Effect Of Temperature On The Growth Of Yeast

Project and Seminar Material for Science Laboratory Technology SLT

Abstract

The effects of temperature on the growth of yeast and on its metabolic activity in distiller’s malt wort have been studied. In un-aerated fermentations, maximum yeast production takes place at about 30° C. whereas the growth rate in aerated cultures is highest at 35° C. The lag phase of the yeast studied fell from 6 hr. at 20° C. to 2-8 hr. at 25° C. and was not thereafter greatly affected by increases of temperature until 42° C. was reached, at which point growth ceased. Maltase activity was maximal at 25° C. when considered in terms of unit quantities of either yeast or fermenting wort, but the optimum temperature for initial fermentation velocity varied according to the time over which the measurement was made, being maximal at 40° C. for 0-5 hr., and at 35° C. for 2 hr. Alcohol production was highest at 25° C. whereas glycerol and higher alcohol formation took place optimally at 30° C.

Chapter One

1.0 Introduction

The word “Yeast” comes to us from the old English gist, gyst and from the indo-European root yes-meaning boil, from or bubble. Yeast microbes are probably one of the earliest domesticated organizing. People have used yeast for fermentation and baking throughout history. Archaeologists digging in Egyptian ruins found early grinding stones and baking chambers for yeasted bread, as well as drawings of 4,000. years- old bakeries and breweries. (Phillips 2012) in 1680, the Dutch naturalist Anton van leeuwenhock first microscopically observed yeast but at the time he did not consider them to be living organisms but rather globular structures (Huxley, 1871) in 1857 French microbiologist louis Pastear. Proved in the paper “memire sur la fermentation alcoolique” that alcoholic fermentation was conducted by living yeasts and not by a chemical catalyst. (Barntt 2003). Pasteur showed that by bubbling oxygen into the yeast broth, cell growth could be increased but fermentation was inherited as an observation cater called the “Pasteur effect”

1.1 Yeast

Yeast is any of various one called fungi that can cause the fermentation of carbohydrates producing carbon dioxide and alcohol. Bakers put it in bread to make it rise. Yeast is also used for the production of beer and wine. Yeast that are used commercially consist of masses of microscopic yeast organism.

1.1.1 Types of Yeast

Yeats is a very interesting little organism. It is neither a plant nor an animal but a single called fungus. As such there are several different species and varieties of yeast. And just like the apples you choose to make year cider, each one will make the end product taste a bit different. The difference in come is because of the rate of sugar consumption and carbon dioxide and other waste product creation as well as temperature and PH needed for fermentation.

The two main types of yeast are:

Baker’s yeast
Brewer’s yeast.

While both are saccharomyces cerevisiae, they are different strains (Like the difference between red delicious and Macintosh applies). The active dry yeast that you can get in grocery store is bakers yeast and is used for baking bread. To brew hard cider you want brewers yeast. This comes in dry and liquid forms.

1.1.2 Importance of Yeast

Yeast is important because it is used for making food which people eat. Yeast is used in the making of bread beer and wine. Bread in particular is an extremely important part of people diets world wide.

1.1.3 How Yeast is Used

Yeast lack chlorophyll. The lack of chlorophyll in yeast means that yeast has to rely on other things for food. Fruit grain, nectar and molasses are the things yeast feeds on for sugar. Yeast break down its food by producing a chemical called ferments or enzymes. Some species of yeast break down their food into alcohol and carbon dioxide.

That process is called fermentation

Fermentation is important in making bread, beer and wine.

Temperature: Temperature is the measurement in degrees of how hot or cold a thing or place is (Oxford dictionary 6th edition 2000).

