Name: ______________________________ Period: ______________ Living Environment Lab

LAB _____. FERMENTATION OF SUCROSE

(aka MAKING ROOT BEER)

All organisms need energy to live. Cellular respiration is the process they use to convert the energy stored in sugars into the quick energy of ATP. If oxygen is available, the mitochondria can perform their “energy generator” job and make a lot of ATP energy. This version of respiration is called aerobic respiration and it produces enough ATP energy to support large active, multicellular animals like you and me.

If oxygen is not available, large organisms cannot produce enough energy to survive. That’s why we die if we cannot breathe. But even though there is no oxygen, some one-celled organisms can still digest sugars and make enough ATP energy to live and grow. This version of respiration is called anaerobic respiration. Anaerobic means “without oxygen”. Anaerobic respiration is used by bacteria and fungi and is also referred to as fermentation. There are two types of fermentation:

•	lactic acid fermentation which is used by bacteria (and how we make yogurt) and also occurs in muscle cells when they are oxygen-deprived like during a sprint race: glucose → ATP + lactic acid 

•	alcoholic fermentation, which is used by yeast (a one-celled fungus) and how we make beer, wine, bread, and many other foods: 
glucose → ATP + alcohol + CO2

In this lab, we are going to explore alcoholic fermentation. Really? Yes, really!

When yeast break down glucose to make ATP energy they also make two waste products: alcohol and CO2. Unfortunately for the yeast the alcohol eventually builds up and kills them. But unlike the yeast, humans see the alcohol as a desirable product — an entertaining beverage. 

We like it so much, we purposely give the yeast sugars to ferment so they make alcohol. The CO2 the yeast also produce collects in the fermenting liquid and makes it fizzy. That’s why we use the terms “carbonation” or “carbonated beverage”. This is the old-fashioned way that soda was made, like root beer, birch beer, and sarsaparilla. And that’s what we are going to recreate in this lab: producing carbonated root beer through the fermentation of sugar

HISTORY OF ROOT BEER:

Root beer was made by our ancestors by soaking Sassafras (a type of tree) root in water, and adding sugar and yeast (for carbonation). In the early 1900s however, scientists discovered that safrole, a chemical found in Sassafras root, was a carcinogen (cancer-causing agent) and human consumption has been banned. Now, a mixture of other herbs and spices makes up “root beer extract” which is what is now used to make homemade root beer.

Materials:
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•	empty 2 liter plastic bottle
•	2 bottles (.5 liter) spring water
•	brewer’s (champagne) yeast
•	sucrose (sugar)
•	root beet extract
•	small cup & large cup
•	funnel
•	graduated cylinder

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PROCEDURE:

Each of you will be given a bottle of spring water. This will be the water that you make your root beer out of and you will pour your root beer back into these bottles to ferment. Your team of two people will be mixing your root beer in one empty 2 liter bottle and then after it is mixed you will pour your soda back into the 0.5 liter spring water bottles. Each student will have their own 0.5 liter bottle of soda at the end of this process.

The procedures listed below are measurements for one lab team of 2 people.

1.	Obtain your warmed bottles of spring water. The bottles should be heated to about body temperature (37°C). 

2.	Measure 0.25g of dry yeast in a small beaker or cup. Add 50mL of the warm water from your spring water bottles to the yeast so it dissolves. Let stand for at least 5 minutes. 

3.	While the yeast is dissolving, measure 4 milliliters of root beer extract. 

4.	Also, measure 110 grams of sucrose (table sugar). 

5.	Get your clean, empty large 2-liter soda bottle. This will be your mixing bottle. First, pour the sucrose into the 2-liter bottle using a funnel. 

6.	Next pour the root beer extract into the same 2-liter bottle. Use some of the warmed spring water to rinse out the root beer extract from the graduated cylinder and add it to the 2-liter bottle. 

7.	Now add the dissolved yeast mixture to the same 2-liter bottle. 

8.	Now add the rest of the 2 bottles of warmed spring water into the same 2-liter bottle to dissolve the sugar and mix everything up gently. You now have enough soda for two people. 

9.	Pour the root beer mixture into your empty spring water bottles. Do not fill the bottle all the way. Only fill to the point that the straight sides of the bottle start to curve in to the neck. Give your teacher any excess root beer mixture to make some ”Tester” bottles. 

10.	Close the cap on your bottle tightly and hold it upside down for a minute to check for leaks. Label the cap with your initials and your class period on it. 

11.	Observe and describe the appearance of the root beer on your lab. 

12.	Give the bottle to your teacher. We will age the root beer for 2–3 weeks at room temperature in a dark place. After that we will refrigerate for 1 week. 

13.	Refrigeration will stop the fermentation process and kill the yeast — and stop us from producing alcohol (Sorry!). Be sure to check bottles every day for tightness, if they get too pressurized, they will burst. 

14.	After chilling, get your bottle, open carefully, pour a sample into a cup, and observe and record the appearance of the root beer. Now have a taste. 

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SUMMARY QUESTIONS

1.	Describe the appearance of the root beer before the fermentation process. 

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2.	Why were the yeast necessary in this experiment? 

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3.	Why was the sucrose necessary in this experiment? 

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4.	Why did we heat the water? 

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5.	What is the yeast trying to do with the sugar? 

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6.	What is the CO2 to the yeast? 

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7.	Why do we have to leave the bottle for a few weeks before we drink it? 

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8.	Why do we want the CO2 to collect in the bottle? __________________________________ 

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9.	What is the alcohol to the yeast? _______________________________________________ 

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10.	What is the alcohol to us? _____________________________________________________ 

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11.	Write the formula for aerobic respiration. 

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12.	Are the yeast using aerobic respiration?__________________________________________ 

13.	What process are the yeast using to make energy? 

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14.	Could we survive using this same process? If not, why not? 

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15.	Write the formula for the version of anaerobic respiration called alcohol fermentation. 

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