Saccharomyces cerevisiae in bakery industries

Name : Rinanti Pritanova ID Number : 1-4209-026 Food Technology 4 Life Sciences Swiss German University

1.

Introduction

The aim of this paper is to discuss about Saccharomyces cerevisiae, both about its characteristic and also about its production process in food industries. Actually, there are many kind of purpose in using Saccharomyces cerevisae in food industries, but this time the common used in bread making that will be explained. The fermentations focus on a maximum biomass yield with limited ethanol production, favouring the oxidative or aerobic metabolic yeast pathway. Bakers usually use yeast as a leavening agent in the rising of dough for baking. A secondary contribution of yeast to bread is flavouring and aroma.

2.

Cell Overview

2.1 Name
The binomial name of this micro bacterium is Saccharomyces cerevisiae. "Saccharomyces" derives from Latinized Greek and means "sugar mold" or "sugar fungus", saccharo- being the combining form "sugar-" and myces being "fungus". Cerevisiae comes from Latin and means "of beer".Other names for the organism are :

S. cerevisiae (short form of the scientific name) Brewer's yeast, (though other species are also used in brewing) Ale yeast Top-fermenting yeast Baker's yeast Budding yeast

Figure 2.1 S. cerevisiae under DIC microscopy

The scientific classification for Saccharomyces cerevisiae is : Kingdom : Fungi Phylum : Ascomycota Subphylum : Saccharomycotina Class : Saccharomycetes Order : Saccharomycetales Family : Saccharomycetaceae Genus : Saccharomyces Species : S. cerevisiae

2.2 Type
Saccharomyces cerevisiae is a eukaryotic organism. It is usually spherical, oval or cylindrical in shape and a single cell of S. cerevisiae is around 8m in diameter, around 10 m long by 5 m wide. Each cell has a double-layered wall (thick-walled), which is permeable to certain substances and in this way food material is taken into the cell and metabolites leave it. It reproduces by a division process known as budding, or in some cases by division (fission). It is one of the most intensively studied eukaryotic model organisms in molecular and cell biology, much like Escherichia coli as the model bacteria.

Figure 2.2 Saccharomyces cerevisiae. Numbered ticks are 10 micrometres apart

2.3 Usage
As it was said before, S. cerevisiae is most useful yeast owing to its use since ancient times in baking and brewing. It is the microorganism behind the most common type of fermentation. This species is also the main source of nutritional yeast and yeast extract.

2.4 Isolation
The yeast is believed that it was originally isolated from the skins of grapes (one can see the yeast as a component of the thin white film on the skins of some dark-colored fruits such as plums; it exists among the waxes of the cuticle).

2.5 Life Cycle

There are two forms in which yeast cells can survive and grow: haploid and diploid. The haploid cells undergo a simple life cycle of mitosis and growth, and under conditions of high stress will, in general, die. The diploid cells (the preferential 'form' of yeast) similarly undergo a simple life cycle of mitosis and growth, but under conditions of stress can undergo sporulation, entering meiosis and producing a variety of haploid spores, which can proceed on to mate.

2.6 Cell Cycle

Growth in yeast is synchronised with the growth of the bud, which reaches the size of the mature cell by the time it separates from the parent cell. In rapidly growing yeast cultures, all the cells can be seen to have buds, since bud formation occupies the whole cell cycle. Both mother and daughter cells can initiate bud formation before cell separation has occurred. In yeast cultures growing more slowly, cells lacking buds can be seen, and bud formation only occupies a part of the cell cycle. The cell cycle in yeast normally consists of the following stagesG1, S, G2, and M which are the normal stages of mitosis. Although most yeasts reproduce only as single cells, under some conditions some yeasts can form filaments.