1.1.4 Yeast Production

General

Baker’s yeast is currently manufactured in the United States at 13 plants owned by 6 major companies. Two main types of baker’s yeast are produced, compressed (cream) yeast and dry yeast. The total U. S. production of baker’s yeast in 1989 was 223,500 megagrams (Mg) (245,000 tons). Of the total production, approximately 85 percent of the yeast is compressed (cream) yeast, and the remaining 15 percent is dry yeast. Compressed yeast is sold mainly to wholesale bakeries, and dry yeast is sold mainly to consumers for home baking needs. Compressed and dry yeasts are produced in a similar manner, but dry yeasts are developed from a different yeast strain and are dried after processing. Two types of dry yeast are produced, active dry yeast (ADY) and instant dry yeast (IDY). Instant dry yeast is produced from a faster-reacting yeast strain than that used for ADY. The main difference between ADY and IDY is that ADY has to be dissolved in warm water before usage, but IDY does not.

1.1.5 Process Description

The first stage of yeast production consists of growing the yeast from the pure yeast culture in a series of fermentation vessels. The yeast is recovered from the final fermentor by using centrifugal action to concentrate the yeast solids. The yeast solids are subsequently filtered by a filter press or a rotary vacuum filter to concentrate the yeast further. Next, the yeast filter cake is blended in mixers with small amounts of water, emulsifiers, and cutting oils. After this, the mixed press cake is extruded and cut. The yeast cakes are then either wrapped for shipment or dried to form dry yeast.

1.1.6 Raw Materials

The principal raw materials used in producing baker’s yeast are the pure yeast culture and molasses. The yeast strain used in producing compressed yeast is Saccharomyces cerevisiae. Other yeast strains are required to produce each of the 2 dry yeast products, ADY and IDY. Cane molasses and beet molasses are the principal carbon sources to promote yeast growth. Molasses contains 45 to 55 weight percent fermentable sugars, in the forms of sucrose, glucose, and fructose. The amount and type of cane and beet molasses used depend on the availability of the molasses types, costs, and the presence of inhibitors and toxins. Usually, a blend consisting of both cane and beet molasses is used in the fermentations. Once the molasses mixture is blended, the pH is adjusted to between 4.5 and 5.0 because an alkaline mixture promotes bacteria growth. Bacteria growth occurs under the same conditions as yeast growth, making pH monitoring very important. The molasses mixture is clarified to remove any sludge and is then sterilized with high-pressure steam. After sterilization, it is diluted with water and held in holding tanks until it is needed for the fermentation process. A variety of essential nutrients and vitamins is also required in yeast production. The nutrient and mineral requirements include nitrogen, potassium, phosphate, magnesium, and calcium, with traces of iron, zinc, copper, manganese, and molybdenum. Normally, nitrogen is supplied by adding ammonium salts, aqueous ammonia, or anhydrous ammonia to the feedstock. Phosphates and magnesium are added, in the form of phosphoric acid or phosphate salts and magnesium salts. Vitamins are also required for yeast growth (biotin, inositol, pantothenic acid, and thiamine).

Thiamine is added to the feedstock. Most other vitamins and nutrients are already present in sufficient amounts in the molasses malt.

1.1.7 Fermentation

Yeast cells are grown in a series of fermentation vessels. Yeast fermentation vessels are operated under aerobic conditions (free oxygen or excess air present) because under anaerobic conditions (limited or no oxygen) the fermentable sugars are consumed in the formation of ethanol and carbon dioxide, which results in low yeast yields.

The initial stage of yeast growth takes place in the laboratory. A portion of the pure yeast culture is mixed with molasses malt in a sterilized flask, and the yeast is allowed to grow for

2 to 4 days. The entire contents of this flask are used to inoculate the first fermentor in the pure culture stage. Pure culture fermentations are batch fermentations, where the yeast is allowed to grow for 13 to 24 hours. Typically, 1 to 2 fermentors are used in this stage of the process. The pure culture fermentations are basically a continuation of the flask fermentation, except that they have provisions for sterile aeration and aseptic transfer to the next stage.

