aim of yeast fermentation experiment

The fermentation of sugars using yeast: A discovery experiment

Charles Pepin (student) and Charles Marzzacco (retired), Melbourne, FL

Introduction

Enzyme catalysis 1  is an important topic which is often neglected in introductory chemistry courses. In this paper, we present a simple experiment involving the yeast-catalyzed fermentation of sugars. The experiment is easy to carry out, does not require expensive equipment and is suitable for introductory chemistry courses.

The sugars used in this study are sucrose and lactose (disaccharides), and glucose, fructose and galactose (monosaccharides). Lactose, glucose and fructose were obtained from a health food store and the galactose from Carolina Science Supply Company. The sucrose was obtained at the grocery store as white sugar. The question that we wanted to answer was “Do all sugars undergo yeast fermentation at the same rate?”

Sugar fermentation results in the production of ethanol and carbon dioxide. In the case of sucrose, the fermentation reaction is:

\[C_{12}H_{22}O_{11}(aq)+H_2 O\overset{Yeast\:Enzymes}{\longrightarrow}4C_{2}H_{5}OH(aq) + 4CO_{2}(g)\]

Lactose is also C 12 H 22 O 11  but the atoms are arranged differently. Before the disaccharides sucrose and lactose can undergo fermentation, they have to be broken down into monosaccharides by the hydrolysis reaction shown below:

\[C_{12}H_{22}O_{11} + H_{2}O \longrightarrow 2C_{6}H_{12}O_{6}\]

The hydrolysis of sucrose results in the formation of glucose and fructose, while lactose produces glucose and galactose.

sucrose + water \(\longrightarrow\) glucose + fructose

lactose + water \(\longrightarrow\) glucose + galactose

The enzymes sucrase and lactase are capable of catalyzing the hydrolysis of sucrose and lactose, respectively.

The monosaccharides glucose, fructose and galactose all have the molecular formula C 6 H 12 O 6  and ferment as follows:

\[C_{6}H_{12}O_{6}(aq)\overset{Yeast Enzymes}{\longrightarrow}2C_{2}H_{5}OH(aq) + 2CO_{2}(g)\]

In our experiments 20.0 g of the sugar was dissolved in 100 mL of tap water. Next 7.0 g of Red Star ®  Quick-Rise Yeast was added to the solution and the mixture was microwaved for 15 seconds at full power in order to fully activate the yeast. (The microwave power is 1.65 kW.) This resulted in a temperature of about 110  o F (43  o C) which is in the recommended temperature range for activation. The cap was loosened to allow the carbon dioxide to escape. The mass of the reaction mixture was measured as a function of time. The reaction mixture was kept at ambient temperature, and no attempt at temperature control was used. Each package of Red Star Quick-Rise Yeast has a mass of 7.0 g so this amount was selected for convenience. Other brands of baker’s yeast could have been used.

This method of studying chemical reactions has been reported by Lugemwa and Duffy et al. 2,3  We used a balance good to 0.1 g to do the measurements. Although fermentation is an anaerobic process, it is not necessary to exclude oxygen to do these experiments. Lactose and galactose dissolve slowly. Mild heat using a microwave greatly speeds up the process. When using these sugars, allow the sugar solutions to cool to room temperature before adding the yeast and microwaving for an additional 15 seconds.

Fermentation rate of sucrose, lactose alone, and lactose with lactase

Fig. 1 shows plots of mass loss vs time for sucrose, lactose alone and lactose with a dietary supplement lactase tablet added 1.5 hours before starting the experiment. All samples had 20.0 g of the respective sugar and 7.0 g of Red Star Quick-Rise Yeast. Initially the mass loss was recorded every 30 minutes. We continued taking readings until the mass leveled off which was about 600 minutes. If one wanted to speed up the reaction, a larger amount of yeast could be used. The results show that while sucrose readily undergoes mass loss and thus fermentation, lactose does not. Clearly the enzymes in the yeast are unable to cause the lactose to ferment. However, when lactase is present significant fermentation occurs. Lactase causes lactose to split into glucose and galactose. A comparison of the sucrose fermentation curve with the lactose containing lactase curve shows that initially they both ferment at the same rate.

Fig. 1. Comparison of the mass of CO 2 released vs time for the fermentation of sucrose, lactose alone, and lactose with a lactase tablet. Each 20.0 g sample was dissolved in 100 mL of tap water and then 7.0 g of Red Star Quick-Rise Yeast was added.

However, when the reactions go to completion, the lactose, lactase and yeast mixture gives off only about half as much CO 2  as the sucrose and yeast mixture. This suggests that one of the two sugars that result when lactose undergoes hydrolysis does not undergo yeast fermentation. In order to verify this, we compared the rates of fermentation of glucose and galactose using yeast and found that in the presence of yeast glucose readily undergoes fermentation while no fermentation occurs in galactose.

Fig. 2. Comparison of the mass of CO 2 released vs time for the fermentation of sucrose, glucose and fructose. Each 20 g sugar sample was dissolved in 100 mL of water and then 7.0 g of yeast was added.

Fermentation rate of sucrose, glucose and fructose

Next we decided to compare the rate of fermentation of sucrose with that glucose and fructose, the two compounds that make up sucrose. We hypothesized that the disaccharide would ferment more slowly because it would first have to undergo hydrolysis. In fact, though, Fig. 2 shows that the three sugars give off CO 2  at about the same rate. Our hypothesis was wrong. Although there is some divergence of the three curves at longer times, the sucrose curve is always as high as or higher than the glucose and fructose curves. The observation that the total amount of CO 2  released at the end is not the same for the three sugars may be due to the purity of the fructose and glucose samples not being as high as that of the sucrose.

Fermentation rate and sugar concentration

Next, we decided to investigate how the rate of fermentation depends on the concentration of the sugar. Fig. 3 shows the yeast fermentation curves for 10.0 g and 20.0 g of glucose. It can be seen that the initial rate of CO 2  mass loss is the same for the 10.0 and 20.0 g samples. Of course the total amount of CO 2  given off by the 20.0 g sample is twice as much as that for the 10.0 g sample as is expected. Later, we repeated this experiment using sucrose in place of glucose and obtained the same result.

Fig. 3. Comparison of the mass of CO 2  released vs time for the fermentation of 20.0 g of glucose and 10.0 g of glucose. Each sugar sample was dissolved in 100 mL of water and then 7.0 g of yeast was added.

Fermentation rate and yeast concentration

After seeing that the rate of yeast fermentation does not depend on the concentration of sugar under the conditions of our experiments, we decided to see if it depends on the concentration of the yeast. We took two 20.0 g samples of glucose and added 7.0 g of yeast to one and 3.5 g to the other. The results are shown in Fig. 4. It can clearly be seen that the rate of CO 2  release does depend on the concentration of the yeast. The slope of the sample with 7.0 g of yeast is about twice as large as that with 3.5 g of yeast. We repeated the experiment with sucrose and fructose in place of glucose and obtained similar results.

Fig. 4. Comparison of the mass of CO 2 released vs time for the fermentation of two 20.0 g samples of glucose dissolved in 100 mL of water. One had 7.0 g of yeast and the other had 3.5 g of yeast.

In hindsight, the observation that the rate of fermentation is dependent on the concentration of yeast but independent of the concentration of sugar is not surprising. Enzyme saturation can be explained to students in very simple terms. A molecule such as glucose is rather small compared to a typical enzyme. Enzymes are proteins with large molar masses that are typically greater than 100,000 g/mol. 1  Clearly, there are many more glucose molecules in the reaction mixture than enzyme molecules. The large molecular ratio of sugar to enzyme clearly means that every enzyme site is occupied by a sugar molecule. Thus, doubling or halving the sugar concentration cannot make a significant difference in the initial rate of the reaction. On the other hand, doubling the concentration of the enzyme should double the rate of reaction since you are doubling the number of enzyme sites.

The experiments described here are easy to perform and require only a balance good to 0.1 g and a timer. The results of these experiments can be discussed at various levels of sophistication and are consistent with enzyme kinetics as described by the Michaelis-Menten model. 1  The experiments can be extended to look at the effect of temperature on the rate of reaction. For enzyme reactions such as this, the reaction does not take place if the temperature is too high because the enzymes get denatured. The effect of pH and salt concentration can also be investigated.

• Jeremy M. Berg, John L. Tymoczko and Lubert Stryer,  Biochemistry , 6th edition, W.H. Freeman and Company, 2007, pages 205-237.
• Fugentius Lugemwa, Decomposition of Hydrogen Peroxide,  Chemical Educator , April 2013, pages 85-87.
• Daniel Q. Duffy, Stephanie A. Shaw, William D. Bare, Kenneth A. Goldsby, More Chemistry in a Soda Bottle, A Conservation of Mass Activity,  Journal of Chemical Education , August 1995, pages 734-736.
• Jessica L Epstein, Matthew Vieira, Binod Aryal, Nicolas Vera and Melissa Solis, Developing Biofuel in the Teaching Laboratory: Ethanol from Various Sources,  Journal of Chemical Education , April 2010, pages 708–710.

