Influence of pH on the function of Yeast

Unit 6 Coursework
Irukshi Anuprabha Silva
01/04/2015

Influence of pH on the function of yeast Research & Rationale

Abstract
The aim of this experiment is to investigate the influence of varying pH from pH 1 to 10 on the
functioning of Saccharomyces cerevisiae in order to find the optimum pH for an optimum rate of
reaction. Knowing the optimum pH of yeast would be beneficial for various industries like the
food industry.
From the results, the optimum pH was found to be at pH 4 where the mean rate of reaction was
the fastest recorded at 0.033cm3/sec. The results showed that both a positive and negative
correlation is present. However, the negative correlation is more dominant hence, the overall
correlation is negative between pH and rate of respiration in yeast.
Null Hypothesis
There is no correlation present between pH and the rate of respiration in yeast.
Alternate Hypothesis
A correlation is present between pH and the rate of respiration in yeast.
Research and Rationale
This experiment aims to investigate the effect of pH on the rate of respiration of yeast and how
this could be beneficial to the food industry.
According to Source 2 in the case of brewing beer, in most cases the brewer does not have to
worry about pH - this is because, if properly done, the brewing processes tend to settle at an
adequate pH. As a result pH is one of the last frontiers in brewing practice that home brewers
will encounter and worry about.
But at the same time, according to Source 3, Yeasts play a prominent role in wine
fermentations, which can strongly affect the quality and flavour of the final product (Querol and
Fleet, 2006).
Saccharomyces cerevisiae [also known as bakers yeast] is a species of yeast that is commonly
used to raise dough when baking bread or for the fermentation of beer or wine.
S. cerevisiae is a fungus which contains multiple useful enzymes that are required in the process
of fermentation. Enzymes are proteins which are made of amino acids. [Source 1] pH can have
an effect of the state of ionization of acidic or basic amino acids. If the state of ionization of
amino acids in a protein is altered then the ionic bonds that help to determine the 3-D shape of
the protein can be altered. This can lead to altered protein recognition and the enzyme may
become inactive. Changes in pH may not only affect the shape of an enzyme but it may also
change the shape or charge properties of the substrate so that either the substrate cannot bind to
the active site or it cannot undergo catalysis. Enzymes have an optimum pH, but this pH is not
common for all enzymes.
Hence, the aim of my experiment is to find the overall optimum pH for the respiration of yeast.
Word Count - 456
Sources:
Source 1: http://academic.brooklyn.cuny.edu/ - Date accessed 19/01/2015
Source 2: http://www.braukaiser.com Date accessed- 20/01/2015
Source 3: International Journal of Food Microbiology by F. No Arroyo-Lpez, Sandi Orli,
Amparo Querol, Eladio Barrio - Date accessed- 20/01/2015
Source 4: http://www.thebakerynetwork.com/ - Date accessed 16/01/2015
1

Source 5: Yeast Physiology and Biotechnology by Graeme M. Walker Date accessed

16/01/2015
Source 6: Use of Yeast Biomass in Food Production by Anna Halasz, Radomir Lasztity Date
accessed 16/01/2015
Influence of pH on the function of yeast Planning
Preliminary experiments were carried out to find a suitable ratio of the concentration of yeast to
the concentration of sucrose so that the volume of CO2 produced is within the range of the gas
syringe used. The experiment was carried out by activating the yeast solution in a conical flask
for 5 minutes in a water bath of 35oC. Next, the sucrose solution was added into the conical flask
and the volume of CO2 gas produced was recorded in different intervals of time. In this
experiment, pH will be the independent variable and the volume of CO2 produced will be the
dependent variable.
A Spearman rank correlation test will be used to analyze the data, which will be used to verify
the hypothesis.
Preliminary Results
Preliminary 1
Concentration
Time[min]

0
1
2
3
4
5
6
7
8

0.7M
Sucrose
(8% yeast)
Vol. of
CO2[cm3]

0.0
7.0
17.0
31.0
44.0
58.0
70.0
83.0
95.0

Preliminary 2
Concentration
Time[sec]

0.01M
Sucrose
(8% yeast)
Vol. of
CO2[cm3]