3. Production Overview
3.1 Substrate Used
S. Cerevisiae primarily uses molasses as its growth substrate the main constituent of which is the disaccharide sucrose. Sucrose is composed of the two monosaccharides glucose and fructose. Glucose degradation can proceed via two distinct pathways, depending on the availability of oxygen within the

system. Fructose is degraded in much the same way, differing in the initials reactions for its utilization due to its different conformation. In the presence of oxygen, the oxidative pathway is followed, producing biomass at the expense of glucose breakdown. As oxygen is consumed to support the degradation of glucose, a critical point arises where the reduced availability of oxygen forces the cell to commence anaerobic metabolism as its primary means of growth. This second pathway is known as the reductive pathway and results in the production of biomass along with ethanol (C2H6O). Furthermore, in the presence of oxygen, ethanol can be utilized as a substrate in the oxido-reductive pathway to produce further quantities of biomass. However, bakers yeast production simply requires the production of biomass, therefore the additional production of ethanol is undesired and so the system is designed as to purposely direct cells into the oxidative pathway, producing only biomass. For this reason, methods have been studied to optimise the growth of S. Cerevisiae on glucose by regulating the oxygen uptake and glucose consumption by the yeast cell, consequently regulating which growth pathway the cell will enter.6 The findings of such research concluded that in bakers yeast fermentation, the use of fedbatch fermentation allows the constant supply of sugar, such that sugar substrate will not accumulate but instead accommodate the constant growth of S. cerevisiae i.e. the sugar is utilized as it is added making the process more efficient. Also constant oxygen sparging within the reactor allows the emergence of the oxidative pathway as the sole route of yeast growth.

3.2 Growth Condition

As a living organism yeast primarily requires sugars, water and warmth to stay alive. In addition, albumen or nitrogenous material is also necessary for yeast to thrive.Yeasts flourish in habitats where sugars are present, such as fruits, flowers and bark of trees. However, commercial yeasts of today are quite different from wild strains due to genetic manipulation, allowing them to grow in previously unsuitable conditions. All strains of S. cerevisiae can grow aerobically on glucose, maltose, and trehalose and fail to grow on lactose and cellobiose. However, growth on other sugars is variable. Galactose and fructose are shown to be two of the best fermenting sugars. The ability of yeasts to use different sugars can differ depending on whether they are grown aerobically or anaerobically. Some strains cannot grow anaerobically on sucrose and trehalose. All strains can use ammonia and urea as the

sole nitrogen source, but cannot use nitrate, since they lack the ability to reduce them

to ammonium ions. They can also use most amino acids, small peptides, and nitrogen bases as a nitrogen source. Histidine, glycine, cystine, and lysine are, however, not readily used. S. cerevisiae does not excrete proteases, so extracellular protein cannot be metabolized. Yeasts also have a requirement for phosphorus, which is assimilated as a dihydrogen phosphate ion, and sulfur, which can be assimilated as a sulfate ion or as organic sulfur compounds such as the amino acids methionine and cysteine. Some metals, like magnesium, iron, calcium, and zinc, are also required for good growth of the yeast. The yeast culture is grown up to a stage where it is large enough to start the process in the factory. Thousands of litres of yeast are produced in specially designed fermenters, fed with a molasses broth. When growth is complete the fermented yeast is separated from the medium in which it was grown, resulting in a creamy suspension of concentrated active yeast. Molasses is a by-product of sugar refining and contains about 50% sugar.16 It serves as the source of carbon and energy for the process. It is supplemented with a number of nitrogenous compounds and vitamins such as biotin16, which are required for the proper and efficient growth of the yeast cells.

3.3 Operating Mode

In bread making, the flour used contains starch, protein and an enzyme amylase. The flour is mixed with water to form dough. Amylase digests the starch into glucose. Lack of O2 inside the dough causes the yeast to respire anaerobically. As a result glucose is fermented to alcohol and CO2 the CO produced causes the dough to rise and because of this, cavities appear. Alcohol produced evaporates during baking. One yeast cell can ferment approximately its own weight of glucose per hour, giving rise to large volumes of CO2.