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Yeast Fermentation Experiment

Fermentation is a fascinating process that kids can easily explore through a simple experiment using yeast and sugar. This hands-on activity teaches students about fermentation and introduces them to the scientific method, data collection, and analysis.

Investigate how different types of sugar (white, brown, and honey) affect the rate of yeast fermentation by measuring the amount of carbon dioxide (CO₂) produced.

Example Hypothesis: If yeast is added to different types of sugar, then the type of sugar will affect the amount of carbon dioxide produced, with white sugar producing more CO₂ than the others.

💡 Learn more about using the scientific method [here] and choosing variables .

Watch the Video:

• Active dry yeast
• White sugar
• Brown sugar
• Measuring spoons and measuring cups
• Small bottles or test tubes
• Rubber bands
• Ruler or measuring tape
• Notebook and pen for recording data ( grab free journal sheets here )
• Printable Experiment Page (see below)

Instructions:

STEP 1. Prepare a yeast solution by dissolving a packet of active dry yeast in warm water according to the package instructions.

STEP 2. Label 3 bottles and add 1 tablespoon of white sugar to the “White Sugar” bottle. Add 1 tablespoon of brown sugar to the “Brown Sugar” bottle. Measure 1 tablespoon of honey and add it to the “Honey” bottle.

STEP 3. Measure and pour an equal amount of the yeast solution into each bottle, ensuring the yeast is well mixed with the sugar.

STEP 4. Quickly stretch a balloon over the mouth of each bottle. Secure the balloons with rubber bands if needed. Ensure the balloons are sealed tightly to prevent CO₂ from escaping.

STEP 5. Place the bottles in a warm, consistent environment to promote fermentation.

STEP 6. Observe and record the size of the balloons at regular intervals (e.g., every 15 minutes) for 1-2 hours. Use a ruler or measuring tape to measure the circumference of each balloon.

TIP: Note the time it takes for the balloons to start inflating and the differences in balloon size over time for each type of sugar.

STEP 7: Analyze the data by comparing the amount of CO₂ produced (balloon size) for each type of sugar. Create a graph showing the balloon size over time for each sugar type.

STEP 8. Determine which sugar type resulted in the most and least CO₂ production. Discuss possible reasons for the differences, considering what each sugar is made of. Think about whether the results support or disprove the hypothesis. Can you come up with further experiments or variations to explore other factors affecting yeast fermentation?

Free Printable Yeast and Sugar Experiment Project

Grab the free fermentation experiment worksheet here. Join our STEM club for a printable version of the video!

The Science Behind Yeast Fermentation

For Our Younger Scientists: Yeast is a type of fungus that feeds on sugars. When you mix yeast with sugar and water, it starts to eat the sugar and convert it into alcohol and carbon dioxide gas. The gas gets trapped in the balloon, causing it to inflate. This shows that fermentation is happening!

Yeast fermentation is a biological process where yeast converts sugars into alcohol and carbon dioxide (CO₂) in the absence of oxygen. This process is used in baking, brewing, wine making and biofuel production. How much fermentation occurs can vary depending on the type of sugar used.

Yeast contains enzymes that break down sugar molecules through a series of chemical reactions . Here’s how it works:

Enzymes are molecules, usually proteins, that act as catalysts to speed up chemical reactions within living organisms.

First the yeast is mixed with warm water, and it becomes activated. The warm environment “wakes up” the yeast cells, preparing them to consume sugars.

Yeast cells produce enzymes that break down sugar molecules (sucrose, glucose, and fructose) into simpler molecules. This process is called glycolysis. During glycolysis, sugar molecules are converted into pyruvate, releasing a small amount of energy.

In the absence of oxygen (anaerobic conditions), yeast cells convert pyruvate into ethanol (alcohol) and carbon dioxide gas (CO₂). The carbon dioxide produced during fermentation is what inflates the balloons in the experiment.

Different Sugars & Fermentation

Different sugars can affect the rate of fermentation. This is how:

• White Sugar (Sucrose): Composed of glucose and fructose and is easily broken down by yeast, leading to efficient CO₂ production.
• Brown Sugar: Contains sucrose along with molasses, which includes minerals and additional nutrients. May result in a slightly different fermentation rate due to its composition.
• Honey: Contains a mixture of glucose, fructose, and other components. The additional components can influence the fermentation process, potentially leading to different CO₂ production rates compared to pure sucrose.

The amount of CO₂ produced depends on how easily the yeast can break down the sugar molecules and convert them into ethanol and CO₂. Sugars that are more readily broken down by yeast will typically produce more CO₂ faster.

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Science project, growing yeast: sugar fermentation.

Yeast is most commonly used in the kitchen to make dough rise. Have you ever watched pizza crust or a loaf of bread swell in the oven? Yeast makes the dough expand. But what is yeast exactly and how does it work? Yeast strains are actually made up of living eukaryotic microbes, meaning that they contain cells with nuclei. Being classified as fungi (the same kingdom as mushrooms), yeast is more closely related to you than plants! In this experiment we will be watching yeast come to life as it breaks down sugar, also known as sucrose , through a process called fermentation . Let’s explore how this happens and why!

What is sugar’s effect on yeast?

• 3 Clear glass cups
• 2 Teaspoons sugar
• Water (warm and cold)
• 3 Small dishes
• Permanent marker

• Fill all three dishes with about 2 inches of cold water
• Place your clear glasses in each dish and label them 1, 2, and 3.
• In glass 1, mix one teaspoon of yeast, ¼ cup of warm water, and 2 teaspoons of sugar.
• In glass 2, mix one teaspoon of yeast with ¼ cup of warm water.
• In glass 3, place one teaspoon of yeast in the glass.
• Observe each cups reaction. Why do you think the reactions in each glass differed from one another? Try using more of your senses to evaluate your three glasses; sight, touch, hearing and smell especially!

The warm water and sugar in glass 1 caused foaming due to fermentation. 

Fermentation is a chemical process of breaking down a particular substance by bacteria, microorganisms, or in this case, yeast. The yeast in glass 1 was activated by adding warm water and sugar. The foaming results from the yeast eating the sucrose. Did glass 1 smell different? Typically, the sugar fermentation process gives off heat and/or gas as a waste product. In this experiment glass 1 gave off carbon dioxide as its waste.

Yeast microbes react different in varying environments. Had you tried to mix yeast with sugar and cold water, you would not have had the same results. The environment matters, and if the water were too hot, it would kill the yeast microorganisms. The yeast alone does not react until sugar and warm water are added and mixed to create the fermentation process. To further investigate how carbon dioxide works in this process, you can mix yeast, warm water and sugar in a bottle while attaching a balloon to the open mouth. The balloon will expand as the gas from the yeast fermentation rises.

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Grow yeast experiment

Follow FizzicsEd 150 Science Experiments:

You Will Need:

• 4 packets of dry yeast
• 4 water bottles, chilled in the fridge (we use Thank You Water, a social enterprise that works to get clean water & sanitation to people in need)
• 1 large jug.
• 4 measuring cups.
• 4 thermometers (one will do if you don’t have a class set).
• Access to boiling water plus adult supervision.
• 1 stopwatch.
• A pen to mark the water temperature on each water bottle during the experiment.
• A shelf to leave the science experiment to run.
• A notebook for your observations.

• Instruction

Pour out the 4 chilled water bottles into the large jug and discard the rest of the water (maybe water your  school garden !)

Carefully measure out the water into the four measuring cups as per the measurements below;

Cup 1 – 200mL of chilled water

Cup 2 – 150mL of chilled water

Cup 3 – 100mL of chilled water

Cup 4 – 50mL of chilled water

Use the thermometers to take a measurement of the water temperature in each cup (write this in your notebook).

With an adult, boil a jug of water and then top up cups 2, 3 and 4 so that they too have 200mL of water as per cup 1. You will be testing the effect of temperature on the growth of yeast by measuring how much gas is released by the yeast under 4 different temperature conditions ( variable testing ).

Using a funnel, carefully pour each cup of water into the four separate water bottles. Use the pen to mark the starting temperature of each water bottle.

Add a spoonful of sugar per water bottle and then swirl the bottle to dissolve the sugar.

Add a yeast packet into each bottle and quickly stretch a balloon of the opening of each bottle.

4 yeast growth experiments started, showing a distinct change already!

Start the stopwatch and take notes of when each balloon rises!

OPTIONAL: you could also keep each bottle in the yeast experiment at the same temperature and vary the amount of sugar added instead.

Go further – buy 4 x student activity sheets as extension worksheets.

This student science booklet has been created by experienced science educators from the Fizzics Education team.

Use these student worksheets as blackline masters for your science class!

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What is going on?

