This Study Resource Was: Bromelain As An Enzyme

Bromelain as an Enzyme
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Biol_1031
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Friday 5:00-7:50 PM
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Title: Bromelain as an Enzyme
Abstract:
The purpose of this lab is to determine the effect that Bromelain has on Gelatin and what
happens in that reaction when you change the temperature and pH levels. Pineapples are
classified as bromeliads. Bromelain is a proteolytic enzyme that breaks down proteins into their
amino acids by hydrolysis (Pendarvis, Crawley, 2014). The effect of artificial and natural
bromelain had on the gelatin was determined. The effect that the water temperature had on the
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bromelain activity rate was also determined. Also, the effect of pH on the bromelain activity rate
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was also determined. The bromelain enzyme effected the gelatin the most at room temperature.
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The optimum pH level in this activity was 13.
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Introduction:
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For most chemical reactions in cells to proceed at a rapid pace, a catalyst is needed. A
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catalyst is an agent that speeds up the rate of a chemical reaction without being permanently
changed or consumed by it. In living cells, the most common catalysts are enzymes, which are
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proteins (Brooker, 2017). Enzymatic reactions occur in every cell. It is important to understand
and see how different enzymes may affect specific solutions. In each of these reactions, there is
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an enzyme molecule and a substrate. The substrate has a specific shape and function as well as
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the enzyme it is because of this that the chemical reaction can occur.
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Every time an enzyme and substrate form a bond, a chemical reaction occurs. However,
there are different conditions which can affect the final products of these chemical reactions.
Although the enzyme is the catalyst to every chemical reaction, the enzyme may be affected by it
surrounding conditions. These conditions can also help to either slow down or speed up the
chemical reactions. In many studies some of the most prominent conditions that I have come
across are the temperature and pH levels. In many cases the temperature can either speed up or
slow down the reaction, the same goes for the pH levels. The fascinating thing about all of this is
with each different enzyme-substrate bond, there is a different pH and temperature level which is
optimum for it.
Material Methods
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Materials:
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Test tube rack, test tube clamp, eye protection, test tubes, syringes, water, fresh pineapple
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juice, canned pineapple juice, gelatin, ice container, hot plate, ice, pipettes, HCI, NaOH, pH 3,
pH 5, pH 7, and pH 11.
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Methods:
This study was conducted in a biology lab on Baton Rouge Community College Mid-City
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campus. There were 3 procedures in this lab, in which the affect that bromelain had on gelatin
was studied by itself and then with manipulated temperature and pH levels. Each lab group in the
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class studied the different reactions and then compiled all the information together.
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Determination of bromelain effect on gelatin (procedure 1)