0
30
60
90
120
150
180

Hence, I conducted the next few preliminary

trials with a sucrose concentration of 0.1M in
an 8% yeast solution this concentration was
neither too high nor too low. I also increased
the activation time from 5 minutes to 10
minutes because at 5 minutes, the yeast
solution was not frothy, indicating that it had
not fully activated yet. I also decided to take
readings every 30 seconds this interval was
suitable as it gave me sufficient time to check
the temperature and maintain the water bath
at 35oC.
Word Count: 327

0.0
4.0
4.0
4.0
4.5
4.5
5.0

The first 2 preliminary trials were used to

find a suitable ratio of the concentration of
sucrose to the concentration of yeast. The
results indicate that a concentration of
sucrose that is neither too high nor too
low would be suitable as with a
concentration of 0.01M, the rate was too
slow while with a concentration of 0.7M,
the rate of reaction was too rapid and
exceeded the range of the 100cm3 syringe.

Preliminary 3 - Control
Time[sec]

0
30
60
90
120
150
180
210
240
270
300

Trial 1

Trial 2

Vol. of
Vol. of
3
CO2[cm ] CO2[cm3]
0.0
0.0
3.5
1.5
4.5
2.5
5.5
3.5
7.0
4.5
8.0
5.5
9.0
6.5
10.0
7.0
10.5
8.0
11.0
9.0
11.5
10.0

Mean
Vol. of
CO2[cm3]

0.00
2.50
3.50
4.50
5.75
6.75
7.75
8.50
9.25
10.00
10.75
2

Time
[sec]
0
30
60
90
120
150
180
210
240
270
300

Preliminary 4 - pH 1
Trial 1 Trial 2
Mean
Vol. of Vol. of
Vol. of
CO2
CO2
3
CO
2[cm ]
[cm3]
[cm3]
0.0
0.0
0.00
3.5
1.5
2.50
4.5
3.5
4.00
5.0
4.0
4.50
5.0
4.0
4.50
5.5
4.0
4.75
6.0
4.0
5.00
6.0
4.5
5.25
6.0
5.0
5.50
6.0
5.0
5.50
6.0
6.0
6.00

Preliminary 5 - pH 10
Trial 1 Trial 2
Mean
Vol. of Vol. of Vol. of
Time[sec]
CO2
CO2
CO2
3
3
[cm3]
[cm ]
[cm ]
0
0.0
0.0
0.00
30
2.5
3.0
2.75
60
2.5
4.5
3.50
90
2.5
5.0
3.75
120
2.5
5.0
3.75
150
3.0
5.0
4.00
180
3.0
5.0
4.00
210
3.0
5.0
4.00
240
3.0
5.0
4.00
270
3.0
5.0
4.00
300
3.0
5.0
4.00

The trials from preliminary experiments 3,4 and 5 were conducted in order to test my hypothesis
and to observe if any correlation is present. The buffer was added to the yeast solution before
activation and the volume of buffer added was equivalent to the total volume of yeast and
sucrose combined.
A trial experiment was carried out to find a suitable ratio for the concentration of sucrose to the
concentration of yeast as well as to test whether the hypothesis is valid or not. Trial experiments
1 and 2 show that the ratio of sucrose to yeast must neither be too high nor too low. If the
concentration was too high, it exceeded the capable range of the gas syringe and if the
concentration was too low, the rate at which CO2 was produced was too slow. From these two
trials, I decided to take a concentration of 0.1M sucrose and also decided to set a fixed time
period of 5 minutes 300 seconds. Next, trials 3, 4 and 5, the buffer solutions [which controlled
pH] were added along with the yeast solution for activation. This proved effective as with
different pH, the rate of reaction of yeast had changed. Moreover, for the main experiment, the
activation time was increased from 5 minutes to 10 minutes as at 5 minutes, no froth was
observed this could mean that the yeast had not been activated properly.
From the results obtained from preliminary experiments 3, 4 and 5, I confirmed that my method
and hypothesis is valid and carried out further investigations.
Word Count: 316