Your experiment was testing the effect of water temperature on the growth of yeast. Yeast are egg-shaped microscopic cells of fungi that are dormant whilst kept in dry and cool conditions. However, yeast will rapidly divide once exposed to water and sugar in ideal temperatures. In the right temperature, yeast cells will change the sugar into glucose by using the water plus as an enzyme catalyst (invertase). Once the yeast has converted the sugar to glucose fermentation can then occur to produce carbon dioxide and ethanol as per the equation below;

Glucose ⟶ Ethanol + Carbon dioxide

which can be written as…

C 6 H 12 O 6(aq)   ⟶ 2C 2 H 5 OH (aq)  + 2CO 2(g)

In your experiment, you were trapping the carbon dioxide released during the fermentation process. The more active the yeast, the more carbon dioxide the yeast produced! In your experiment, the different water temperatures will have produced different results as bottles may have been too hot for the yeast to survive whereas the other bottles may have been too cold.   By introducing a variable to test in your experiment, you’re doing real science!  The following list of temperatures is worth keeping in mind when assessing your results:

• 55° C – 60° C Yeast cells die (also known as the thermal death point).
• 41° C – 46° C Ideal temperature of water for dry yeast being reconstituted with water and sugar.
• 4° C The temperature of a fridge – yeast will be too cold to work properly.

Yeast is used to make bread rise and to ferment beer. There are many different species of yeast, but the one most commonly used in cooking and baking is called  Saccharomyces cerevisiae , which is also known as brewer’s yeast.

Yeast can break down many types of simple carbohydrates (monosaccharides) however they cannot break down complex carbohydrates such as starch. This means that extra enzymes are needed to break down starch into sugars that the yeast can use, for example during beer production we use enzymes from germinating barley to do this.

Variables to test

More on variables here

• Try different concentrations of vinegar as the growth medium. Can the yeast handle some acidity?
• Vary the amount of sugar used.
• Does the volume of water make a difference?

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4 thoughts on “ Grow yeast experiment ”

What is the amount of water you would like me to put?

Hi! Here’s the detail’s that you need;

> Cup 1 – 200mL of chilled water > Cup 2 – 150mL of chilled water > Cup 3 – 100mL of chilled water > Cup 4 – 50mL of chilled water

With an adult, boil a jug of water and then top up cups 2, 3 and 4 so that they too have 200mL of water as per cup 1. You will be testing the effect of temperature on the growth of yeast by measuring how much gas is released by the yeast under 4 different temperature conditions.

Would this experiment still work if instead i tested how different types of sugars affect the amount of fermentation by yeast. Would i still get different sized balloons in my result.

We’d love it if you try this and let us know! With any experiment you just have to change one thing and then measure the result. So, changing the types of sugars is a completely valid investigation. Good luck!

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3.1.3 Yeast experiment explained

You’ve seen the results of the yeast experiment, but what do these results mean?

Yeasts are microscopic, single-celled organisms, and are a type of fungus that is found all around us, in water, soil, on plants, on animals and in the air. Like all organisms, when yeasts are put in the right type of environment they will thrive; growing and reproducing.

Your experiments were designed to help you identify which environment promotes the most yeast growth. The first three glasses in your experiment contained different temperature environments (cold water, hot water and body temperature water). At very low temperatures the yeast simply does not grow but it is still alive – if the environment were to warm up a bit, it would gradually begin to grow. At very high temperatures the cells within the yeast become damaged beyond repair and even if the temperature of that environment cooled, the yeast would still be unable to grow. At optimum temperatures the yeast thrives.

Your third and fourth glasses both contained environments at optimum temperature (body temperature) for yeast growth, the difference being, the fourth glass was sealed. The variable between these two experiments was the amount of available oxygen. You may have been surprised by your results here, thinking that a living organism in an environment without oxygen cannot survive? However, you should have found that yeast grew pretty well in both experiments.

To understand why yeast was able to thrive in both conditions we need to understand the chemical process occurring in each glass during the experiment. In the three open glasses, oxygen is readily available, and from the moment you added the yeast to the sugar solution it began to chemically convert the sugar in the water and the oxygen in the air into energy, water, and carbon dioxide in a process called aerobic respiration.

Yeast is a slightly unusual organism – it is a ‘facultative anaerobe’. This means that in oxygen-free environments they can still survive. The yeast simply switches from aerobic respiration (requiring oxygen) to anaerobic respiration (not requiring oxygen) and converts its food without oxygen in a process known as fermentation. Due to the absence of oxygen, the waste products of this chemical reaction are different and this fermentation process results in carbon dioxide and ethanol.

Depending on how long you monitored your experiment for and how much space your yeast had to grow you may have noticed that, with time, the experiment sealed with cling film slowed down. This is for two reasons; firstly because less energy is produced by anaerobic respiration than by aerobic respiration and, secondly, because the ethanol produced is actually toxic to the yeast. As the ethanol concentration in the environment increases, the yeast cells begin to get damaged, slowing their growth.

The ethanol produced is a type of alcohol, so it is this process that allows us to use it to make beer and wine. When used in bread making, the yeast begins by respiring aerobically, the carbon dioxide from which makes the bread rise. Eventually the available oxygen is used up, and the yeast switches to anaerobic respiration producing alcohol and carbon dioxide instead. Do not worry though; this alcohol evaporates during the baking process, so you won’t get drunk at lunchtime from eating your sandwiches.

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• Topic: Microbes

The Role of Yeasts in Fermentation Processes

In recent years, vessels have been discovered that contain the remains of wine with an age close to 7000 years. It is unclear whether, in ancient times, humans accidentally stumbled across fermented beverages like wine or beer, or was it a product intended as such. What is a fact is that since then, alcoholic beverages have been part of the diet and culture of many of the civilizations that have preceded us. The typical examples of beer and wine are an example of many other drinks resulting from the action of yeasts. In addition to these two beverages, various companies have developed other types of fermented foods and non-alcoholic beverages prepared in a traditional or commercial manner. The climatic conditions, the availability of raw material and the preferences of each region have conditioned and favored the maintenance of some of these products. In addition to the aforementioned traditional alcoholic beverages produced from fruits, berries, or grains, humans use yeast in the production of chemical precursors, global food processing such as coffee and chocolate, or even wastewater processing. Yeast fermentation is not only useful in food manufacturing. Its uses extend to other products of high interest such as the generation of fuel from vegetable sources.

1. Introduction

Fermentation is a well-known natural process used by humanity for thousands of years with the fundamental purpose of making alcoholic beverages, as well as bread and by-products. Upon a strictly biochemical point of view, fermentation is a process of central metabolism in which an organism converts a carbohydrate, such as starch or sugar, into an alcohol or an acid. For example, yeast performs fermentation to obtain energy by converting sugar into alcohol. Fermentation processes were spontaneously carried out before the biochemical process was fully understood. In the 1850s and 1860s, the French chemist and microbiologist Louis Pasteur became the first scientist to study fermentation, when he demonstrated that this process was performed by living cells. Fermentation processes to produce wines, beers and ciders are traditionally carried out with Saccharomyces cerevisiae strains, the most common and commercially available yeast. They are well known for their fermentative behavior and technological characteristics which allow obtaining products of uniform and standard quality. Many other important industrial products are the result of fermentation, such as yogurt, cheese, bread, coffee. Yeasts also play a key role in wastewater treatment or biofuel production. Upon a biochemical point of view, fermentation is carried out by yeasts (and some bacteria) when pyruvate generated from glucose metabolism is broken into ethanol and carbon dioxide ( Figure 1 ).

Central metabolism of fermentation in yeasts.

The schematic chemical equation for the production of ethanol from glucose is as follows:

Under absence or oxygen-limited conditions, ethanol is produced from acetaldehyde, and two moles of ATP are generated. This is not a fully satisfactory reaction for cells, as they have to consume high amounts of glucose to deliver enough ATP to the system. As a consequence, ethanol is accumulated and when this occurs the fermentative activity is stopped [ 1 ].

1.1. Yeasts

Yeasts are eukaryotic microorganisms that live in a wide variety of ecological niches, mainly in water, soil, air and on plant and fruit surfaces. Perhaps the most interesting habitat at this point is the latter, since they directly intervene in the decomposition of ripe fruit and participate in the fermentation process. In this natural environment, yeasts can carry out their metabolism and fermentation activity satisfactorily as they have the necessary nutrients and substrates [ 2 ]. On a nutritional level, yeasts are not particularly demanding compared to other microorganisms such as lactic acid bacteria. However, their growth is supported by the existence of basic compounds such as fermentable sugars, amino acids, vitamins, minerals and also oxygen. Upon a morphological point of view, yeasts present a high morphological divergence, with round, ellipsoidal and oval shapes being the most common. In fact, in the identification processes, microscopic evaluation is the first resource followed by other more discriminatory tests such as microbiological and biochemical ones. In a next stage, the classical classification includes other more laborious tests such as those of sugar fermentation and amino acid assimilation [ 2 ]. The production and tolerance to ethanol, organic acids and SO 2 are also important tools to differentiate among species. The reproduction of yeasts is mainly by budding, which results in a new and genetically identical cell. Budding is the most common type of asexual reproduction, although cell fission is a characteristic of yeasts belonging to the genus Schizosaccharomyces . Growing conditions that lead to nutrient starvation, such as lack of amino acids, induce sporulation, which is a mechanism used by yeasts to survive in adverse conditions. As a result of sporulation, yeast cells suffer from genetic variability. In industrial fermentation processes, the asexual reproduction of yeasts is advisable to ensure the preservation of the genotype and to maintain stable fermentation behaviour that does not derive from it for as long as possible. At the metabolic level, yeasts are characterised by their capacity to ferment a high spectrum of sugars, among which glucose, fructose, sucrose, maltose and maltotriose predominate, found both in ripe fruit and in processed cereals. In addition, yeasts tolerate acidic environments with pH values around 3.5 or even less. According to technological convenience, yeasts are divided into two large groups namely Saccharomyces and non- Saccharomyces . Morphologically, Saccharomyces yeasts can be round or ellipsoidal in shape depending on the growth phase and cultivation conditions. S. cerevisiae is the most studied species and the most utilized in the fermentation of wines and beers due to its satisfactory fermentative capacity, rapid growth and easy adaptation. They tolerate concentrations of SO 2 that normally most non- Saccharomyces yeasts do not survive. However, despite these advantages, it is possible to find in the nature representatives of S. cerevisiae that do not necessarily have these characteristics.