The enzymes which are formed by the yeast cells and act as biological catalysts in the fermentation process are Maltase, Invertase and Zymass complex. Maltase has the ability to convert maltose, which is produced by starch degradation by alpha- and beta-amylases, to glucose and acts when the supply of simple sugars has been exhausted. Invertase converts sucrose to glucose and fructose, while the activity of the zymass complex results in the conversion of glucose, fructose and other simple sugars into carbon dioxide and ethanolAs described previously, it is carbon dioxide which causes the dough to rise.

Figure 3.1 CO2 produced by the yeast causes dough to rise Commercial yeast production starts in the laboratory where a small quantity of a yeast culture is injected into a closed flask containing a sterile solution of molasses, ammonium salts to provide a source of nitrogen, and phosphate, necessary for yeast development and reproduction. The yeast culture is made up of a particular yeast strain, which is normally kept on an agar slant. An enormous number of strains of Saccharomyces cerevisiae exist, many of which have already been selected for baking. The closed flask that the culture is injected into is kept at a constant temperature and the yeast grows vigorously for 12 hours. It is then transferred to a larger flask containing a further solution of molasses and nutrient material and more growth takes place. The transfer process is repeated again until a large enough culture of yeast is obtained to start the main yeast production process in the factorys large fermentation vessels. Fermentation vessels for yeast production range from 40,000 to 200,000L. The progressive increase of fermentor size used is known as scale-up. Until this stage the yeast cultures have been grown in the absence of air; this is known as anaerobic fermentation. Anaerobic fermentation is, however, inefficient in terms of yeast growth, and subsequent stages of yeast production take place with sterile air being blown/sparged through the growing yeast cultures; this is known as aerobic fermentation. The reason why the early stages of yeast production take place in the absence of air is to favour the growth of yeast cells instead of other organisms, such as bacteria, which may gain access to the culture, since these would also grow rapidly and could decrease the efficiency of the process and affect the final yeast quality. A small amount of alcohol is produced during the early stages, which inhibits the growth of foreign organisms.

The fermentation process continues with air being blown through the yeast cultures and molasses solution and nutrients being added continuously, at a constantly increasing rate that is directly proportional to the yeast cell population. By maintaining this supply level just sufficient for the amount of yeast present, together with an adequate supply of air, maximum yeast cell reproduction takes place with the minimum production of alcohol as indicated previously. At the end of the first stages of yeast growth about 12 tonnes of yeast is produced and this is known as seed or mother yeast. The seed yeast is divided into portions and these are used to start other fermentations. These fermentations are carried on as before with increasing addition of air, molasses solution and nutrients, and each 3 tonnes of seed yeast produces about 11 tonnes of the final bakers yeast. Throughout the whole fermentation process stringent checks are carried out to ensure that yeast growth and quality are maintained, so that the final 40-50 tonnes of bakers yeast are of the same quality and have the same characteristics and properties as the original few milligrams of pure yeast culture that started the process. At the end of the fermentation stage the yeast is present as a suspension of cells in a dark brown liquid containing the residues of the molasses. The yeast is removed from the fermentation liquid by a process of washing and separating in centrifugal separators, signaling the end of the fermentation and beginning of the downstream processing stage The fermentation of bakers yeast is strongly directed towards maximum biomass production, no byproducts such as ethanol are desired and so the fermentations are sectioned to obtain this max biomass.

Figure 3.3 The diagram of yeast fermentation

3.4 Yield
The whole processing will be yielded : Biomass 2 ethanol CO2 H2O

The bakers yeast production mainly desire biomass as the product, and however the additional production of ethanol is unwanted and the production of CO2 is needed to make the dough rise.

