Experiment 7

Yeast Respiration

Objective

1) To understand the basic concept of cellular respiration

Introduction

Yeast (saccharomyces) is a common single-celled fungus. They are microscopic plants

that exist naturally on Earth. They are known for their ability to ferment the carbohydrates in the
flour and gain energy from this digestion. Some food products created by yeast include bread,
beer, wine and cheese. Yeast is mistaken as lifeless, dry powder used to make bread, but, it
becomes active in moist environments. Yeasts secrete digestive enzymes outside their cells and
absorb digestion products back in. This process is called extracellular digestion. Yeasts are
considered to be heterotrophs, meaning that they get food from other organisms. They have a
special molecule in their cell wall called chitin in their cell wall that allows them to do so.

In cellular respiration, energy in food molecules is released and converted to a form that
can be used by the cell, which is ATP. The term respiration is often used to mean breathing or
simply the process of inhaling and exhaling. For clarity, the term cellular respiration is used to
refer specifically to a series of enzymatic reactions that can occur in the presence or absence of
oxygen and make energy available to the cell. Some cells use anaerobic respiration to produce
ATP, a form of chemical energy. Anaerobic respiration means that there is no use of oxygen
required for this to take place. Cell respiration refers to many different ways that can metabolize
glucose, a monosaccharide. All those different ways start off with Glycolysis, it breaks down a
glucose molecule into 2 pyruvate molecules (3-carbon molecule). In this process, there is a gain
of 2 ATP molecules. The breakdown of organic material for ATP production is called
fermentation
 The changing temperature can have effects on many living things. Enzyme-catalysed
reactions are very sensitive to small changes in temperature. The metabolism of some organisms
can depend on their internal body temperature, which is determined by its environment and often
by the surrounding temperature. Yeast is one of those organisms. It is used to leaven bread (make
it rise). Yeast does this by fermenting sugar, producing carbon dioxide as a waste product which
is lost. However, some of the carbon dioxide is trapped which makes the bread light, small air
pockets is the reason behind this. If the yeast is not warmed properly, it will not be of much use
as a leavening agent, the yeast cells will burn sugar much too slowly.

Methodology

1) 2 test tube was prepared and labelled as 1 and 2.

2) 1 ml of yeast solution was added inside each test tube which in test tube 1 contained 1 ml
of boiled solution and test tube 2 consist of pre-boiled yeast.
3) 4 ml of glucose was added in each test tube together with 200 l of phenol red.
4) Both tube was pre incubated for 60 minutes and being observe for any changes of
appearance such bubble formed and color changes.

Result
Figure 1: Yeast Solution at zero minutes.

Figure 2: Results of Yeast Solution at 20 minutes and 1 hour after.

Table 1: Results obtained after 1 hour incubation.

 Yeast Sample Bubble Color

Boiled No bubble form Orange

Unboiled Bubble form Yellow

Discussion

Results in this lab yielded distinct conclusions regarding the ability for yeast to
metabolize simple and complex sugars. Because the organism uses enzymes to initiate the sugars
into glycolysis, it makes sense that the enzymes are able to catalyze specific sugars based from
this, the main objective of this experiment was to understand the basic concept of cellular
respiration. Cellular respiration is spontaneous chemical process wherein a cell makes energy by
the transformation of energy molecules such as sugars and fatty acids into ATP and certain by-
products. Cellular respiration is divided mainly into aerobic respiration and anaerobic
respiration.

As shown in figure 1, 2 test tube was prepared which contained a yeast that was boiled
and preboiled and after 1 hour, the results obtained showed bubbles presence in the test tube with
preboiled yeast compared to the second test tube as noted in table 1. Based form the result, its
shows the cellular respiration occur in the test tube 1 (preboiled yeast). This was because in
theory, yeast will use the glucose inside as a food source. When the cells of yeast used the sugar,
it will gets the energy and produce the carbon dioxide (CO2) also the water (H2O). This means,
the presence of glucose in the solution, enhances the cells of the yeast breaks the sugar to gets
the energy, releasing the carbon dioxide and water, this bubbles presence can be seen in the
figure 2, the bubbles measured was 3 cm.