1.2. Non- Saccharomyces Yeasts

Non- Saccharomyces yeasts are a group of microorganisms used in numerous fermentation processes, since their high metabolic differences allow the synthesis of different final products. Generally, many of these yeasts capable of modifying the sensory quality of wines are considered as contaminants, so eliminating them or keeping them at low levels was a basic objective in the past [ 3 ]. In order to eliminate their activity in wine fermentation, it is usual to disinfect the tanks and fermentation containers using sulfite. This perception has been modified year after year, gaining relevance the action of these yeasts in the spontaneous fermentation, since they contribute positively in the final sensory quality of the wine. These yeasts are the majority in the initial phase of spontaneous fermentation to the point where the concentration of ethanol reaches 4 and 5% v / v . At that point, between alcohol and the exhaustion of dissolved oxygen, their growth is inhibited [ 4 ]. When the process is completed, Saccharomyces yeasts, the most resistant to ethanol, predominate and complete the fermentation. It has been reported that some non- Saccharomyces yeasts are able to survive toward the end of the spontaneous fermentation and exert their metabolic activity, thus contributing positively to the sensory quality of wines. Based on this evidence, in recent years, many researchers have focused their studies in understanding the nature and fermentative activity of the non- Saccharomyces yeasts [ 5 ]. The findings demonstrated the enormous potential of these yeasts for use in the fermentation of traditional and nontraditional beverages. Despite the fact that most non- Saccharomyces yeasts show some technological disadvantages compared to S. cerevisiae such as lower fermentative power and production of ethanol, non- Saccharomyces yeasts possess characteristics that in S. cerevisiae are absent, for instance, production of high levels of aromatic compounds such as esters, higher alcohols and fatty acids [ 6 ]. In addition, it has been reported that the fermentative activity of these yeasts is manifested in the presence of small amounts of oxygen which leads to an increase in cell biomass and the decrease in ethanol yield, a strategy that can be used to reduce the ethanol content of wines produced in coculture with S. cerevisiae [ 7 ]. With the aim of exploiting the positive characteristics of non- Saccharomyces yeasts and reducing their negative impact, fermentations with mixed and sequential cultures with S. cerevisiae can be performed to produce fermented beverages with different sensory profiles [ 8 ]. The most important fact is related to the potential for producing a broad variety of compounds of sensory importance necessary to improve the organoleptic quality of wines and beers. The findings reported so far in literature have led to rethink the role of these yeasts in fermentative processes and to evaluate their use in the development of new products. Among the most studied non- Saccharomyces yeasts that reached special importance for researchers include Candida , Kloeckera , Hanseniaspora , Brettanomyces , Pichia , Lanchacea and Kluyveromyces , among others.

2. Yeast Fermentation Processes

2.1. alcoholic fermentations.

The production of alcoholic beverages from fermentable carbon sources by yeast is the oldest and most economically important of all biotechnologies. Yeast plays a vital role in the production of all alcoholic beverages. Yeast plays a vital role in the production of all alcoholic beverages and the selection of suitable yeast strains is essential not only to maximise alcohol yield, but also to maintain beverage sensory quality [ 2 ].

2.1.1. Wine Fermentation

In wine fermentation, strains with specific characteristics are needed, for instance, highly producers of ethanol to reach values of 11–13% v / v , typically found in this beverage. On the other hand, beers and ciders contain less amounts of ethanol with a balanced and distinctive sensory profile characteristic of each one. In recent years, new consuming trends and requirements for new and innovative products have emerged. This situation led to rethink about the existing fermented beverages and to meet the demands of consumers. Yeasts are largely responsible for the complexity and sensory quality of fermented beverages. Based on this, current studies are mainly focused on the search of new type of yeasts with technological application. Non- Saccharomyces yeasts have always been considered contaminants in the manufacture of wine and beer. Therefore, procedures for eliminating them are routinely utilized such as must pasteurization, addition of sulfite and sanitization of equipment and processing halls. In recent years, the negative perception about non- Saccharomyces yeasts has been changing due to the fact that several studies have shown that during spontaneous fermentations of wine, these yeasts play an important role in the definition of the sensory quality of the final product. Based on this evidence, the fermentative behavior of some non- Saccharomyces yeasts is being studied in deep with the purpose of finding the most adequate conditions and the most suitable strain to be utilized in the production of fermented beverages.

2.1.2. Beer Fermentation

Beer is the most consumed alcoholic beverage worldwide. It is traditionally made from four key ingredients: malted cereals (barley or other), water, hops, and yeast. Each of these ingredients contributes to the final taste and aroma of beer. During fermentation, yeast cells convert cereal-derived sugars into ethanol and CO 2 . At the same time, hundreds of secondary metabolites that influence the aroma and taste of beer are produced. Variation in these metabolites across different yeast strains is what allows yeast to so uniquely influence beer flavor [ 9 ]. Although most breweries use pure yeast cultures for fermentation, spontaneous or mixed fermentation is nowadays used for some specialty beers. These fermentation procedures involve a mix of different yeast species (and bacteria as well) that contribute to the final product sequentially, giving the beer a high degree of complexity. Commonly, breweries have their own stock of selected yeasts for their specific beers. As it is well-known, two types of yeast are used in brewing: S. cerevisiae as the top-fermenting yeast to make ales while S. pastorianus is a bottom-fermenting yeast used in lager brewing processes [ 10 ].

2.1.3. Cider Fermentation

Cider is another alcoholic beverage derived from the apple fruit industry, very popular in different countries in the world, mainly Europe, North America, and Australia [ 11 ]. Although traditional ciders are produced from spontaneous fermentation of juice carried out by autochthonous yeasts, selected S. cerevisiae strains are also commonly used to carry out alcoholic fermentation. This ensures a consistent quality of the finished products [ 12 ]. Some other non- Saccharomyces yeast species are involved in spontaneous fermentation of apple juice for cider production. However, these yeasts contribute at a lesser extent than Saccharomyces and can be producers of off-flavours [ 13 ]. Research articles on this type of product are scarce compared to wine, especially in phenomena associated with microbial activities. The microbiome of wine fermentation and its dynamics, the organoleptic improvement of healthy and pleasant products and the development of starters are now extensively studied. Although the two beverages seem close in terms of microbiome and process (with both alcoholic and malolactic fermentations), the inherent properties of the raw materials and different production and environmental parameters make it worthwhile research on the specificities of apple fermentation. An excellent review of the microbial implications associated with cider production, from ecosystem considerations to associated activities and the influence of process parameters [ 11 ].

In addition to these three worldwide-famous fermented beverages, there are many others made from fruit in various countries in Africa, Asia, and Latin America. Although its consumption is local or regional, in some countries drinks made using fruits such as bananas or grapes as raw materials are very popular. The most widespread alcoholic fruit drink in Eastern Africa is banana beer, which in addition to gastronomic interest is especially culturally relevant. Banana beer is a mixed beverage made from bananas and a cereal flour (often sorghum flour) [ 14 ]. Dates in North Africa, pineapples and cashew fruits in Latin America and jack fruits in Asia are other of the most relevant products.

2.2. Non-Alcoholic Fermentations

Moreover, yeast can act in the fermentation of global non-alcoholic products (bread, chocolate or coffee, beverages such as kefir, sodas, lemonades, and vinegar or even biofuels and other chemicals.

2.2.1. Bread Fermentation

The fermentation of the dough made by the yeasts is the most critical phase in the making of bread. The fermentative yield of yeast cells during this fermentation is crucial and determines the final quality of the bread. Yeasts not only produce CO 2 and other metabolites that influence the final appearance of the dough, volume, and texture, and of course, the taste of the bread. The yeast strain, pregrowth conditions, its activity during the dough fermentation process, the fermentation conditions, as well as the dough ingredients are basic to control the process. The fermentation rate is also conditioned by the ingredients of the dough, including the amounts of sugar and salt used in its preparation. Commercial bread producers currently produce various types of dough such as lean, sweet or frozen dough. Depending on the type of dough, and to obtain optimal fermentation rates, it is recommended to use suitable yeast strains with specific phenotypic traits [ 15 ].