3.5 Downstream Processing

Downstream Processing can be defined as the stages of processing that take place after the fermentation or bioconversion stage.The yeast broth which is produced by fermentation, containing approximately 5% solids, can be manipulated into two main types of bakers yeast product and an additional intermediate saleable product. These are cake yeast, granular yeast and cream yeast, each of which requires a downstream process to arrive at the desired product. 3.5.1 Cream Yeast Cream yeast is not typically termed a bakers yeast product but is relevant as it represents a major step in the process and is a marketable product itself. At the end of the fermentation, the fermentor/yeast broth is concentrated using a series of combined centrifugation and washing steps, into a yeast cream with a solids concentration of approximately 20%. The yeast is then cooled to approximately 4C, an ideal temperature to restrict the growth of any contaminating mesophilic microorganisms. The cooled yeast cream is stored in a stainless steel cream tank, which is insulated and equipped with agitators and cooling pipes, effectively preventing heat exchange with the surrounding atmosphere, keeping the cream at 4C. Following storage either of two pathways can be followed. The first involves the preparation for sale of the cream yeast itself. Cream Yeast is basically the liquid product and can therefore be transferred into sterile tanks/containers and distributed to bakeries, where it is used to produce yeast based products. The advantage of this is that it excludes any human handling and therefore reduces the risk of contamination by handling, however due to its high (water) volume, transport costs can be expensive. For this reason, distribution is generally confined to a particular area.

3.5.2 Granular Yeast Granular Yeast, also known as Instant Dried Yeast, is a form of compressed yeast. Stored cream/liquid yeast is passed through a filter, usually a filter press or rotary vacuum filter, which removes water increasing its solids content to approximately 30%. Salt may also be added to the cream yeast prior to filtration to aid the removal of water. The filtered yeast is then dried using fluid-bed dryers. As the yeast is dry it generally does not require refrigeration as the low water content reduces the risk of microbial contamination. Emulsifiers and oils can be added at this point to texturize the yeast and aid the cutting process. As the name implies, granular yeast is crumbled into granules, the granulation process being carried out by a granulator. Granular broths are typically used to make restoring drinks to serve in a cup; the practicality of granular products coming both from their instantly soluble nature and the fact that they are easily measured. 3.5.3 Cake Yeast The filtered and dried yeast can alternatively be used to make cake yeast. Cake yeast is another form of compressed yeast and can be categorized as active dry yeast. It Page 9 of 12 differs from granular yeast in that rather than granulation, the dried yeast is extruded or cut into blocks/cakes. Similar to granular yeast cake yeast also contains about 30% solids (70% water). The composition of solids may vary depending on the growth rate of the yeast as lower growth rates give lower protein, lower activity, higher carbohydrate, and higher stability.12 Both types of compressed yeast are then packaged, typically vacuum packed to reduce the risk of contamination by aerobic bacteria, and distributed to wholesalers or traders. The shelf life of Active Dry/Cake Yeast and Instant Dry/Granular Yeast at ambient temperature is 1 to 2 years.

3.6 Idea of Improvement

The idea of improvement on this bacteria is to try to invent this bacteria that could produce CO2 more rapidly. Research that has been carried out by the Tokyo laboratory of Yeast and Fermentation showed that new strains of bakers yeast that produce CO2 more rapidly, are more resistant to stress or produce proteins or metabolies that can modify bread flavor, dough rheology or shelf-life. So, the development of baking industries could be wider and have more variety whether it is in ther flavor, texture or shelflife.

The other improvement is to recycle the waste of bread and product return from bakery industries. The waste of bread can be watering into a mash and trying to extract desirable components, for example like liquid sour, ethanol or carbon dioxide. But this action can only be undergo in sterile conditions.

3.7Comment on the production overview

After I knew the detail process about the production process of bakery industries using saccharomyces cerevisiae bacteria, I think this is very interesting because we could learn how really small organism could help and plays an important role in food industries. Additionally, it has many advantages not only in one food product, but also has function in several food products.

4. References
http://en.wikipedia.org/wiki/Saccharomyces_cerevisiae http://www.microbiologybytes.com/video/Scerevisiae.html http://www.dcu.ie/~oshead/BE401/lectures/pres4382018852bea.pdf http://primephysique.com/Portals/46981/images/alcohol%20fermentation%20process.gif http://www.desktopclass.com/education/9th-10th/genetic-engineering-and-biotechnology-lesson20-1.html