Final Method:
For the main experiment, the yeast solutions
along with the buffer solution were activated in a
water bath of 35oC at 10 minutes. Next, the
sucrose solution was added to the conical flask
and the volume of CO2 produced was recorded
for 5 minutes at intervals of every 30 seconds.
The conical flask was swirled at a constant,
moderate pace after the sucrose was added so that
the reaction would occur quickly or else, in a
time period of 5 minutes, if the conical flask had
not been swirled, the rate of reaction would have
Figure 1 A picture of the apparatus used in my
been too slow to give significant results. The gas
investigation
syringe was the most appropriate apparatus to
collect the volume of gas produced, however each
division represented only 1cm3 - so in the case of results that lie in between two divisions, there
was less precision in recording results which means that inaccurate results could have been
recorded. On the other hand, when measuring the volume of pH or distilled water, the measuring
cylinders used had appropriate divisions and had a high amount of precision for accurate
measurements to be made.
In order to maintain validity, the yeast used was from the same brand and taken from the same
container, the same conical flask and gas syringe were used. Moreover, the concentration of both
sucrose and yeast remained constant as the same volume of distilled water was used for all trials,
the same measuring cylinders were used all throughout and the lower meniscus was taken when
measuring volume eliminating the possibility of errors in measuring. The temperature was kept
constant by maintaining the temperature of the water bath at 35oC.
In order to maintain reliability, 3 trials were conducted for each pH and the mean volume of CO2
produced was calculated.

Risk Assessment:
For pH 1 and pH 2, the buffer solutions were prepared by mixing 0.1M hydrochloric acid with
0.1M of potassium chloride. Hydrochloric acid is a corrosive acid which can cause severe burns
of the skin and eyes hence rubber gloves and safety goggles were worn when handling the acid.
All glass apparatus were handled with care as when broken, pieces of glass can cause cuts and
wounds. The thermometer was handled with extra care because if broken, liquid mercury would
have leaked out and can cause mercury poisoning. In addition to these precautions, a lab coat
was worn at all times to avoid possible spillages.

Word Count: 415

Influence of pH on the function of yeast Observing and recording

Results:
0

30

60

90

120

150

180

210

240

270

300

Mean
ROR[cm3/sec]

0.0
0.0
0.0
0.00

0.5
0.5
1.5
0.83

1.5
2.0
3.0
2.17

2.5
3.0
4.0
3.17

4.0
4.0
5.5
4.50

5.5
5.5
6.5
5.83

6.5
6.5
7.5
6.83

8.0
7.5
8.5
8.00

9.0
9.0
9.5
9.17

10.0
10.0
10.0
10.00

11.0
11.0
11.0
11.00

0.037

30

60

90

120

150

180

210

240

270

300

Mean
ROR[cm3/sec]

0.0
0.0
0.0
0.00

2.5
2.5
2.5
2.50

3.0
3.0
3.0
3.00

3.5
3.5
3.0
3.33

4.0
4.0
3.5
3.83

4.0
4.5
3.5
4.00

4.5
4.5
4.0
4.33

4.5
5.0
4.0
4.50

4.5
5.0
4.5
4.67

4.5
5.0
4.5
4.67

4.5
5.0
5.0
4.83

0.016

30

60

90

120

150

180

210

240

270

300

Mean
ROR[cm3/sec]

0.0
0.0
0.0
0.0
0.00

3.0
3.0
1.0
3.0
3.00

4.0
3.0
1.0
3.5
3.50

4.5
4.0
1.0
4.0
4.17

4.5
5.0
1.5
5.0
4.83

5.0
6.0
1.5
5.5
5.50

5.0
6.5
1.5
6.0
5.83

5.5
7.0
1.5
6.0
6.17

5.5
7.5
1.5
6.0
6.33

5.5
7.5
1.5
6.5
6.50

6.0
8.0
1.5
6.5
6.83

0.023

30

60

90

120

150

180

210

240

270

300

Mean
3
ROR[cm /sec]

0.0
0.0
0.0
0.00

3.0
0.5
2.5
2.00

4.5
3.5
3.5
3.83

5.5
4.0
4.0
4.50

6.0
5.0
4.5
5.17

7.0
6.0
5.0
6.00

8.0
6.5
5.5
6.67

8.5
7.0
6.0
7.17

9.0
7.5
7.0
7.83

9.5
9.0
7.5
8.67

10.0
10.0
8.0
9.33

0.031

30

60

90

120

150

180

210

240

270

300

Mean
ROR[cm3/sec]

