___BE401____

The Production and Downstream Processing of Bakers Yeast

Authors P. Weston E. Mooney Module Degree Programme Year BE401 B.Sc. in Biotechnology 4 52393654 52058294

The aim of this paper was to investigate the production and downstream processing of bakers yeast. Bakers yeast is a worldwide commercial product. It is used in bread making to leaven and aerate dough. There is a large market for baked goods with a large profit to be made. Bakers yeast is made up of the living cells of the microscopic unicellular fungus saccharomyces cerevisiae. The fermentations focus on a maximum biomass yield with limited ethanol production, favouring the oxidative or aerobic metabolic yeast pathway. 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. This cream can be sold as cream yeast or further filtered, de-watered, extruded and packaged for sale as high activity dried yeast or cake yeast (Att. 3). The process, as with all biotechnology processes, is constantly undergoing research and development to optimize the process technology and its components, ultimately producing faster growing strains and better quality end products.

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Introduction
Bakers Yeast is a commercial preparation consisting of dried cells of one or more

strains of the fungus Saccharomyces cerevisiae. Bakers use yeast as a leavening agent in the rising of dough for baking. A secondary contribution of yeast to bread is flavouring and aroma. Bakers yeast is a high volume, low value product, with 1574 x 106 kg being produced per annum on a global scale.1 Bakers Yeast is marketed in two ways, either as compressed cakes or as a dry powder, however there is also a saleable intermediate of the process known as cream yeast. These will be discussed later. Process considerations include media formulation (which has to be cost effective), and the limited respiratory capacity of yeast, which inhibits the production of biomass in favour of ethanol production. 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 1.1 below shows how the series of fermentations carried out progress from batch to fed batch in order to maximize biomass yield.

Figure 1.1 The fermentative capacity of bakers yeast is strongly dependent on the final specific growth rate during biomass production in high-cell-density fed-batch cultures.2 Bakers yeast today is a product of biochemical, microbiological, technical knowledge and experience. Biochemistry has led to an insight into the fermentation process; Page 2 of 12

microbiology has made it possible to breed new and better strains of yeast and to develop better techniques for sterilization and disinfection. Advancing technologies have led to the large scale production of yeast with a high degree of automation and process control, giving commercial yeast of consistent quality and activity at an economic price (Att.1). By feeding on sugars from the starch in flour, yeast produces carbon dioxide. This gas expands the gluten proteins in the flour and causes the dough to rise, this process of bread making being the most commonly associated with yeast. Scientists now cultivate strains of bakers yeast for their ability to make dough rise and produce loaves of good height, texture and flavour.3

Saccharomyces cerevisiae
Yeast are a unicellular fungi or plant-like microorganism that exists in or on all living

matter i.e. water, soil, plants, air, etc. They are a microbial eukaryote, associated with ascomycetes and are rich in protein and vitamin B. 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. There are hundreds of different species of yeast identified in nature, but the genus and species most commonly used for baking is Saccharomyces cerevisiae. The scientific name Saccharomyces cerevisiae, means 'a mold which ferments the sugar in cereal (saccharo-mucus cerevisiae) to produce alcohol and carbon dioxide.4 Yeasts are usually spherical, oval or cylindrical in shape and a single cell of S. cerevisiae is around 8m in diameter. Each cell has a double-layered wall, which is permeable to certain substances and in this way food material is taken into the cell and metabolites leave it (see Fig.2.1 below). Cell division or cell reproduction generally takes place by budding. In the budding process, a new cell forms as a small outgrowth of the old cell, the bud gradually enlarges and then separates. Although most yeasts reproduce only as single cells, under some conditions some yeasts can form filaments.5 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.5

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Figure 2.1 Diagram of a typical yeast cell.4 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 ethanol (Att.1). As described previously, it is carbon dioxide which causes the dough to rise.

Biochemistry of Bakers Yeast Growth

S. Cerevisiae primarily uses molasses as its growth substrate (discussed later), 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

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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. Figure 3.1 illustrates the possible pathways in the growth of S. Cerevisiae to produce biomass (C1HHXOOXNNX). Subscripts HX, OX and NX vary according to the elemental composition of the biomass, which is dictated by such parameters as the rate of glucose/oxygen uptake and the production of ethanol which can have inhibitory effects. Sucrose (C12H22O11) Glucose/Fructose (C6H12O6) O2 NH3 CO2 H2O Biomass (C1HHXOOXNNX) Oxido-Reductive pathway Oxidative pathway Reductive pathway NH3 CO2 H2O Biomass (C1HHXOOXNNX) & Ethanol (C2H6O) O2 NH3 CO2 H2O Biomass (C1HHXOOXNNX) Figure 3.1 Simplified growth pathways of Saccharomyces cerevisiae.6

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Production of Yeast Fermentation & Scale-up

The basic stages of bakers yeast production are outlined in this section. Molasses is the

substrate used in the production of bakers 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. 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 baking19. 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 at Page 6 of 12

a 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.