To be more specific, in cellular respiration the glucose molecules presence in the solution
was transformed into pyruvic acid in the cytosol by using 2 units of ATP. In this process, 4 units
of ATP was produced and the net total was 2. The pyruvic acid produced will transformed into
Acetyl CoA and this product will goes to the next process called TCA Cycle, which the product
will goes to the numerous transformation and results into 8 units of NADH, 2 units of FADH and
4 units of ATP. The total units of ATP produced was 36 units. This means, yeast was a facultative
anaerobe, it capable of making ATP by aerobic respiration when oxygen is available. The amount
of carbon dioxide gas produced reflects the rate of cell respiration because CO2 is a byproduct of
anaerobic cell respiration in yeast. So an increase in carbon dioxide production means an
increase in the rate of cellular respiration. Glucose are simpler carbohydrates which can speed up
the rate of cellular respiration more that the complex carbohydrates. If the amount of substrate is
increased then the rate of cellular respiration and carbon dioxide production will also increase.
This is because an increase in the availability of the substrate will allow more yeast cells to use
the substrate for cellular respiration in the mitochondria and the more yeast cells working on
cellular respiration at a given time, the more ATP and CO2 will produced.

But, for the boiled yeast tested in glucose, there was no presence of bubbles in the test
tube as shown in the figure 2. This was because, by using the high temperature will contribute to
molecular energy for the yeast cells. If high temperature were used, denaturing would occur in
the yeast cells, killing the yeast and because of this, the yeast were not be able to proceed with
cellular respiration. Since there was no cellular respiration occur, there was no CO2 and H20
produce during the reaction occur thats why the bubbles was not presence in the test tube. From
this reason, the cellular process and TCA cycle was not being produce in this reaction.

Phenol red exists as a red crystal that is stable in air. This solution contains a pH indicator
which helps in monitoring of the pH changes in the cell culture, its color exhibits a gradual
transition from yellow to red over the pH range 6.8 to 8.2. As seen in figure 1 and 2, the color of
test tube 1 which contained preboiled yeast changes to yellow color while for the second tube the
color changes to orange color. By using the pH indicator will indicates the CO2 is higher or
lower in the dilution. Based from the results indicates that high CO2 concentrations in water will
becomes acidic and at low CO2 water becomes more basic. Phenol red is yellow under acidic
conditions was at high CO2 concentration, red under basic or alkaline conditions at low CO2 ion
concentration and orange under neutral conditions. This indicates that from the result obtained,
test tube 1 shows the dilution was in acidic condition which high with CO2 concentration, and
the pH was below 6.8 while the other test tube contained of yeast that already die due to the high
temperature, therefore there was no CO2 being produced and because of this the color changes to
orange color indicates the dilution was in the neutral condition.

Conclusion
 To conclude, the objective of the experiment was achieved. The rate of cellular
respiration was affected by the condition in which it take placed. When a yeast and a simple
sugar (glucose) was presence during the cellular respiration, the production of CO2 will formed
more while the process of CO2 will be reduced since the yeast was die in high temperature
(boiled), thats why the yeast was unable to do the cellular respiration. While by using the pH
indicator indicates that the higher the CO2 concentration in the solution, the more acidic were be
in water condition.

Reference

Delos Reyes (2006), Introduction to Biology: Principles and Processes (6th ed). Department of
Biology. [Retrieved 29th Nov 2015]

Campbell, N.A, Reece, J.B & Meyers (2006). Biology Pearson Education.

Shimomura-Shimizu, M. (2009, October 18). Yeast Based Sensors. National Center for
Biotechnology Information. At: http://www.ncbi.nlm.nih.gov/pubmed/20087724. [Retrieved 29th
Nov 2015]

Day, Sam Adam (2013), "Factors Affecting Enzyme Activity." A Level notes. At:
http://alevelnotes.com/Factors-affecting-Enzyme-Activity/146. [Retrieved 29th Nov 2015]