2.2.2. Coffee Fermentation

Yeasts play an important role in coffee production, in the post-harvest phase. Its performance can be done in two phases. On the one hand, aerobically, in which the berries just collected are deposited in a tank and the yeasts are allowed to act. This process is carried out under control of basic parameters, such as time and temperature. Alternatively, coffee berries are deposited in a container mixed with water and microorganisms are allowed to act anaerobically (in the absence of oxygen). This second process is more homogeneous and easy to control than the aerobic. Sometimes, coffee beans are even fermented in a mixed process, first in an aerobic and finally anaerobic manner [ 16 ]. To develop these processes in a satisfactory manner, and to preserve/improve the organoleptic properties of coffee, refine its sweetness, control acidity, give them body or add sensory notes (chocolate, caramel, fruits) mucilage should be removed. The process is naturally carried out by the yeasts present in the mixture, although the process can be improved by the addition of appropriate enzymes (polygalacturonase, pectin lyase, pectin methylesterase) [ 17 ].

2.2.3. Chocolate Fermentation

Raw cacao beans have a bitter and astringent taste, because of high phenolic content. Anthocyanins are one group of these polyphenols, and it both contributes to astringency and provide the reddish-purple color. Fermentation allows the enzymatic breakdown of proteins and carbohydrates inside the bean, creating flavor development. This is aided by microbial fermentation, which create the perfect environment through the fermentation of the cacao pulp surrounding the beans. This processing step enables the extraction of flavor from cacao and contributes to the final acidity of the final product. Yeasts (and also bacteria) ferment the juicy pulp among the cacao beans by different methods, generally following a an anaerobic phase and an aerobic phase. During the anaerobic phase, the sugars of the pulp (sucrose, glucose, fructose) are consumed by yeasts using anaerobic respiration to yield carbon dioxide, ethanol, and low amounts of energy [ 18 , 19 ]. The aerobic stage is dominated by lactic and acetic-acid-producing bacteria [ 20 ].

2.3. Not Only Food: Biofuels and Other Chemicals

The fermentation processes of substrates such as xylose are also of high interest on an industrial level. In addition to expanding the range of substrates that can be used for this purpose, they allow the environmental cost of efficient production of biofuels and other advanced chemicals to be reduced. Some interesting approaches have been made in biorefinery to reprogram yeast for use in these bioprocesses [ 21 , 22 , 23 ].

3. Special Issue on “Yeast Fermentation”

This issue in Microorganisms aims to contribute to the update of knowledge regarding yeasts, regarding both basic and also applied aspects. Among the great contributions to this issue we have a manuscript devoted to the brewing industry and the recent isolation of the yeast Saccharomyces eubayanus [ 24 ]. The use of headspace solid-phase microextraction followed by gas chromatography-mass spectrometry (HS-SPME-GC-MS) has contributed to the production of volatile compounds in wild strains and to compare them to a commercial yeast. All these findings highlight the potentiality of this yeast to produce new varieties of beers. Haile et al. [ 17 ] have explored the possibility to identify and select pectinolytic yeasts that have potential use as a starter culture for coffee fermentation. Almost 30 isolates, eight of them with the ability to produce pectinase enzymes were identified and confirmed by using molecular biology techniques. A helpful bioinformatics tool (MEGA 6) was also used to generate phylogenetic trees able to determine the evolutionary relationship of yeasts obtained from their experiments. Biofuel production by recombinant Saccharomyces cerevisiae strains with essential genes and metabolic networks for xylose metabolism has been also reported [ 23 ]. The authors have shown that the deletion of cAMP phosphodiesterase genes PDE1 and PDE2 can increase xylose utilization. Moreover, the door is opened to provide new targets for engineering other xylose-fermenting strains. The utilization of xylose, the second most abundant sugar component in the hydrolysates of lignocellulosic materials, is a relevant issue. Understanding the relationship between xylose and the metabolic regulatory systems in yeasts is a crucial aspects where hexokinase 2 (Hxk2p) is involved [ 25 ]. All of these processes can be damaged if contaminated. Because most fermentation substrates are not sterile, contamination is always a factor to consider. With a very interesting approach, a genetically modified strain of Komagataella phaffii yeast was used for the use of glycerol as a base substance in lactate production. Polyactide, a bioplastic widely used in the pharmaceutical, automotive, packaging and food industries was produced. The disruption of the gene encoding arabitol dehydrogenase (ArDH) was achieved, which improves the production of lactic acid by K. phaffii as a biocatalyst [ 26 ]. Seo et al. [ 27 ] have developed and proposed alternative solutions to control contamination. This review includes information on industrial uses of yeast fermentation, microbial contamination and its effects on yeast fermentations. Finally, they describe strategies for controlling microbial contamination.

Acknowledgments

Thanks to all the authors and reviewers for their excellent contributions to this Special Issue. Additional thanks to the Microorganisms Editorial Office for their professional assistance and continuous support.

Conflicts of Interest

The editors declares no conflict of interest.

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Yeast and the expansion of bread dough

In association with Nuffield Foundation

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Try this class practical to investigate how temperature affects yeast and the expansion of bread dough

Yeast is a microbe used in bread making which feeds on sugar. Enzymes in yeast ferment sugar forming carbon dioxide and ethanol. The carbon dioxide makes the bread rise, while the ethanol evaporates when the bread is baked. In this experiment, students investigate the effect of different temperatures on yeast activity and the expansion of the bread dough.

It is best if each group does the activity at one temperature and then shares the results with other groups.

• Spatula or glass rod
• Beaker, 100 cm 3
• Measuring cylinder, 250 cm 3
• Measuring cylinder, 50 cm 3
• Thermometer, 0–100 °C
• Graph paper
• Access to a balance (1 d.p.)
• Access to water baths set at 20 °C, 30 °C and 37 °C (see note 2 below)
• Plain flour, 25 g
• Yeast suspension, 30 cm 3 (see note 3 below)

Health, safety and technical notes

• Read our standard health and safety guidance.
• If thermostatically controlled water baths are not available then large beakers of water maintained at the three temperatures can be used. The water temperature in each beaker needs to be monitored using a thermometer, and access to a supply of hot water, eg from a kettle, is needed to top up the beaker as the temperature falls. 
• The yeast suspension is made by stirring 7 g of dried yeast with 450 cm 3 of warm water. If fresh yeast is used it should not have been kept too long. Fresh yeast can be kept in the freezer for up to two months.
• Add 25 g of flour to a beaker and then add 1 g of sugar.

Source: Royal Society of Chemistry

Weigh flour and sugar for a simple bread dough

• Take 30 cm 3 of the yeast suspension in a 50 cm 3 measuring cylinder. Add the yeast suspension to the flour and sugar. Stir with a spatula or glass rod until a smooth paste, which can be poured, has been obtained.
• Pour the paste into a 250 cm 3 measuring cylinder. Take great care not to let the paste touch the sides – this is very important.

Make a paste from the flour, sugar and yeast suspension and pour it into a measuring cylinder

• Note the volume of paste in the cylinder. Place the cylinder in one of the water baths. Record the temperature and note the volume of paste every two minutes for about 30 minutes. A results table is useful here.
• Plot a graph to show how the volume of the dough increases with time. Plotting the results from groups, with the water baths at different temperatures, on the same graph will allow comparison of results.

Teaching notes

It is important that the paste does not touch the sides of the measuring cylinder when the students pour it from the beaker – this is easier said than done. One way of achieving this is to use a large plastic funnel which has had the stem cut off to leave a hole large enough for the paste to flow through the funnel. Alternatively, a suitable plastic drinks bottle could be cut to produce a wide mouthed ‘funnel’.

After 35–45 minutes the protein (gluten) breaks and carbon dioxide gas escapes.

Possible extensions for this activity could be to investigate the effect of substrate concentration (sugar), enzyme concentration and/or pH value on enzyme reactions.

Additional information

This is a resource from the  Practical Chemistry project , developed by the Nuffield Foundation and the Royal Society of Chemistry.

Practical Chemistry activities accompany  Practical Physics  and  Practical Biology .

© Nuffield Foundation and the Royal Society of Chemistry

• 11-14 years
• 14-16 years
• Practical experiments
• Biological chemistry

Specification

• 7. Investigate the effect of a number of variables on the rate of chemical reactions including the production of common gases and biochemical reactions.
• 9. Consider chemical reactions in terms of energy, using the terms exothermic, endothermic and activation energy, and use simple energy profile diagrams to illustrate energy changes.
• 2. Develop and use models to describe the nature of matter; demonstrate how they provide a simple way to to account for the conservation of mass, changes of state, physical change, chemical change, mixtures, and their separation.
• Awareness of the contributions of chemistry to society, e.g. provision of pure water, fuels, metals, medicines, detergents, enzymes, dyes, paints, semiconductors, liquid crystals and alternative materials, such as plastics, and synthetic fibres; increasi…
• Enzymes as catalysts produced by living cells (two examples).

