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DEPARTMENT OF PURE AND APPLIED CHEMISTRY

Visayas State University, Baybay, Leyte


CHEM 31 Biochemistry
Laboratory Report

Name: Loise Mariel C. Bibera Date Performed:

Lab. Schedule: M-F 7:00-10:00 Date Submitted: 06/03/15

Group No: 4 Score:

Experiment No. 12

Energy Production in the Living Cell

I. Objective:
1) Observe the reaction of glucose in glycolysis and fermentation.

II.Results
A. Fermentation
Test Tubes Solution Rank
1 10 mL yeast suspension filled 4
with water
2 10 mL yeast suspension filled 2
with glucose solution
3 10 mL yeast suspension filled 3
with starch solution
4 10 mL boiled yeast suspension
filled with glucose solution
5 10 mL yeast suspension filled 1
with sucrose solution

B. Glycolysis
Time (min) Observations
Test Tube
1 2 3
(Water + Yeast (Glucose + Yeast (Glucose + Boiled
Solution) Solution) Yeast Solution)
0 Blue Blue
20 Blue Deep Blue Green
(bottom)
40 Deep Blue Green Deep Blue Green (all)
(bottom)
60 Light Blue Green Light Blue Green
(bottom)
80 Light Blue Green Light Blue Green
(Bottom)
120 Light Blue Colorless

III. Discussion

In order to keep cells functioning they require a constant supply of energy to generate and
maintain their biological order. This energy in the form of ATP (Adenosine triphosphate) is
derived from the chemical bond energy in food molecules, which thereby serve as fuel for cells.
These functions include replacement of cell parts, growth and cell division, and special functions
(such as secretion, absorption, contraction, or signaling).

Sugars are particularly important fuel molecules, and they are oxidized in small steps to carbon
dioxide (CO2) and water. In this part we trace the major steps in the breakdown, or catabolism,
of sugars and show how they produce ATP, NADH, and other activated carrier molecules in
animal cells. In the laboratory procedure the class had concentrated on glucose breakdown,
since it dominates energy production in most animal cells and will tackle the two main catabolic
reactions, glycolysis and fermentation.

A. Fermentation

Fermentation is a process adopted, typically, by anaerobic organisms to obtain ATP without the
use of oxygen. Saccharomyces cerevisiae, or bakers yeast, is a unicellular fungus that uses
both fermentation and respiration when needed. When yeast is in the presence of oxygen it
performs cellular respiration, but when oxygen is absent it undergoes alcohol fermentation. In
alcohol fermentation, the sugar is converted intotwo 3-Carbon sugars known as pyruvate. The
pyruvate is then converted intoethanol alcohol in two steps. In the first step, it is converted into
acetaldehyde with the release of carbon dioxide (CO2), the independent variable of the
experiment. Next the acetaldehyde is reduced to ethanol. In this lab alcohol fermentation of
conducted.

The fermentation procedure required 5 test tubes with 5 varying mixtures of yeast suspension
and carbohydrates such as glucose, starch and sucrose having one test tube as control (which
consist of yeast suspension and water. The fourth mixture which consisted of a boiled yeast
suspension filled with glucose was not conducted due to the inadequate time to bring the yeast
suspension to a boil. After the preparation of the indicated mixtures the test samples were
heated in a water bath with a constant temperature of 37oC. The summary of results was
tabulated in the first part of this laboratory report. Based on the table the test tube sample that
provided the fastest rate of CO2 formation was test tube 5 containing yeast and sucrose
followed by test tube 2 (yeast + glucose) third was test tube 3 (yeast + starch) and lastly the
control (yeast + water).

Upon examining the structures of each of the sugars it suggests that the test tube
containing glucose would ferment the most because it would require less work to break down
into pyruvate and acetaldehyde. Sucrose would evolve the next highest amount of CO2,
because its structure was a bit more complex, being a disaccharide (two sugars) made up of
glucose and fructose. I seemed that it may take more energy to break down the sugar.
Considering the fact that a 6-Carbon sugar attached to a 5-Carbonsugar would limit production
of CO 2 . The third in rank would be starch since it is a long chain of polysaccharide making it
even harder to break. However, there is a discrepancy in the results of glucose and sucrose.
Instead of glucose being the fastest it showed that sucrose fermented first. It is possible that
yeast has enough enzymes to break down the sucrose, allowing for rapid fermentation of the
sugar. The result with the control (yeast suspension + water) was expected since it does not
contain any carbohydrate to ferment with.

B. Glycolysis

In this exercise you will observe fermentation in yeast cells. Oxidation of glucose
during this process yields hydrogens which must be "picked up" by hydrogen
acceptors. Methylene Blue (oxidized) will accept hydrogens to become Methylene
Blue (reduced). The oxidized form of methylene blue has a definite blue color,
whereas the reduced form is colorless.

Cellular respiration is a process that most living organisms undergo to createand obtain
chemical energy in the form of adenosine triphosphate (ATP). Theenergy is synthesized in three
separate stages of cellular respiration: glycolysis,citric acid cycle, and the electron transport
chain. Glycolysis and the citric acid cycleare both anaerobic pathways because they do not need
oxygen to form energy. Theelectron transport chain however, is aerobic due to its use of
oxidativephosphorylation. Oxidative phosphorylation is the process in which ATP moleculesare
produced with the assistance of oxygen molecules. (Campbell et al 2008)
IV. Conclusion

Cells need energy to accomplish the tasks of life. Beginning with energy
sources obtained from their environment in the form of sunlight and organic
food molecules, eukaryotic cells make energy-rich molecules like ATP and
NADH via energy pathways including photosynthesis, glycolysis, the citric acid
cycle, and oxidative phosphorylation. Any excess energy is then stored in
larger, energy-rich molecules such as polysaccharides (starch and glycogen)
and lipids.
V. Answers to Questions

C6H12O6 2 Ethanol + 2 CO2 + Energy (2 ATP + Heat)

VI. References

Energy Production In A Cell

http://www2.sunysuffolk.edu/czuraa/BIO130LectureMaterials/BIO130Chapter25Handout.pdf

How Cells Obtain Energy from Food


http://www.ncbi.nlm.nih.gov/books/NBK26882/

FERMENTATION OF VARIOUS SUGARS IN


BAKERS YEAST
http://www.academia.edu/5434410/Fermentation_Formal_Lab_Report

Alcoholic Fermentation in Yeast

http://www.instruction.greenriver.edu/kmarr/biology%20211/Labs%20and%20ALEs/B211%20L
abs/B211%20Labs/5%20_Lab%205_Alc%20Ferm%20in%20Yeast_F2009.pdf