlorophylls a, and b, and total
chlorophyll in both the cultivars.
The result showed that different salinity stress
levels affected all seedling characters, and each
genotype had different responses to salt stress.
There was a significant interaction effect
between mungbean genotypes and salinity
levels on speed of germination, vigor index,
germination, normal seedling fresh weight, nor-
mal seedling dry weight, and number of lateral
roots. The increasing salinity level was
significantly reduced the speed of germination.
The speed of germination is one of the pa-
rameters that indicate growing strength of seed
vigor which is more sensitive parameters than
germination rate. Speed of germination shows
the number of normal seedlings per day. High
speed of ger-mination reflects the vigor of
seeds, since the seeds can germinate rapidly in
a relatively short time.
The mungbean Vigna radiata exhibited
significant and high variations for adaptation to
salt stress. Salinity stress involves changes in
various physiological and metabolic processes,
varies with stress severity and its duration and
ultimately inhibits crop production. Initially, soil
salinity represses plant growth through osmotic
stress, which is then followed by ion toxicity.
Salinity stress causes a significant reduction in
mung bean yield through decline in seed
germination.
Conclusion
Temperature, sizes and amount of catalase affect
the enzyme's function and rate of reaction. First,
the temperature affected the enzyme's function
because as the temperature increases the rate of
reaction also increases, therefore the enzyme
was able to catalyze the reaction more rapidly.
Second, the size of particles, because smaller
particles consume less energy than larger ones to
break down molecules, therefore the reaction
would happen quickly. And last, the amount of
catalase because if the particles have a large
amount of catalase like liver, it lowers the
activation energy, so the rate of reaction will
speed up. Therefore, this experiment indicated
that catalase breaks down hydrogen peroxide
faster in liver cells than other cells such as
potato and carrots.
Mung beans are one of the more salt-tolerant
varieties of beans. This experiment discovered
the effects of salinity on mung beans and in
seven-day period, it was seen that salinization
had bad effects on mung bean growth. While, on
regular water, the mung bean growth was more
prolific. It is because the high salt content can
reduce germination rates and cause plants to
grow poorly and saltwater evaporates faster than
tap water which caused the deprivation of
moisture. To conclude, it shows that the higher
the salinity, the less growth of mung bean.
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Bender, M. (2014, September 29). Salinization
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https://www.scribd.com/doc/260450708/mung-
bean-lab
Kim, C. (2009, October 16). Relationship
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August 11, 2019, from
https://www.slideshare.net/christok77/biology-
catalase-lab

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