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Lab Explained: Production of Yeast Fermentation

• Lab Explained: Production of Yeast…

Introduction

Yeast is commonly known as a baking material for making bread and beer. However, under biological jargon it is a group of eukaryotic, single-celled microorganisms that makes up almost 1% of all fungal species. 1 They are found in soils and on plant surfaces, like flower nectar and fruits, and they reproduce asexually through budding. This is where a small daughter cell grows on the parent cell whose nucleus then duplicates and then separates with the daughter cell. This reproduction can happen at a rate of up to once every 90 minutes. 2 However, some yeast is an exception and reproduces by binary fission, a process in which the DNA duplicates and the cytoplasm splits evenly into two identical daughter cells.

Most importantly, yeast undergoes metabolism without the presence of oxygen, thus respiring anaerobically. The carbon dioxide that is produced is what makes bread rise when using the specific type; Saccharomyces cerevisiae , also known as baker’s yeast.

The breakdown of glucose by yeast:

C6H12O6 → 2C2H5OH + 2CO2

Glucose → ethanol + carbon dioxide

The glucose is broken down by glycolysis. However, there is more to the process than this simple arrow. When respiring anaerobically, yeast performs glycolysis where it converts pyruvate further into ethanol and carbon dioxide. Although the process requires 2 ATP, it produces 4; a net gain of 2 ATP molecules that can be used for energy. Fermentation rate is affected by factors like temperature, saccharide concentration, and type of sugar solution.

Monosaccharides are the monomers 3 of carbohydrates, consisting of glucose, fructose, and galactose. They provide quick, accessible energy that is easily broken down. Disaccharides are made of two monosaccharides, consisting of lactose, maltose, and sucrose. They are held together by a covalent bond 4 through a condensation reaction. 5 Thus, their more complex structures make them harder to break down compared to monosaccharides. These are the sugars and the monomers each sugar is made up of.

Investigations

2.1 hypothesis.

Glucose will produce the largest amount of carbon dioxide, followed, by fructose, sucrose, and lactose. Although they all have similar chemical formulas, they differ in structure and

stereochemistry. 6 The monosaccharides will perform better because they require less energy to be broken down and thus create the product of carbon dioxide at a faster and higher rate.

H : The carbon dioxide produced will be the same for each sugar type.

H 1 : The addition of monosaccharides will produce more carbon dioxide than disaccharides.

2.2 Variables

Independent Variable: Type of saccharide

Dependent Variable: Time of each observation (minutes)

Controlled Variables:

• Beaker size: 250ml
• Keeping this consistent would allow the same space for each reaction to take place.
• Amount of yeast: 2.0g
• Type of yeast: Alnatura Backhefe
• Ingredients: Yeast dried from organic farming
• This was important to monitor because different yeast that has different origins could have different reactions.
• Amount of distilled water: 50ml
• Maintaining a consistent amount of ingredients ensured that the chemical reactions would have fair, equal environments.
• Amount of saccharide: 5.0g

Uncontrolled Variables:

• Room temperature: 21°C
• This was monitored using a thermometer. This was important to keep consistent because increased temperatures speed up reactions. However, the lab is quite large, so it was impossible to control this variable.

2.3 Preliminary Experiment

A preliminary experiment was carried out a day before our designated internal assessment time. I thought it was appropriate to prepare for my actual experiment to ensure that all the materials were working, and my procedure demonstrates efficacy. My method was identical to that of my real one, however, I did make one change. Originally, my saccharide to distilled water ratio was 5g:100ml. This was not fitting for my time frame of 30 minutes. Thus, I changed my measurements and made the solution much more concentrated; 5g:50ml. This increased the speed of the reaction so I could take more readings within a smaller span of time, allowing me to also improve my reliability. 7

In addition to adapting my method, I also conducted a control test. I mixed 2g of yeast with 50ml of distilled water and no carbon dioxide was released. Now that I had a method that worked, I could safely move on to my final internal assessment procedure.

3.1 Apparatus

• Gas Syringe – to measure produced carbon dioxide (±0.05ml)
• Mixer – to create and mix the solutions of a saccharide and distilled water
• Stir Rod (or Flea) – magnet used to mix the solutions
• Stands – to hold apparatus in place
• Scale – to measure necessary materials (± 0.05g)
• Stopper – to prevent any carbon dioxide loss throughout the structure
• Clamps – to hold flask and gas syringe in place
• Graduated cylinder – to measure the amount of each ingredient (±0.1ml)
• Flask – to contain the site of the reaction
• Rubber Tube – to connect the glass pipes
• Baking Yeast (Backhafer)
• Glucose, Fructose, Sucrose, Lactose
• Distilled Water – to create solution that is added to yeast

3.2 Methodology

• Pour 2g of yeast into the flask
• Add 50ml distilled water
• Put mixer on 500 rpm
• Record results at both 15 and 30
• The mixture should not release any carbon dioxide, as there is no respiration.

Saccharide Experiment:

• Mix 5g of the saccharide with 50ml distilled water in a beaker until clear
• Make sure the valve is open so the carbon dioxide can flow through to the gas syringe
• Pour the saccharide and distilled water solution into the flask
• Immediately close the flask with the stopper
• Immediately start timer
• Record results at 15 minutes (from gas syringe)
• Stop timer at 30 minutes and record results
• Clean apparatus between uses
• Repeat 4 times per sugar for each sugar

3.3 Justification

All of my measurements for my materials were done with a digital scale where I could control how much of each ingredient I was using down to the nearest milligram. I additionally used two of them, in case one were to malfunction or give a different result. Therefore, I know that I did not have any false readings or amounts or yeast, sugars, or distilled water. Unless, of course, both digital apparatuses malfunctioned.

I repeated my method four times with each sugar to increase reliability and make sure I was getting consistent results. Additionally, when a trial went wrong in any way, I would clean up and re-do it. For example, one time I realized that I had labelled my flask incorrectly and lost track of which sugar I had put in the solution. I properly disposed of it, cleaned the materials, and started over to ensure that I was being accurate, and no leftover waste or excess ingredients would affect the results i.e. each experiment would be independent from one another.

3.4 Risk Assessment

Safety issues: None of the materials I used were harmful or put me at risk in any way.

Ethical issues: Nothing in my internal assessment could be deemed unethical in any way. Environmental issues: The disposal of yeast in a drainage or water system can result in adecrease of oxygen for anything that is further down the pipes. This oxygen shortage can throw off the balance of any ecosystems associated with the drain in question. Thus, I diluted the yeast mixture with water and then disposed of it in the waste container.

Raw and Processed Data

4.1 qualitative observations.

• The saccharide mixture combined with the yeast started to create a layer of froth/bubbles at the top.
• The mixture previously mentioned also left condensation all over the inside of the flask.
• Some sugars were harder to dissolve in the distilled water than others, namely the fructose. This is most likely due to the more complex structure that makes it harder to mingle with the H2O. The more monomers and bonds within a compound, the more difficult it is for any other molecule, in this case water, to interfere and dissolve it.

4.2 Raw Data

4.3 Processed Data

From this graph, we can very well see that some sugars simply performed better, specifically glucose and fructose. The disaccharides – fructose and lactose – had a much lower fermentation rate. Thus, my hypothesis is being supported.

Had I not discarded the anomalies present in my raw data for sucrose, the standard deviations would have been over 5x as large. Because standard deviation is the value describing how much the data differs from the mean, it is very evident that such a large change in the value all due to one anomaly was unnecessary in my final conclusions and analysis of data. It would have negatively impacted the scope of my investigation because it is very obviously due to an error in my method (see weaknesses).

5.1 Conclusion

Looking at table 3 and graph 1, the hierarchy of efficacy in producing carbon dioxide reads as follows; glucose, fructose, sucrose, and lactose. When speaking in terms of standard deviation, sucrose and lactose had the smallest. Table 4 further supports this. Thus, sucrose and lactose were the most consistent and differed the least from their mean. This could be due to the substances being stored correctly or never having been disrupted since their extraction from the lab they were bought from. From table 4, we can further infer that since the F value is greater than the F crit value, the values are not at all equal, rejecting my null hypothesis.

A higher standard deviation, as seen in fructose, could be a result of improper care of the sugars themselves. For example, placing them in warm temperatures or letting them be vulnerable to sunlight could cause them to melt or change in structure and thus alter from molecule to molecule.

Regarding other academic investigations of yeast, my investigation gave results that were expected by other biologists. To quote from the Journal of Undergraduate Biology Laboratory Investigations, “The monosaccharides we used in our experiments (glucose and honey)produced a higher rate of CO2 than the disaccharides (refined sucrose and lactose) did.” This directly aligns with my alternative hypothesis and experiment results.

