UNIVERSITY OF SANTO TOMAS

BOTANY
1G-PH
Anaerobic Respiration: Alcoholic Fermentation
Members: Arrojo, Shiela Marie; Bascuguin, Ira Sheinel;
Uy, Roi Marion
I.

Introduction
The single- celled fungus Sacchorymes cerevisiae, a yeast, is capable of both
aerobic and anaerobic when oxygen is present it breaks down glucose anaerobically
following the metabolic sequence of glycolysis, intermediate reactions, krebs cycle,
and electron transport system. Without oxygen, only glycolysis takes place. Glucose
is metabolized to pyruvate which in turn, is converted to two waste products. Carbon
dioxide (CO2) and Ethanol.

II.

Objectives
1. To prove that yeast is capable of aerobic and anaerobic respiration.
2. To prove that ethanol and carbon dioxide is present in anaerobic respiration
by means of phenol red solution (pH indicator) and iodoform test.
3. To prove that yeast undergone alcoholic fermentation.

III.

Materials
3 fermentation tubes/
Erlenmeyer flasks
Cotton
Beaker
Graduated cylinder
Weighing scale
Pasteur pipette
Wire loop
3 test tubes
Test tube rack
Nutrient medium:
a.
10% glucose
b.
2% peptone
c.
0.5 yeast extract
Distilled H2O
Phenol red
1.25 ethanol
Strong I2K
1.5 M NaOH
Methodology
-Flowchart Form

IV.

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-More detailed:
1. Fill 3 Erlenmeyer flasks with nutrient medium, it is composed of 50 ml 10%
glucose, 30 ml 10% peptone and 0.5% yeast extract.
2. Plug the mouth of Erlenmeyer flask with cotton and sterilize for 1 hour to
destroy any foreign microorganisms. Cool the set up before proceeding to the
next step.
3. Erlenmeyer flask A serves as the control substance and demonstrates the
effect of sterilization.
4. Add live yeast suspension (1 mongo grain) to Erlenmeyer flask B. Observe
the result after 2 days. The microorganism had undergone aerobic respiration
using the dissolved oxygen to build up its cell population. The CO 2 of aerobic
respiration combined with water formed a weak acid Carbonic acid.

(CO2+ H2O
H2CO3)
5. Add live yeast to flask C and observe the results for 5 days. The organism
utilized the excess glucose anaerobically and produced CO 2 and ethanol.
Remove the cotton plug and smell the content.
6. To prove the presence of CO 2, prepare and number two test tubes in a test
tube rack. With a disposable Pasteur pipette, transfer to tube 1 a pipetteful of
fermentation medium from flask C. TO tube two, place a similar pipetteful of
distilled water. And to both tubes add one drop of phenol red solution. Phenol red
indicator is a pH indicator.
7. To prove the presence of ethanol, you will do iodoform test.
8. Add to each tube 1 ml strong I2KI and 1.5 M NaOH. Mix the content of each
tube and allow to stand for 5-10 mins. If ethanol is present, it will react with iodine
in the presence of NaOH to form iodoform, which will settle out as yellow
precipitate.
9. Examine the demonstration pure culture of the yeast on a solid medium. To
see the yeast, mix the remaining medium in fermentation tube C, and with a wire
loop, transfer a loopful of it in a glass slide and place a cover slip.
10. Examine under LPO and HPO of a microscope. Make an enlarge sketch of a
few yeast cells.
V.

Results and Explanations

Erlenmeyer Flasks
A (Control Set Up)
B (Two days after)
C (Five days after)

Appearance
More yeast population
through O2

Product Formed
CO2+ H2O
H2CO3
Carbonic Acid

Proving the presence of CO2:

Test Tubes
1 (Fermentation Medium)
2 (Distilled H2O)

Appearance
Yellow color
Red color

Product Formed
acidic
basic

Iodoform Test (Proving the presence of ethanol):

Test Tubes
1 (2.5ml distilled H2O)
2 (1.25ml distilled H2O and 1.25% ethanol)
3 (2.5ml from fermentation medium flask C)

Iodoform Result
(+/-)
+
+

After two 2 days, we had observed that E. Flask B has more yeast cell population.
Therefore we conclude that E. Flask B performed an aerobic respiration. Where it
used up the dissolved oxygen from the CO 2 it produced. The carbon dioxide of
aerobic respiration from the yeast combined with water produces a weak acid which
is Carbonic acid. Based on our observation the yeast on Flask C after five (5) days,
it performed anaerobic respiration. When the cotton was plugged off on the mouth E.
flask and the content smells like alcohol. Which proves that E.Flask C. undergone
anaerobic respiration; the organism utilized the excess glucose anaerobically and
produced CO2 and ethanol (alcohol). Since there is no oxygen in the respiration it
became alcoholic fermentation. E.Flask C produced CO2 and ethanol as waste
products and is evident when smelling the said flask. When we added a drop of
phenol red solution to tube 1 (fermentation medium) and tube 2 (distilled H 2O), we
found out that tube 1 appears red therefore it is acidic then, test tube 2 appeared
yellow therefore it is basic. The iodoform test that we did proved the presence of
ethanol. Among the 3 tubes, test tube 2 which contained also 1.25% ethanol and
test tube 3 which contained 2.5 mL medium from E.flask C came out positive
because the ethanol reacts with iodine in the presence of NaOH forming an iodoform,
which can be seen as the yellow precipitate settled out when we add 1 mL of strong
I2KI and 1.5 mL 1.5 M of NaOH in the tube.
VI. Conclusion
The yeast has formed oxygen which mixed with water and formed weak acidCarbonic Acid (CO2+ H2O
H 2CO3) proven by the test with the phenol red
indicator. On the other hand, through the iodoform test, the yeast has been found to

produce ethanol which was indicated by the presence of the iodoform (yellow
precipitate).
Therefore, the single-celled fungus Saccharomyces cerevisiae, the yeast used
for this experiment had indeed undergone Aerobic through the use of oxygen,
Anaerobic respiration producing CO2 and Alcoholic Fermentation by the presence of
ethanol in the yeast.
VII. References
http://www.anaerobicrespiration.net/general/anaerobic-respiration-in-yeast/
http://www.passmyexams.co.uk/GCSE/biology/aerobic-and-anaerobicrespiration.html