Downstream Processing (DSP)

Downstream Processing can be defined as the stages of processing that take place after

the fermentation or bioconversion stage.7 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. These downstream processes are investigated in the subsections below.

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%.8 The yeast is then cooled to approximately 4C, an ideal

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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 pipes9, 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.10 The other pathway further manipulates the yeast cream into compressed or dried yeast as described in sections 5.2 and 5.3, these being the two forms of bakers yeast.

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.11

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

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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.

Cost and Profitability

Automation reduces the risk of contamination and human error and in particular the

need for a large workforce. It reduces labour costs for internal transporting and handling, concentrating its primary labour to system operators and maintenance. Automation also leads to the most efficient use of raw materials as automated systems can be calibrated to ensure that the optimum feed rate is delivered, eliminating any over or under-usage which human error may incur. Cream yeast systems range in cost from less than $300,000 (250,000) to more than $600,000 (500,000), depending on tank size, number of dosing stations, and amount of site preparation. The annual operating costs of the fermentation processes were estimated at around $111,000 (93,000) per year, (1992).13 There is a large market for baked goods and profits can range from 100-300 million for companies such as Delifrance or United States Bakery who produce baked goods among many others or 20-50 million for companies such as Panera Bread Company which focuses more in fresh baked goods. The market for bread and bakery products has recently recovered from a period of relative decline during the late 1990s, brought about by price cutting by retailers. During 2002, the market grew by 2%, reaching a total of 2.95bn (4.3bn).14 Many international and Irish companies are involved in the production of yeast based products, one of which is Proofex.

Irish Based Firm

Proofex was established in 1985 and operates from a state of the art high technology

factory located at Finglas, Dublin, which was custom built for the purpose of its

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operations. It operates under a number of formal quality management standards, such as ISO 9002 (a standard relating to the effectiveness of meeting customer needs) and was also awarded The Excellence through People Standard. It produces fresh bakers yeast supplying its products to Bakers, Pizza producers and Distillers in Ireland. Proofex supplies both compressed and fresh cream yeast, compressed being the preferred choice for smaller users. Proofex makes a point of informing its customers that the Proofex yeast strain, Saccharomyces cerevisiae, has not been produced by any genetic modification/ engineering techniques or recombinant DNA technology, submitting to the fears of consumers who are disinclined towards GM-associated products.

Current Trends and the Future of the Industry

Research has been conducted into recycling bakery product returns and waste.15 Bread

is commonly returned to industrial bakeries when it is unsold and used instead as animal feed. The research aimed to investigate the reuse of the bread by watering it into a mash and trying to extract desirable components e.g. bakers yeast, liquid sour, ethanol or carbon dioxide. It was however demonstrated that the installation cannot be operated under largely unsterile conditions as had originally been intended, unsterile conditions being of great importance as the commercial yeast industry cannot afford the sterile environment used in the pharmaceutical and enzyme industries.16 Research is constantly being carried out to further automate the process17 and genetically engineer the yeast cell.18 Recombinant-DNA technology has led to dramatic changes in formulation, ingredients or processing conditions. New strains of bakers yeast that produce CO2 more rapidly, are more resistant to stress or produce proteins or metabolites that can modify bread flavour, dough rheology or shelf-life are now emerging18 such as is being carried out by the Tokyo Laboratory of Yeast and Fermentation.

Conclusions
The bakers yeast industry is a major market, grossing several billion euros per year.

The low value, high volume product is produced under stringent environmental conditions to obtain the maximum product/biomass yield which is dependent on process

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design, cost and the strain of S. cerevisiae used. The fermentation and scale-up stage is the critical to the process as it dictates the volume and quality of the product. The industry is expanding after a recent slump and with the potential for large profits it can be expected to continue. As with all biotechnology processes, research and development in this area is continuously ongoing to create more advantageous/beneficial strains of the S. cerevisiae fungi and optimize the fermentation and processing steps which make up the bakers yeast process.