From my results, it can be concluded that yeast is affected differently depending on the type of saccharide it encounters. Thus, it supports my hypothesis: Glucose will produce the largest

amount of carbon dioxide, followed, by fructose, sucrose, and lactose. As mentioned before, their chemical formulas being almost identical has little to do with it. It is all in the structure. The monosaccharides require less energy to be broken down and thus create the product of carbon dioxide. This is proven by the fact that every cell has the ability to break down glucose, whereas only the liver can break down fructose. 9 Even then, when breaking down fructose, it is first converted into glucose where then glycolysis can take place.

5.2 Improvements

The apparatus I used was not optimal. For example, my mixer did not have a digital display of how fast my rpm was. Thus, I could have been mixing the yeast and sugar solutions at different speeds for each trial. The speed could have affected the rate of the reaction due to how well the molecules were interacting and colliding with each other, and thus affecting the production of carbon dioxide.

In addition, I used quite a large flask compared to the 50ml of solution I had to use. It was made for 250ml, so it is imaginable how much extra space was within the container. This may have decreased the accuracy of the measurements I took from the gas syringe, because there was also gas trapped in the flask itself. Perhaps I could have additionally measured the volume of the container that did not have the solution (the gap between the liquid surface and the gas syringe) and added that to my volumetric values of carbon dioxide to increase my accuracy.

Finally, the yeast packages I used were all from the same store, however it is possible that they were sourced from different farms. Therefore, the structure of the yeast molecules used could have differed due to different methods used while they were farmed. Albeit, this would be a very difficult, arbitrary thing to track, and is a stretch in affecting the results of my assessment.

What my investigation did do extremely well was independence. It is extremely unlikely that and ingredients unintentionally mixed or met due to my rigorous cleaning and replacing of the apparatus. In addition, my method of using the mixer was helpful because it ensured the solutions were extremely even and constant throughout (e.g., no leftover sugar at the bottom).

5.3 Extensions

To extend the current scope of my study, I would be enthusiastic to try and see how different types of yeast would affect my results. In this internal assessment, I only used brewers/baker’s yeast that is simply dried from a farm. However, there are hundreds of other kinds found in a variety of different places. I would measure how these saccharides, and possibly also more, e.g. maltose, affect the fermentation rate of yeasts like torula (used to create paper) and fission yeast (alternative to brewing yeast) or fresh yeast.

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Yeast-Air Balloons

The purpose of any leavener is to produce the gas that makes bread rise. Yeast does this by feeding on the sugars in flour, and expelling carbon dioxide in the process.

While there are about 160 known species of yeast, Saccharomyces cerevisiae, commonly known as baker's yeast, is the one most often used in the kitchen. Yeast is tiny: Just one gram holds about 25 billion cells. That amount of fungi can churn out a significant amount of carbon dioxide, provided it has the simple sugars it uses as food. Fortunately, yeast can use its own enzymes to break down more complex sugars—like the granulated sugar in the activity below—into a form that it can consume.

Make a yeast-air balloon to get a better idea of what yeast can do.

Did You Know?

What do i need.

1 packet of active dry yeast

1 cup very warm water (105° F-115° F)

2 tablespoons sugar

a large rubber balloon

a small (1-pint to 1-liter) empty water bottle

Kids, please don t try this at home without the help of an adult.

What do I do

Stretch out the balloon by blowing it up repeatedly, and then lay it aside.

Add the packet of yeast and the sugar to the cup of warm water and stir.

Once the yeast and sugar have dissolved, pour the mixture into the bottle. You ll notice the water bubbling as the yeast produces carbon dioxide.

Attach the balloon to the mouth of the bottle, and set both aside.

Step 5: After several minutes, you ll notice the balloon standing upright. If you don t see anything happen, keep waiting. Eventually, the balloon will inflate.

What's going on.

As the yeast feeds on the sugar, it produces carbon dioxide. With no place to go but up, this gas slowly fills the balloon.

A very similar process happens as bread rises. Carbon dioxide from yeast fills thousands of balloonlike bubbles in the dough. Once the bread has baked, this is what gives the loaf its airy texture.

What Else Can I Try?

Try the same experiment, but this time use about a tablespoon of baking powder instead of yeast, and leave out the sugar. What differences do you notice? Which leavener takes longer to fill up the balloon?

Also, try the same experiment using hotter and colder water. Use a thermometer to measure the temperature of the water. At what temperature is the yeast most active? At what temperatures is it unable to blow up the balloon?

Remember Me

Shop Experiment Sugar Fermentation by Yeast Experiments​

Sugar fermentation by yeast.

Experiment #24 from Investigating Chemistry through Inquiry

Introduction

Yeast can metabolize sugar in two ways, aerobically , with the aid of oxygen, or anaerobically , without oxygen. When yeast metabolizes a sugar under anaerobic conditions, ethanol (CH 3 CH 2 OH) and carbon dioxide (CO 2 ) gas are produced. An equation for the fermentation of the simple sugar glucose (C 6 H 12 O 6 ) is:

The metabolic activity of yeast can be determined by the measurement of gas pressure inside the fermentation vessel.

In the Preliminary Activity, you will use a Gas Pressure Sensor to monitor the pressure inside a test tube as yeast metabolizes glucose anaerobically. When data collection is complete, you will perform a linear fit on the resultant graph to determine the fermentation rate.

After completing the Preliminary Activity, you will first use reference sources to find out more about sugar fermentation by yeast before you choose and investigate a researchable question dealing with fermentation.

Sensors and Equipment

This experiment features the following sensors and equipment. Additional equipment may be required.

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This experiment is #24 of Investigating Chemistry through Inquiry . The experiment in the book includes student instructions as well as instructor information for set up, helpful hints, and sample graphs and data.

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Biology Experiments on the Fermentation of Yeast

What Are Some Common Uses of Yeast?

Yeast is a fungal microorganism that man has usedsince before he had a written word. Even to this day, it remains a common component of modern beer and bread manufacture. Because it is a simple organism capable of rapid reproduction and even faster metabolism, yeast is an ideal candidate for simple biology science experiments that involve the study of fermentation.

What is Fermentation?

Fermentation is the biological process by which yeast consumes simple sugars and releases alcohol and carbon dioxide. For the most part, fermentation requires a mostly aquatic environment to occur. Different yeasts respond differently to changes in environment, making some better for baking and others for brewing. Bakers use fermentation to add CO2 bubbles to bread dough. During baking, these bubbles make the bread light and fluffy while the alcohol boils away. Brewers take care to preserve the alcohol of fermentation and use the CO2 to help build a frothy head for their potent beverages.

Indirect Life Test Experiments

The first experiment that should come to mind when examining yeast is determining whether or not yeast is a living organism. While it would be easy to rely on foreknowledge about the nature of yeast, more is learned by application of scientific method. If yeast is alive, it should consume food, respire and reproduce. Indirect tests look for clues that these processes are taking place. For such experiments, you should measure the amount of CO2 released by yeast that are digesting sugar water in test tubes with balloons attached. Use Benedict's solution to test for the presence of sugar in the final product.

Salinity Experiments

Fermentation is a delicate process that relies on ideal conditions to occur. Experiments that study how it responds to salinity are of particular interest to science and industry alike. Your project can either take a single type of yeast and vary the amount of salt in the solution to see if there is an ideal salinity, or alternately, use various yeasts to see how they respond to the same level of salt. In the latter experiment, make sure to use yeasts from many industries, since most baker's yeasts fare poorly in saline conditions.

Sugar Experiments

While it's clear that yeast requires sugar for fermentation, there are many different sugars that yeast could use for fuel. You can perform a number of experiments to determine which ones promote the highest level of yeast growth. In one, you can add yeast to various beverages, such as fruit juices and non-carbonated sports drinks to see which environment produces the most CO2. Another can use various sweeteners such as granulated sugars, syrups and nectars (such as agave) placed in weak solutions. You can measure CO2 production with balloons placed over the reacting test tubes, or simply observe the bubbles produced and make a relative comparison.

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About the Author

Andy Klaus started his writing career contributing science and fiction articles to Dickinson High School's newsletters back in 1984. Since then, he has authored novels and written technical books for health-care companies such as VersaSuite. He has covered topics varying from aerospace to zoology and received an associate degree in science from College of the Mainland.