0.0
0.0
0.0
0.0
0.00

2.5
3.0
2.5
5.0
2.67

3.5
4.5
3.5
6.0
3.83

4.0
5.5
4.0
7.0
4.50

4.5
6.0
4.5
9.0
5.00

5.5
6.5
5.0
11.0
5.67

6.5
7.5
6.0
13.0
6.67

7.0
8.0
6.5
14.5
7.17

7.5
8.5
7.5
15.0
7.83

8.5
9.0
8.5
15.0
8.67

9.5
10.0
10.0
15.0
9.83

0.033

Time [sec]

Control

Trial 1
Trial 2
Trial 3
Mean

Vol. of
CO2[cm3]

Time [sec]

pH 1

Trial 1
Trial 2
Trial 3
Mean

Vol. of
CO2[cm3]

Time [sec]

pH 2

Vol. of
CO2[cm3]

Trial 1
Trial 2
Trial 3
Trial 4
Mean

Time [sec]

pH 3

Vol. of
3
CO2[cm ]

Trial 1
Trial 2
Trial 3
Mean

Time [sec]

pH 4

Vol. of
CO2[cm3]

Trial 1
Trial 2
Trial 3
Trial 4
Mean

*Anomalous trials are highlighted

30

60

90

120

150

180

210

240

270

300

Mean
ROR[cm3/sec]

0.0
0.0
0.0
0.00

1.5
2.0
2.0
1.83

2.0
2.5
2.5
2.33

2.5
3.0
3.0
2.83

3.0
3.5
3.5
3.33

3.5
4.0
4.0
3.83

4.5
4.5
4.5
4.50

5.5
5.0
5.0
5.17

6.0
6.0
6.0
6.00

6.5
6.5
6.5
6.50

7.0
7.0
7.0
7.00

0.023

30

60

90

Time [sec]

pH 5

Vol. of
CO2[cm3]

Trial 1
Trial 2
Trial 3
Mean

Time [sec]

pH 6

Vol. of
CO2[cm3]

Trial 1
Trial 2
Trial 3
Mean

Time [sec]

pH 7

Vol. of
CO2[cm3]

Trial 1
Trial 2
Trial 3
Mean

Time [sec]

pH 8

Vol. of
CO2[cm3]

Trial 1
Trial 2
Trial 3
Mean

Time [sec]

pH 9

Vol. of
CO2[cm3]

Trial 1
Trial 2
Trial 3
Mean

Time [sec]

pH 10

Vol. of
CO2[cm3]

Trial 1
Trial 2
Trial 3
Mean

120

150

180

210

240

270

300

0.0 2.0 2.5 3.5 3.5 4.0 4.5 5.0 5.0 5.5 6.0
0.0 2.0 2.5 3.0 3.0 4.0 4.0 4.5 5.0 5.5 6.0
0.0 2.0 2.5 3.5 3.5 4.0 4.0 4.5 4.5 5.0 5.5
0.00 2.00 2.50 3.33 3.33 4.00 4.17 4.67 4.83 5.33 5.83

Mean
ROR[cm3/sec]
0.019

30

60

90

120

150

180

210

240

270

300

Mean
ROR[cm3/sec]

0.0
0.0
0.0
0.00

1.0
0.5
0.5
0.67

1.0
1.0
1.0
1.00

1.5
1.0
1.0
1.17

1.5
1.0
1.0
1.17

2.5
1.5
1.5
1.83

2.5
1.5
2.0
2.00

2.5
2.0
2.0
2.17

3.0
2.5
2.5
2.67

3.0
2.5
2.5
2.67

3.0
3.0
3.0
3.00

0.010

30

60

90

120

150

180

210

240

270

300

Mean
ROR[cm3/sec]

0.0
0.0
0.0
0.00

1.5
1.0
1.0
1.17

2.0
1.0
1.5
1.50

2.0
1.5
1.5
1.67

2.0
1.5
1.5
1.67

2.0
2.0
2.0
2.00

2.0
2.0
2.0
2.00

2.0
2.0
2.0
2.00

2.0
2.0
2.5
2.17

2.0
2.5
2.5
2.33

2.5
2.5
2.5
2.50

0.008

30

60

90

120

150

180

210

240

270

300

Mean
ROR[cm3/sec]