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We used three separate test tubes labeled “water”, “fresh”, and “canned”. We added 3 ml
of warm gelatin to each tube. With separate syringes we added 2 ml of water to the test tube
labeled “water”, 2 ml of fresh pineapple juice to the test tube labeled “fresh”, and 2 ml of canned
pineapple juice to the test tube labeled “canned”. We then placed all the test tubes on ice, let it sit
for 5-10 minutes, then observed and recorded the reactions in each tube.
Determination of effect of temperature on bromelain activity rate (procedure 2)
We used three separate test tubes labeled “water”, “hot”, and “cold”. We added 2 ml of
water to the test tube labeled “water” and placed it in the test tube rack. Then added 2 ml of fresh
pineapple juice to the test tubes labeled “cold” and “hot”. Leaving the test tubes labeled “water”
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and “cold” at room temperature. We placed the test tube labeled “hot” into a water bath for five
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minutes with the temperature at 70 degrees Celsius. After the five minutes was up, we removed
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the test tube labeled “hot” from the water bath, placing it back in the test tube rack with the
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others. We added 3 ml of warm gelatin to each test tube. Took all three test tubes and submerged
them in ice, let it sit for 5-10 minutes, then observed and recorded the reactions.
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Determination of effect of pH on bromelain activity rate (procedure 3)
We used a total of ten test tubes and labeled them “1” through “10”. With a pipette we
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added the following to the test tubes: test tube “1”: 2 ml of water; test tube “2”: 2 ml of fresh
pineapple juice; test tube “3”: 1 ml of HCl and 1 ml of fresh pineapple juice; test tube “4”: 1 ml
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of HCl and 1 ml of water; test tube “5”: 1 ml NaOH and 1 ml of fresh pineapple juice; test tube
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“6”: 1 ml of NaOH and 1 ml of water; test tube “7”: 1 ml of pH 3 and 1 ml of fresh pineapple
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juice; test tube “8”: 1 ml of pH 5 and 1 ml of fresh pineapple juice; test tube “9”: 1 ml pH 7 and
1 ml of fresh pineapple juice; test tube “10”: 1 ml of pH 11 and 1 ml of fresh pineapple juice. We
then mixed components of each test tubes by swirling the contents, then let them sit for three
minutes. After the three minutes was complete we added 3 ml of gelatin to each test tube. We
then placed all 10 test tubes in ice and let it sit for 5-10 minutes. After the time was up we
observed and recorded all reactions.
Results
In the bromelain activity we observed the chemical reaction with the enzyme bromelain
and gelatin. We did this with fresh and canned pineapple juice, while also manipulating
temperatures as well as pH levels. The bromelain from the canned pineapple juice and water
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mixture from procedure one had solidified into a gel. In the second procedure the plain water
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without any temperature manipulation solidified best. In procedure three none of the bromelain
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test tubes solidified, but the water solidified best with the pH of 13 (NaOH).
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Discussion
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In the first procedure we only tested the affects of bromelain by itself mixed with gelatin.
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Then a test tube of just water and gelatin. What we found during this procedure was that the tube
with only water and gelatin formed a gel, as well as the canned pineapple juice mixed with
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gelatin. The fresh pineapple juice mixed with gelatin remained a liquid when we pulled it out of
the ice.
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During procedure two, we manipulated the temperature of the water and added only fresh
pineapple juice. What we found to be true with this experiment, was that the two test tubes that
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had the fresh pineapple juice and manipulated temperature changes, no chemical reaction took
place and the solution remained liquid. The plain room temperature water with no bromelain in
the mixture was the only test tube which solidified into a gel.
During procedure three, we manipulated the pH levels with in the test tubes. Some test
tubes had a manipulated pH with the bromelain, others had a manipulated pH level with no
bromelain, just water. One test tube was just water. Similarly, to procedure two, we found that all
test tubes with bromelain in the mixtures had no chemical reaction within the time we were given
and remained liquid. However, the plain water test tube did solidify likewise to the other
procedures. Also, the water with the pH level 13 (NaOH) was solidified in the time frame that we
had to conduct this experiment.
Conclusion
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Through our experiment the reaction between the enzyme bromelain and gelatin was
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tested. Using gelatin as our control, we changed the type of bromelain used, as well as
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manipulated the temperature and pH levels of the different solutions in the test tubes to see
which would solidify into a gel. We concluded that using the bromelain from canned pineapple
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juice worked best for the chemical reaction to occur that solidifies the solution. We also found
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that manipulating the temperature and pH levels of the fresh pineapple juice had no affect on the
chemical reaction to make it solidify.

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Acknowledgements
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I would like to thank my group partners, Fallon Daniels, and Myra Jackson for their help during
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this experiment.
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References
Pendarvis, M. & Crawley, J. (2014) Exploring Biology in the Laboratory (Second Edition).
Englewood: Morton Publishing Company.
Brooker, R. & Widmaier, E. & Graham, L. & Stiling, P. (2017) Biology (Fourth Edition). New
York: McGraw-Hill Education.
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Bromelain as an Enzyme

optimum for it.

Determination of bromelain effect on gelatin (procedure 1)

Determination of effect of temperature on bromelain activity rate (procedure 2)

Determination of effect of pH on bromelain activity rate (procedure 3)

observed and recorded all reactions.

had to conduct this experiment.

chemical reaction to make it solidify.

Englewood: Morton Publishing Company.

York: McGraw-Hill Education.

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