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Identification of Yeast Strain YA176 for Bio-Purification of Soy Molasses to Produce Raffinose Family Oligosaccharides and Optimization of Fermentation Conditions

• Original Article
• Published: 28 September 2024

Cite this article

• Zhilei Fu 1 , 2 , 3 ,
• Shuang Cheng 1 ,
• Jinghao Ma 3 ,
• Rana Abdul Basit 3 ,
• Yihua Du 3 ,
• Shubin Tian 4 &
• Guangsen Fan   ORCID: orcid.org/0000-0001-5758-130X 1 , 3 , 4  

Soybean molasses, which contains high levels of raffinose family oligosaccharides (RFOs) such as stachyose and raffinose, is subjected to a process of bio-purification to remove sucrose while maintaining the RFOs, consequently increasing its value. This study employed morphological observation, physiological and biochemical studies, and molecular biology techniques to identify YA176, a yeast strain renowned for its effective bio-purification of soy molasses. Through single-factor and orthogonal experiments, optimal bio-purification conditions were established. YA176, belonging to Wickerhamomyces anomalus , demonstrated robust growth across a wide range of temperature and pH levels, coupled with remarkable tolerance to glucose, sucrose, and NaCl up to 41.2%, 47.3%, and 10%, respectively. Under these optimized conditions, YA176 efficiently utilized sucrose while preserving 93.3% of raffinose and 78.6% of stachyose, ensuring the retention of functional RFOs. In summary, yeast strain YA176 exhibits exceptional bio-purification abilities, making it an ideal candidate for producing functional RFOs from soy molasses.

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Valorization of Soybean Molasses as Fermentation Substrate for the Production of Microbial Exocellular β-Glucan

Potential Hexose Fermenting Yeast for Conversion of Sugary and Starchy Raw Materials into Ethanol

Bioethanol from Soybean Molasses

Data availability.

All generated or analyzed data as well as the utilized software and materials have been included in the published article. The generated results during the study are available from the corresponding authors upon reasonable request.

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This research was supported by the Open Research Fund Program of Henan Key Laboratory of Industrial Microbial Resources and Fermentation Technology (HIMFT20190205), Doctoral Research Initiation Fund, Hebei Normal University for Nationalities (DR2022004), and Beijing Natural Science Foundation (6222003 and 6164029).

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Zhilei Fu, Shuang Cheng & Guangsen Fan

School of Biology and Food Science, Hebei MinZu Normal University, Chengde, 067000, China

Key Laboratory of Geriatric Nutrition and Health (Beijing Technology and Business University), Ministry of Education, No. 11, Fucheng Road, Haidian District, Beijing, 100048, China

Zhilei Fu, Jinghao Ma, Rana Abdul Basit, Yihua Du & Guangsen Fan

Sweet Code Nutrition and Health Institute, Zibo, 256306, China

Shubin Tian & Guangsen Fan

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Zhilei Fu and Yihua Du have performed the experimental analysis study. Jinghao Ma and Shubin Tian have carried out the analysis and interpretation of the results. Zhilei Fu wrote the manuscript. Shuang Cheng and Guangsen Fan have designed the study concept and interpreted the data. Rana Abdul Basit read and revised the manuscript and approved the final version. All authors read and approved the final manuscript.

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Correspondence to Guangsen Fan .

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Fu, Z., Cheng, S., Ma, J. et al. Identification of Yeast Strain YA176 for Bio-Purification of Soy Molasses to Produce Raffinose Family Oligosaccharides and Optimization of Fermentation Conditions. Appl Biochem Biotechnol (2024). https://doi.org/10.1007/s12010-024-05065-4

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Accepted : 19 September 2024

Published : 28 September 2024

DOI : https://doi.org/10.1007/s12010-024-05065-4

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Effects of a liquid and dry Saccharomyces cerevisiae fermentation product feeding program on ruminal fermentation, total tract digestibility, and plasma metabolome of Holstein steers receiving a grain-based diet

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Oluwaseun A Odunfa, Anjan Dhungana, Zhengyan Huang, Ilkyu Yoon, Yun Jiang, Effects of a liquid and dry Saccharomyces cerevisiae fermentation product feeding program on ruminal fermentation, total tract digestibility, and plasma metabolome of Holstein steers receiving a grain-based diet, Journal of Animal Science , Volume 102, 2024, skae223, https://doi.org/10.1093/jas/skae223

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The study aimed to determine the effects of a postbiotic feeding program consisting of liquid and dry Saccharomyces cerevisiae fermentation product ( SCFP ) on ruminal fermentation, digestibility, and plasma metabolome of Holstein steers receiving a grain-based diet. Eight Holstein steers (body weight, BW , 467 ± 13.9 kg) equipped with rumen cannulas were used in a crossover design study, with 21 d per period and a 7-d washout period in between periods. Steers were stratified by initial BW and assigned to 1 of 2 treatments. The treatments were 1) Control, basal finishing diet only (CON); 2) SCFP, 1-d feeding of liquid SCFP (infused into the rumen via the cannula at 11 mL/100 kg BW) followed by daily feeding of dry SCFP (12 g/d, top-dressed). Feed and spot fecal samples were collected during days 17 to 20 for determination of digestibility and fecal excretion of N, P, Cu, and Zn. Digestibility was measured using acid-insoluble ash as an internal marker. Blood samples were collected on day 21 before the morning feeding. Rumen fluid samples were collected on days 0, 1, 2, 3, 5, and 21 via rumen cannula. Results were analyzed with the GLIMMIX procedure of SAS 9.4 (SAS, 2023). Treatment did not affect dry matter intake ( P  = 0.15) and digestibility ( P  ≥ 0.62). The fecal output and absorption of Zn, Cu, P, and N were not affected ( P  > 0.22) by treatment. On day 1, the liquid SCFP supplementation tended to reduce ( P  = 0.07) ruminal VFA concentration and increased ( P  < 0.01) the molar proportion of valerate. Feeding SCFP tended to increase total ruminal VFA on day 5 ( P  = 0.08) and significantly increased total VFA on day 21 ( P  = 0.05). Ruminal NH 3 –N was reduced ( P  = 0.02) on day 21 by supplementing SCFP. Treatment did not affect the production of proinflammatory cytokines, interleukin ( IL )-1β ( P  > 0.19), and IL-6 ( P  > 0.12) in the whole blood in response to various toll-like receptor stimulants in vitro. Feeding SCFP enriched ( P  ≤ 0.05) plasma metabolic pathways, including citric acid cycle, pyrimidine metabolism, glycolysis/gluconeogenesis, retinol metabolism, and inositol phosphate metabolism pathways. In summary, supplementing liquid SCFP with subsequent dry SCFP enhanced ruminal total VFA production and reduced NH 3 –N concentration in the rumen. Furthermore, feeding SCFP enriched several important pathways in lipid, protein, and glucose metabolism, which may improve feed efficiency of energy and protein in Holstein steers.

Lay Summary

Previous research has shown the positive effects of Saccharomyces cerevisiae fermentation product ( SCFP ) on beef cattle performance. Liquid SCFP is a novel form of SCFP and has the potential to prime the rumen environment and improve subsequent ruminal fermentation and performance of Holstein steers receiving a grain-based diet. We investigated the impact of a novel feeding program using liquid and dry SCFP on ruminal fermentation, digestibility, and plasma metabolome of beef steers. Compared to non-supplemented control, feeding SCFP did not affect nutrient digestibility but enhanced ruminal fermentation, as shown by improved total volatile fatty acid production by rumen microbes after 5 d of supplementation. The supplementation of SCFP also enriched several plasma metabolic pathways related to energy and nitrogen metabolism, such as the citric acid cycle, pyrimidine metabolism, and glycolysis/gluconeogenesis pathways.

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Sustainable exploitation of wine lees as yeast extract supplement for application in food industry and its effect on the growth and fermentative ability of lactiplantibacillus plantarum and saccharomyces cerevisiae.

1. Introduction

2. materials and methods, 2.1. materials, 2.2. production of wine lees yeast extract powder, 2.3. inoculum preparation, 2.4. test culture media, growth monitoring and cell enumeration, 2.5. data analysis, 2.6. evaluation of fermentative ability, 2.7. determination of ph, reducing sugars and ethanol content in fermentation media, 2.8. statistical analysis, 3. results and discussion, 3.1. growth kinetics of lactiplantibacillus plantarum 2035, 3.2. growth kinetics of saccharomyces cerevisiae ncyc187, 3.3. fermentative ability, 4. conclusions, supplementary materials, author contributions, institutional review board statement, informed consent statement, data availability statement, conflicts of interest.

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Kokkinomagoulos, E.; Kandylis, P. Sustainable Exploitation of Wine Lees as Yeast Extract Supplement for Application in Food Industry and Its Effect on the Growth and Fermentative Ability of Lactiplantibacillus plantarum and Saccharomyces cerevisiae . Sustainability 2024 , 16 , 8449. https://doi.org/10.3390/su16198449

Kokkinomagoulos E, Kandylis P. Sustainable Exploitation of Wine Lees as Yeast Extract Supplement for Application in Food Industry and Its Effect on the Growth and Fermentative Ability of Lactiplantibacillus plantarum and Saccharomyces cerevisiae . Sustainability . 2024; 16(19):8449. https://doi.org/10.3390/su16198449

Kokkinomagoulos, Evangelos, and Panagiotis Kandylis. 2024. "Sustainable Exploitation of Wine Lees as Yeast Extract Supplement for Application in Food Industry and Its Effect on the Growth and Fermentative Ability of Lactiplantibacillus plantarum and Saccharomyces cerevisiae " Sustainability 16, no. 19: 8449. https://doi.org/10.3390/su16198449

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