Following the pure culture fermentations, the yeast mixture is transferred to an intermediate fermentor that is either batch or fed-batch. The next fermentation stage is a stock fermentation. The contents from the intermediate fermentor are pumped into the stock fermentor, which is equipped for incremental feeding with good aeration. This stage is called stock fermentation, because after fermentation is complete, the yeast is separated from the bulk of the fermentor liquid by centrifuging, which produces a stock, or pitch, of yeast for the next stage. The next stage, pitch fermentation, also produces a stock, or pitch, of yeast. Aeration is vigorous, and molasses and other nutrients are fed incrementally. The liquor from this fermentor is usually divided into several parts for pitching the final trade fermentations (adding the yeast to start fermentation). Alternately, the yeast may be separated by centrifuging and stored for several days before its use in the final trade fermentations. The final trade fermentation has the highest degree of aeration, and molasses and other nutrients are fed incrementally. Large air supplies are required during the final trade fermentations, so these vessels are often started in a staggered fashion to reduce the size of the air compressors. The duration of the final fermentation stages ranges from 11 to 15 hours. After all of the required molasses has been fed into the fermentor, the liquid is aerated for an additional 0.5 to 1.5 hours to permit further maturing of the yeast, making it more stable for refrigerated storage.

The amount of yeast growth in the main fermentation stages described above increases with each stage. Yeast growth is typically 120 kilograms (270 pounds) in the intermediate fermentor, 420 kilograms (930 pounds) in the stock fermentor, 2,500 kilograms (5,500 pounds) in the pitch fermentor, and 15,000 to 100,000 kilograms (33,000 to 220,000 pounds) in the trade fermentor. The sequence of the main fermentation stages varies among manufacturers. About half of existing yeast operations are 2-stage processes, and the remaining are 4-stage processes. When the 2-stage final fermentation series is used, the only fermentations following the pure culture stage are the stock and trade fermentations. When the 4-stage fermentation series is used, the pure culture stage is followed by intermediate, stock, pitch, and trade fermentations.

1.1.8 Harvesting and Packaging

Once an optimum quantity of yeast has been grown, the yeast cells are recovered from the final trade fermentor by centrifugal yeast separators. The centrifuged yeast solids are further concentrated by a filter press or rotary vacuum filter. A filter press forms a filter cake containing 27 to 32 percent solids. A rotary vacuum filter forms cakes containing approximately 33 percent solids. This filter cake is then blended in mixers with small amounts of water, emulsifiers, and cutting oils to form the end product. The final packaging steps, as described below, vary depending on the type of yeast product. In compressed yeast production (SCC 3-02-035-XX), emulsifiers are added to give the yeast a white, creamy appearance and to inhibit water spotting of the yeast cakes. A small amount of oil, usually soybean or cottonseed oil, is added to help extrude the yeast through nozzles to form continuous ribbons of yeast cake. The ribbons are cut, and the yeast cakes are wrapped and cooled to below 8°C (46°F), at which time they are ready for shipment in refrigerated trucks.

In dry yeast production (SCC 3-02-034-XX), the product is sent to an extruder after filtration, where emulsifiers and oils (different from those used for compressed yeast) are added to texturize the yeast and to aid in extruding it. After the yeast is extruded in thin ribbons, it is cut and dried in either a batch or a continuous drying system. Following drying, the yeast is vacuum packed or packed under nitrogen gas before heat sealing. The shelf life of ADY and IDY at ambient temperature is 1 to 2 years.

