3.

Anaerobic Respiration in Yeast

Title: Anaerobic respiration in yeast

Aim:  To use a differential respirometer to determine the rate of respiration in

yeast

Apparatus:

·         Respirometer

·         1 1000 mL beaker

·         Laboratory thermometer

·         Paper towels

·         Timer

·         2.5 mL baker’s yeast suspension

·         8 mL of 5% glucose solution

·         2 measuring cylinders (for glucose and for yeast)

·         Paraffin

·         Dropper

·         Water bath at 37-40°C

Method:

1.              Fill a 1000 mL beaker to 900 mL with tap water and allow it to reach
room temperature. Monitor with a laboratory thermometer.

2.              Add 2.5 mL of yeast suspension and 2.5 mL of 5% glucose solution to the
experimental test tube in bracket ‘E’.

3.              Using a dropper, add enough paraffin to cover the surface of the
yeast/glucose solution. Do not allow the paraffin to touch the walls of the test
tube.

4.              Incubate the yeast/glucose solution in the water bath for 10 minutes at
37-40°C to ‘jump start’ the fermentation reaction.
5.              Fill the control test tube in bracket ‘C’ with 5 mL of 5% glucose solution,
then add the same amount of paraffin in the experimental test tube.

6.              Before inserting the stoppers into the test tubes, pull the plunger of the
control-chamber syringe (on left) all the way to the top. Push the plunger of the
experimental-chamber syringe (on right) all the way down.

7.              Insert the stoppers into the mouth of the test tubes.

8.      By pushing the control plunger down and/or pulling the experimental
plunger up, adjust the initial levels of blue-fluid in the control and experimental
pipette so that the control pipette (on left) reads 0.700 mL and the experimental
(on right) reads 0.100 mL.

9.      Place the entire assembly over the edge of the room temperature 1000 mL
beaker water bath that was filled in step 1.

10.  Allow the submerged test tubes to equilibrate in the water bath for a few
minutes and then record the temperature. Make a final adjustment of the
experimental tube to read 0.100 mL and record this as the initial volume at time 0.
As the fermentation of the sugar progresses, the pressure from the carbon dioxide
gas will force the blue liquid to move down the tube.

11.  Record the volume changes at 0.5 minutes (30 seconds) for a minimum of 10-
15 mins. Calculate the carbon dioxide produced per interval by taking the
difference between the initial reading at 0.100 mL and the final reading.

12.  Plot the volume of carbon dioxide (mL) versus time (minutes). The graph
should be linear. The slope (gradient) can be calculated by ∆Y/∆X = (Y2-Y1)/(X2-
X1).

Please see the video below which provides further guidance on the set-up of the
differential respirometer:

https://youtu.be/EhpL1e92hN8

Results:

Change in Volume (mL)

Time (min.)
Group 1 Group 2 Group 3 Group 4
0.00 0.00 0.00 0.00 0.00
0.50 0.04 0.08 0.05 0.05
1.00 0.08 0.16 0.10 0.13
1.50 0.12 0.23 0.14 0.19
 2.00 0.16 0.29 0.18 0.24
2.50 0.20 0.35 0.20 0.29
3.00 0.24 0.42 0.27 0.39
3.50 0.28 0.48 0.32 0.39
4.00 0.32 0.54 0.36 0.44
4.50 0.36 0.60 0.40 0.49
5.00 0.40 0.60 0.44 0.54
5.50 0.44 0.60 0.48 0.60
6.00 0.48 0.60 0.52 0.64
6.50 0.51 0.60 0.56 0.66
7.00 0.56 0.60 0.60 0.66
 

Discussion guidelines: (be sure to address the following in the course of your
discussion):

1. Please note that accurate background information

on anaerobic respiration and on the simple respirometer should be
provided.

2.     From your graph (this should be a single graph with all the plots), describe the
relationship between the amount of carbon dioxide produced versus time for the
glucose fermentation. The trend observed for the groups should be noted and
explained.

3.     What is the rate of carbon dioxide production per minute for the fermentation
of glucose based on your group's results? (The rate is given by the slope of the
graph). What is the average rate of carbon dioxide production per minute for the
fermentation of glucose?

4.     If 2.5 mL of a 5% glucose solution were used in the experimental test tube,
how many grams of glucose would there be in the tube? (Hint: For a 5% solution
there are 5 g of glucose/100 mL of solution.)

5.     Calculate the average fermentation rate per gram of glucose at room

temperature. The fermentation rate can be calculated using the following formula.

Rate per gram = slope/total grams of glucose = _____ mL CO 2/min./g.

6.     What is the purpose of the paraffin used to cover the yeast and glucose
solution in the experimental tube?

7.     What are some factors that may affect the fermentation rate of glucose?
8.     Write the balanced chemical equation for the fermentation of glucose

Please note that this lab is being assessed for MM and AI.