10 Appendices
10.1 Bibliography
1.

OShea, D., 2005, BE401 Industrial Bioprocessing Module Notes [online] Available from: www.dcu.ie/~oshead/BE401 [accessed 4 Oct 05]

2.

Van Hoek P., Aristidou A., Hahn J., Patist A., 2003. Fermentation Goes Large-Scale [online]. Available from: http://bio.aiche.org/pdfs/Articles/FERMEN~1.PDF. [accessed 04Oct 2005].

3.

The European Food Information Council (Eufic), 2005, Yeast The Illustrious Microbe [online]. Available form: http://www.eufic.org/gb/food/pag/food22/food222.htm [accessed 4 Oct 2005]

4.

The Artisan, The Yeast Treatise, 2002, Yeast Production - General Discussion [online]. Available from: http://www.theartisan.net/yeast_treatise_frameset.htm [accessed 4 Oct 2005] Madigan M.T., Martinko J.M. and Parker J. (2003) Brock Biology of Microorganisms, 10th Edition, Pearson Education Inc.

5.

6.

Sonnleitner B, Kappeli O., 1986. Gowth Of Saccharomyces-Cerevisiae Is Controlled By Its Limited Respiratory Capacity - Formulation And Verification Of A Hypothesis. Biotech and Bioeng, 28 (6): 927-937 Jun 1986.

7.

Definitions of Downstream processing on the Web [online], 2005. Available from: www.siebelinstitute.com/brewterms/D.html [accessed 07Oct 2005]

8.

Team Minn-Dak Yeast, 2005. Bakers Yeast Production [online]. Available from: http://www.dakotayeast.com/yeast_production.html. [accessed 17Oct 2005].

9.

Kristiansen, B., 1994. Integrated design of a fermentation plant : the production of baker's yeast. Weinheim, N.Y.

10.

Lallemand Inc., 2001. Cream Yeast, Lallemand Baking Update, Volume 1/Number 2 [online]. Available from: http://www.lallemand.com/BakerYeastNA/eng/PDFs/LBU%20PDF%20FILES/1_2CREAM.PDF [accessed 24Oct 2005].

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11.

Bauer S.p.a, 2005. Bauer Products, Useful Information [online]. Available from: http://www.bauerspa.it/eng/prodotti/utili.htm [accessed 17Oct 2005].

12.

Lallemand Inc., 2001. Yeast Production, Lallemand Baking Update, Volume 1/Number 9 [online]. Available from: http://www.lallemand.com/BakerYeastNA/eng/PDFs/LBU%20PDF%20FILES/1_9YPROD.PDF [accessed 07Oct 2005].

13.

Assessment of VOC Emissions and Their Control from Bakers Yeast Manufacturing Facilities. Office of Air Quality Planning and Standards, U.S. Environmental Protection Agency, Research Triangle Park NC. EPA-450/3-91-027 January 1992.

14.

Research and Markets, 2003. Bread and Bakery Products Market Report [online]. Available from: http://www.researchandmarkets.com/reports/34771 [accessed 24Oct 2005].

15.

Dr. Ing. Reimelt S., Schraps S., 2001. Biotechnological Processing Of Bakery Product Returns And Waste [online]. Available from: http://asbe.org/program00/environment/reimeltfinal.html [accessed 17Oct 2005].

16.

Rehm H.J., Reed G., Puhler A and Stadler P. (1995) Biotechnology Enzymes, Biomass, Food and Feed, 2nd Edition, Vol. 9

17.

Ringbom K., Rothberg A., Saxn B. (1996). Model-based automation of bakers yeast production, J. Biotech. 51, 73-82.

18.

Randez-Gil F., Sanz P., Prieto J. A., (1999). Engineering bakers yeast: room for improvement, Tibtech. 17, 237-243.

19.

The Artisan, 2002. Yeast Production - General Discussion [online]. Available from: http://home.earthlink.net/~ggda/yeast_production_general_discussion.htm [accessed 2 Nov 05]

10.2 List of Attachments

Attachment 1 2 3 Description An Introduction to Bakers Yeast, Proofex. Yeast Products The Manufacture of Bakers Yeast, Proofex Yeast, Proofex

11 Acknowledgements
We would like to thank Proofex Yeast Products Company for their help and the information they supplied to us. Also we would like to thank Dr. Donal OShea for his guidance.

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