0.0
0.0
0.0
0.00

1.5
1.0
1.0
1.17

1.5
1.0
1.5
1.33

1.5
1.5
1.5
1.50

1.5
1.5
1.5
1.50

1.5
1.5
1.5
1.50

1.5
1.5
1.5
1.50

1.5
1.5
1.5
1.50

1.5
2.0
1.5
1.67

1.5
2.0
2.0
1.83

2.0
2.0
2.0
2.00

0.007

30

60

90

120

150

180

210

240

270

300

Mean
ROR[cm3/sec]

0.0
0.0
0.0
0.00

1.0
0.5
0.5
0.67

1.0
0.5
0.5
0.67

1.0
0.5
0.5
0.67

1.0
1.0
1.0
1.00

1.5
1.0
1.0
1.17

1.5
1.5
1.0
1.33

1.5
1.5
1.0
1.33

1.5
1.5
1.5
1.50

1.5
1.5
1.5
1.50

1.5
1.5
1.5
1.50

0.005

Table 1 Effect of pH on mean rate of reaction:

pH
Control
1
2
3
4
5
6
7
8
9
10

Mean Rate
of Reaction
in cm3/sec
0.037
0.016
0.023
0.031
0.033
0.023
0.019
0.010
0.008
0.007
0.005

The mean rate of reaction was calculated using the equation:

In this case, the mean volume was first calculated and the final mean volume was divided by 300
seconds to find the mean rate of reaction.
In the case of trials where anomalous results were recorded, the entire trial was repeated again
and the anomalous trial was excluded from calculations. The entire trial had to be repeated as if
one result is affected, the rate of reaction for the entire trial changes drastically. Dealing with
anomalies in this manner made the mean results reliable. Anomalous trials may have occurred
due to improper activation of the yeast at an appropriate temperature. If the activation
temperature was too high, the enzymes in yeast could have denatured and if the temperature was
too low, the yeast would have not activated properly.
Word Count: 334

Influence of pH on the function of yeast Interpretation & Evaluation

Statistics:
I will use a Spearman rank correlation statistical test to prove whether my null hypothesis or
alternate hypothesis is being supported.
Sample
pH
Mean ROR
Rank
Difference
Square of diff
Add sq. of
diff.
Rs Value

1
0.016
5
4.98
24.84

2
0.023
8
7.98
63.74

3
4
0.031 0.032
10
9
9.98 8.97
99.54 80.44

5
0.023
7
6.97
48.56

6
0.019
6
5.98
35.72

7
0.010
4
3.98
15.84

8
0.008
3
2.99
8.95

I
9
0.007
2
1.99
3.97

J
10
0.005
1
1.00
0.99

382.60
1.32

Null Hypothesis
There is no correlation present between pH and the rate of respiration of yeast.
Alternate Hypothesis
A correlation is present between pH and the rate of respiration of yeast.
For a significant level of 0.05 probability, the critical value for a two-tailed test is at 0.6485.
The Rs value is the calculated value using the spearman rank formula:
= 1

6 2
(2 1)

where n represents the number of samples.

Since the Rs value, 1.32 is greater than the critical value of 0.6485- we must reject the null
hypothesis and accept the alternate hypothesis that a correlation is present between pH and the
rate of respiration of yeast.
The null hypothesis is rejected for Probability<0.05.