Volatile organic compound (VOC) emissions are generated as byproducts of the fermentation process. The 2 major VOCs emitted are ethanol and acetaldehyde. Other byproducts consist of other alcohols, such as butanol, isopropyl alcohol, 2,3-butanediol, organic acids, and acetates. Based on emission test data, approximately 80 to 90 percent of total VOC emissions is ethanol, and the remaining 10 to 20 percent consists of other alcohols and acetaldehyde. Acetaldehyde is a hazardous air pollutant as defined under Section 112 of the Clean Air Act. Volatile byproducts form as a result of either excess sugar (molasses) present in the fermentor or an insufficient oxygen supply to it. Under these conditions, anaerobic fermentation occurs, breaking down the excess sugar into alcohols and carbon dioxide. When anaerobic fermentation occurs, 2 moles of ethanol and 2 moles of carbon dioxide are formed from 1 mole of glucose. Under anaerobic conditions, the ethanol yield is increased, and yeast yields are decreased. Therefore, in producing baker’s yeast, it is essential to suppress ethanol formation in the final fermentation stages by incremental feeding of the molasses mixture with sufficient oxygen to the fermentor.

The rate of ethanol formation is higher in the earlier stages (pure culture stages) than in the final stages of the fermentation process. The earlier fermentation stages are batch fermentors, where excess sugars are present and less aeration is used during the fermentation process. These fermentations are not controlled to the degree that the final fermentations are controlled because the majority of yeast growth occurs in the final fermentation stages. Therefore, there is no economical reason for manufacturers to equip the earlier fermentation stages with process control equipment. Another potential emission source at yeast manufacturing facilities is the system used to treat process waste waters. If the facility does not use an anaerobic biological treatment system, significant quantities of VOCs could be emitted from this stage of the process. For more information on waste water treatment systems as an emission source of VOCs, please refer to EPA’s Control Technology Center document on industrial waste water treatment systems, Industrial Wastewater Volatile Organic Compound Emissions – Background Information For BACT/LAER, or see Section 4.3 of AP-42. At facilities manufacturing dry yeast, VOCs may also be emitted from the yeast dryers, but no information is available on the relative quantity of VOC emissions from this source.

1.1.9 Controls
Only 1 yeast manufacturing facility uses an add-on pollution control system to reduce VOC emissions from the fermentation process. However, all yeast manufacturers suppress ethanol formation through varying degrees of process control, such as incrementally feeding the molasses mixture to the fermentors so that excess sugars are not present, or supplying sufficient oxygen to the fermentors to optimize the dissolved oxygen content of the liquid in the fermentor. The adequacy of oxygen distribution depends upon the proper design and operation of the aeration and mechanical agitation systems of the fermentor. The distribution of oxygen by the air sparger system to the malt mixture is critical. If oxygen is not being transferred uniformly throughout the malt, then ethanol will be

produced in the oxygen-deficient areas of the fermentor. The type and position of baffles and/or a highly effective mechanical agitation system can ensure proper distribution of oxygen. A more sophisticated form of process control involves using a continuous monitoring system and feedback control. In such a system, process parameters are monitored, and the information is sent to a computer. The computer is then used to calculate sugar consumption rates through material balance techniques. Based on the calculated data, the computer continuously controls the addition of molasses. This type of system is feasible, but it is difficult to design and implement. Such enhanced process control measures can suppress ethanol formation from 75 to 95 percent. The 1 facility with add-on control uses a wet scrubber followed by a biological filter. Performance data from this unit suggest an emission control efficiency of better than 90 percent.

1.10 Emission Factor

Table 9.13.4-1 provides emission factors for a typical yeast fermentation process with a moderate degree of process control. The process emission factors in Table 9.13.4-1 were developed from 4 test reports from 3 yeast manufacturing facilities. Separate emission factors are given for intermediate, stock/pitch, and trade fermentations. The emission factors in Table 9.13.4-1 are expressed in units of VOC emitted per fermentor per unit of yeast produced in that fermentor. In order to use the emission factors for each fermentor, the amount of yeast produced in each fermentor must be known. The following is an example calculation for a typical facility:

			Total Yeast
			Production Per
		No. Of Batches	Stage, tons/yr
Fermentation	Yeast Yield Per	Processed Per	(C = A x	Emission Factor,	Emissions, lb	Percent of Total
Stage	Batch, lb (A)	Year, #/yr (B)	B/2,000)	lb/ton (D)	(E = C x D)	Emissions