Word Count: 172

Data Analysis:
The result of the Spearman rank correlation test supports the idea that a correlation is present
between pH and the rate of respiration in yeast. As the pH increases or decreases, the rate of
reaction will vary. This is because at a significant level of 0.05 probability, the critical value for a
two-tailed test is at 0.6485. The calculated Rs value [1.32] is greater than the critical by a
difference of 0.6715. The percentage difference between the two values when calculated was at
nearly 104% - this clearly shows that my alternate hypothesis is being supported.
Graph 2 shows a trend in the change in rate of reaction as pH changes. As the pH increases from
1 to 4, there is an increase in the rate of reaction from 0.016cm3/sec to 0.033cm3/sec by a
percentage increase of 51.5%. At pH 4, the largest rate of reaction occurs this can be given as
the optimum pH. As the pH increases from 4 to pH 10, there is a sharp decrease in the rate of
reaction from 0.033cm3/sec to 0.005cm3/sec with a percentage decrease of 84.8%. The
percentage decrease is much larger compared to the percentage increase by a difference of
33.3%. In comparison with the graph in Figure 1, the dumbbell shaped curve coincides with the
relationship shown in my graph thus, they both show similar patterns. The negative correlation
is more dominant compared to the positive and hence, the line of best fit shows an overall
negative correlation between pH and rate of reaction. The presence of a correlation does not
mean that a change in pH causes a change in the rate of reaction; the results only show the
presence of a relationship between the two variables.
Table 2 shows the standard deviation which was calculated using the equationThe standard deviation identifies a range of results that should be obtained if the experiment is
repeated again by another scientist. For example, for pH 1, when repeated, the rate of reaction
should vary within 0.0160.0022cm3/sec. Hence, standard deviation shows a range of possible
errors in the measurement. The greatest standard deviation occurs for pH 4, where the rate of
reaction can vary from 0.0330.0132cm3/sec, so the greatest range of error occurs for pH 3
which is still significantly small.
Standard error of a measurement is an estimate of how often you can expect errors of a given
size. The greater the standard error, the greater the chance of errors and hence the less reliable
the result. The table shows that pH 4 again has the greatest standard error at 0.0076; hence it is
the least reliable from all the results.
The scientific reasoning behind my results can be explained through the working of enzymes.
Yeast contains numerous enzymes, most of which take part in anaerobic respiration for the
process of alcoholic fermentation. [Source 3] Enzymes are proteins and hence, are made up of
long chain amino acids that are folded into a specific shape. The specific and precise folding of
the amino acids results in the side chains of few amino acids forming an active site that can
catalyze or anabolize chemical reactions. Intermolecular forces are present and are responsible
for the stable shape of the protein. One such force is caused by the electrostatic interactions
between positively charged amino acid chains. pH can be a significant factor in determining the
shape of a protein because at different pH values, certain amino acid side chains can become
charged or uncharged this is because pH is determined by the number of H+ ions present. This
is what causes a change in charge which could cause a change in the specific shape of the
protein.

At pH values far from the optimum pH, many important amino acid chains will have the wrong
charge; hence the enzyme will not fold into the specific shape and will lose the ability to bind to
a specific substrate denaturing the enzyme.
[Source 5] Firstly, yeast contains an enzyme called invertase which causes the breakdown of
sucrose to glucose and fructose. Invertase is said to have an optimum pH of 4.5 which is almost
similar to the optimum pH of yeast [pH 4]. At the optimum pH, the maximum amount of sucrose
is broken down, causing the maximum amount of glucose to be produced.
Next, under the lack of oxygen, yeast can respire anaerobically through a process known as
alcoholic fermentation which causes the conversion of glucose to pyruvic acid and then finally to
ethanol and carbon dioxide. In order to produce pyruvic acid from glucose in glycolysis, 10
enzymes are required while in order to produce ethanol and carbon dioxide from pyruvic acid, 2
enzymes are required.
In total, 13 enzymes are required to produce carbon dioxide from sucrose through the anaerobic
respiration of yeast. When pH changes, the specific shape of the active site of these enzymes
changes due to a change in charge, which in turn causes a decrease in the rate at which enzymesubstrate complexes are formed causing a decrease in the rate of respiration of yeast.
Word Count: 845

10

Influence of pH on the function of yeast Communicating

Results:
Table 2 Effect of pH on mean rate of reaction with standard deviation and standard error:

pH
Control
1
2
3
4
5
6
7
8
9
10

Mean Rate
of
Reaction in
cm3/sec
0.037
0.016
0.023
0.031
0.033
0.023
0.019
0.010
0.008
0.007
0.005

Standard
deviation
0.0000
0.0022
0.0028
0.0040
0.0132
0.0000
0.0042
0.0000
0.0022
0.0000
0.0000

Standard
Error
0.0000
0.0013
0.0016
0.0023
0.0076
0.0000
0.0024
0.0000
0.0013
0.0000
0.0000

11

Effect of pH on rate of reaction

0.040

Rate of Reaction in cm3/sec

0.035
0.030
0.025
0.020
0.015
0.010
0.005
0.000
0

10

12

pH

Graph 1 Effect of pH on the rate of reaction of yeast

Figure 2: A typical rate of reaction pH graph

Source: https://y12hb.wordpress.com/2013/03/26/factors-that-effect-enzyme-activity/
This figure was obtained from an educational blog targeted at Year 12 students. It is considered
an unreliable source of data; however the figure is only used for a simple comparison, and not to
provide data for a detailed comparison hence its use can be justified.

12

Evaluation:
Theoretically, I had aimed to keep temperature and the rate at which the conical flask was
swirled at constant, throughout the experiment but practically, doing so proved difficult. The
temperature of the water bath fluctuated frequently within a range of 30oC to 40oC, the rate at
which enzyme-substrate complexes form is faster as temperature increases slightly - which could
have caused a few significant differences in the readings.
Secondly, there could have been differences in the rate at which the conical flask was swirled at.
If the conical flask is swirled faster, the enzyme-substrate catabolic reactions occur faster and
hence the rate of reaction would be faster, causing significant differences between readings.
Even though I had aimed to control variables such as temperature and swirling rate, small
differences in these variables could have caused small errors in my readings these errors are
represented through standard deviation and standard error calculations. [Table 2]
A small sample size was taken as each trial took a significant amount of time and required
thorough preparation. However, even though only a small sample size was used, I conducted 3
trials for each sample and calculated a mean result. Hence, my results are reliable, giving strong
evidence which supports my conclusion that a correlation is present between rate of reaction and
pH.
Modifications could be made to my method which would further increase the validity of my
experiment. Instead of manually adjusting the temperature of the water bath and manually
swirling the conical flask, the use of a digital water bath and digital stirrer would have been
better at controlling temperature and swirling rate. Furthermore, trials could have been conducted
at inter-mediate values of pH such as pH3.6, 4.2, 4.4, etc. in order to find the exact optimum pH
for yeast respiration.
Word Count: 392

13

Sources:
Source 1: Effect of pH on enzyme activity. Brooklyn College. Web page.
Available at: http://academic.brooklyn.cuny.edu/ - Date accessed 19/01/2015
Source 2: How pH affects brewing. Braukaiser Beer. Web page. Last updated in September
2009. Available at: http://www.braukaiser.com Date accessed- 20/01/2015
Source 3: Effects of temperature, pH and sugar concentration on the growth of Saccharomyces
cerevisiae. Published on 25th January 2009.
Journal International Journal of Food Microbiology by F. No Arroyo-Lpez, Sandi Orli,
Amparo Querol, Eladio Barrio
Available at:
http://www.researchgate.net/publication/24043365_Effects_of_temperature_pH_and_sugar_conc
entration_on_the_growth_parameters_of_Saccharomyces_cerevisiae_S._kudriavzevii_and_their
_interspecific_hybrid - Date accessed- 20/01/2015
Source 4: Author- Willie Prejean. Baking Science, The Bakery Network. Web page.
Available at: http://www.thebakerynetwork.com/ - Date accessed 16/01/2015
Source 5: John Wiley & Sons, April 8, 1998. Book.
Yeast Physiology and Biotechnology by Graeme M. Walker.
Available at:
https://books.google.ae/books?id=8rR6Prg3TcC&dq=ph+of+yeast&source=gbs_navlinks_s
Date accessed 16/01/2015
Source 6: CRC Press, December 7, 1990. Book.
Use of Yeast Biomass in Food Production by Anna Halasz, Radomir Lasztity.
Available at:
https://books.google.ae/books?id=xbZRt2pKwqUC&dq=ph+of+yeast&source=gbs_navlinks_s
Date accessed 16/01/2015
I have selected a range of sources varying from websites to books and journals. Source 1 is a
website based on an educational institute known as Brooklyn College. This source is reliable as
the information published on the site is by professors and professionals in their respective fields.
Sources 2 and 4 are the webpages of 2 commercial companies. The baking network and
Braukaiser are on-line companies that specialise in helping small to large industry companies.
These sites are not very reliable but the information referenced was of basic relevance to my
experiment.
Majority of my information was extracted from sources 3, 5 and 6. Source 3 is a collection of
journal papers that were cited by multiple professionals in the field while sources 5 and 6 are
scientific books which would have been reviewed by other scientists in the scientific community
before being published hence these sources are reliable.
Word Count: 150

Total Word Count: 3407

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