Intermediate	265	156	21	36	756	0.84
Stock	930	208	97	5	485	0.54
Pitch	5,510	208	573	5	2,865	3.18
Trade	33,070	1,040	17,196	5	85,980	95.44
TOTAL	Ð	Ð	Ð	Ð	90,086	100

In most cases, the annual yeast production per stage will not be available. However, a reasonable estimate can be determined based on the emission factor for the trade fermentor and the total yeast production for the facility. Trade fermentors produce the majority of all VOCs emitted from the facility because of the number of batches processed per year and of the amount of yeast grown in these fermentors. Based on emission test data and process data regarding the number of batches processed per year, 80 to 90 percent of VOCs emitted from fermentation operations are a result of the trade fermentors. Using either a 2-stage or 4-stage fermentation process has no significant effect on the overall emissions for the facility. Facilities that use the 2-stage process may have larger fermentors or may produce more batches per year than facilities that use a 4-stage process.The main factors affecting emissions are the total yeast production for a facility and the degree of process control used.

Table 2 (Metric And English Units). VOLATILE ORGANIC COMPOUND (VOC) EMISSION FACTORS FOR YEAST MANUFACTURING

EMISSION FACTOR RATING:

		VOC^c
	VOC Emitted Per Stage Per		VOC Emitted Per Stage Per
	Amount Of Yeast Produced		Amount Of Yeast Produced
Emission Point^b	In A Stage,		In A Stage,
Emission Point^b	kg VOC/Mg Yeast		lb VOC/ton Yeast
Fermentation stages^d
Flask (F1)	ND		ND
Pure culture (F2/F3)	ND		ND
Intermediate (F4)	18		36
(SCC 3-02-034-04)
Stock (F5)	2.5		5.0
(SCC 3-02-034-05)
Pitch (F6)	2.5		5.0
(SCC 3-02-034-06)
Trade (F7)	2.5		5.0
(SCC 3-02-034-07)
Waste treatment
(SCC 3-02-034-10)	See Section 4.3 of AP-42
Drying
Drying
(SCC 3-02-034-20)	ND		ND

^a References 1,6-10.
Total VOC as ethanol
SCC = Source Classification Code.
ND = no data.

F numbers refer to fermentation stages

Factors are for both dry yeast (SCC 3-02-034-XX) and compressed yeast (SCC 3-02-035-XX).
Factors should be used only when plant-specific emission data are not available because of the high degree of emissions variability among facilities and among batches within a facility.
Some yeast manufacturing facilities use a 2-stage final fermentation process, and others use a 4-stage final fermentation process. Factors for each stage cannot be summed to determine an overall emission factor for a facility, since they are based on yeast yields in each fermentor rather than total yeast production. Total yeast production for a facility equals only the yeast yield from the trade fermentations. Note that CO2 is also a byproduct of fermentation, but no data are available on the amount emitted.

Chapter Five

5.0 Conclusion and Summary

Conclusion and Summary

Yeast is useful and important to humans in many ways. Temperature is very important to yeast growth. Yeast is used in bread, which is imperative because most people eat bread. We could live without bread but it is still important worldwide.

Conclusions

S. cerevisiae(yeast) has long been utilized as a functional food and supplement. A number of biologically active secondary metabolites including naringenin, reticuline, artemisinin, PAI, and other pigments have been isolated from S. cerevisiae. S. cerevisiae produces pigments and other bioactive secondary metabolites that have been shown to exhibit neuroprotective, antioxidant, antidiabetic, antiinflammatory, antimalarial, and antitumoral properties. S. cerevisiae has potential safety concerns due to its levels of aflatoxin which has nephrotoxic, hepatoxic, and cytotoxic effects. Therefore, S. cerevisiae is a potential bioactive agent and food supplement that requires further clinical studies and development