brewer yeast

1. 1
BORKHUU NOMIN-ERDENE
MAKING PRODUCTS USING FOOD WASTE
Supervisor’s: /Du Ming/
/D.Bolormaa/
2020

2. 2
Content
Introduction
Basis Research ........................ . ................. ............... .. ... …....... ... . ........... 4
Research goals and objectives ........ ... ...................... .................. .... ............. 5
Chapter One
1. Part of theory
1.1 Brewer's yeast ................................ .............. .......... ... ... ..... ………………..6
1.2 Chemical composition of yeast cell ............................................……….….....8
1.3 Yeast taxonomy .............................................. ... ... ... ... ...... . ……………….8
1.4 Conditions for yeast multiplication, growth and culture ........ ................. .... ..10
1.5 Studied………………………………………………………………………………16
Chapter Two
2. Research materials and methodology .................................................... .25
2.1 Microbiological analysis ............................................... ........ .......... ..............26
2.2 Moisture by drying (MNS 245-5:87)….............................................. ............. 27
2.3 Lowry’s method for protein determination .......................... ..... ... .. .... ……..27
2.4 Keldel method for protein determination (MNS 3746:84)................................27
2.5 Determination of carbohydrate volume...........................................................28
2.6 Method for determination of fat…………………………………........................29
2.7 Bertrand’s method for determination of sugar…………………………………..29
Chapter three
3. Discussion of research results……………………………..…………………30
3.1 Yeast cell growth density ................................................................................30
3.2 Determination of some chemical components of brewery waste ................... 31
3.3 The amount of yeast cell growth density ......................................................... 32
3.4 Research on the production of protein feed using brewery waste ...................33

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3.5 Some chemical parameters for culturing yeast strains using slag ......................35
3.6 Microbiological analysis ............................................... .. ....... ..........................37
Conclusion: ................................................ ............................................................ 38
References................ …................. ..................... ....................................................39
Appendix .................................................................................................................. 41

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INTRODUCTION
The rapid growth of the world's population is a challenge for every country to find new
sources of food and consequently, to provide humanity with safe food rich in protein, amino acids and
vitamins.
Nowadays, there is a need to run production without waste technology. Wastes from beer production
are slag and residues from production after brewing. These deals just to re-develop to explore
opportunities for e.g. a variety of products have become a pressing issue.
According to researchers, the volume of agricultural production is growing, but lags behind the
food needs of the population. Microbiological production is considered to play an important role in
improving food supply, which is one of the major social problems. First of all, it will have a significant
impact on many issues, such as protein deficiency, supply of vitamins, minerals and biologically active
substances. It is possible to produce them using a variety of microorganisms. At the same time, there is
a wide range of research on the development and implementation of non-waste technologies based on
the use of modern microorganisms , and the production of a wide range of microbiological food
carbohydrates, polysaccharides and medical products.
The production of proteins in microbial cells is very fast, for example, 10-100 thousand times
faster than in animals. Here is a classic example. A cow weighing 400 kg produces 400 grams of
protein per day, while the same amount of bacteria can " produce " 40,000 tons of protein in that time .
Industrial technology with microbial synthesis is less labor-intensive than agriculture and is not
affected by natural factors (heat and cold, drought, lighting, sunlight, etc) .
An international standard for the production of microbial food was developed in 1983. The US
Department of Food and Drug Administration, the Center for Nutrition, Food Safety, and the Food
Safety Authority have joined forces to create a list of safe strains of microorganisms to be used
worldwide in the manufacture of biologically active preparations and other products . released in
September (FDA, GRAS, CFR-21) . The list includes many species of bacteria, fungi and
yeast email is effective, aktinomitsyet identified including biologicals production .
Yeast strains include Saccharomyces cerevisiae, Sacc.fragilis, Sacc.lactis, Candida utilis,
C.guilliermondii, C.lipotytica, C. pseudotropicalis, Kluyveromyces marxianusvar.lactis., And
Kl.lactis .

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RATIONALE FOR THE RESEARCH
In recent years, the light food industry has been developing rapidly in our country, and due to
the lack of special ways to use the waste from the design of the first factory, the amount of waste is
increasing and polluting the environment. However, in most cases, food waste is the main raw material
for the production of high quality protein feed.
It is estimated that 29 breweries in Mongolia today produce about 4 million liters of beer a
year. The APU brewery alone emits 8,000 tons of beer slag and 375 tons of waste per year. The
production of various additives by microbiological processing of beer waste, which cannot be used
directly as animal feed, is not only important to reduce the negative impact on the environment, but
also to create a waste-free production technology.
There has been some research on the use of brewery waste in Mongolia, but almost no work
has been done on it. Beer slag is widely used in animal feed, biofertilizers, energy production,
microbiological research, mushroom growing medium, bricks, flour and bakery products . Because
beer slag contains biologically active substances, it is possible to produce the perfect food
biopreparation. For this reason , there is an urgent need to study the possibility of producing a variety
of products using waste from the brewery, and to develop technology for the production of food
additives ( biopreparations ) . Based on all of the above, we decided to conduct a study on the use
of waste ( beer slag ) from the brewery .
The purpose of the research
The purpose of the research is to study the possibility of making products using the waste of APU
Brewery in Ulaanbaatar. In order to achieve this goal, the following objectives have been set. These
include:
Objectives of the research
1. Determination of chemical composition (protein, carbohydrates, fats, ash) of APU LLC brewery
waste
2. Experiment with animal feed by culturing two strains of yeast using waste slag
3. Determination of some chemical components (proteins, carbohydrates, fats, ash) and
microbiological (contamination, cell count) of biopreparations derived from brewery waste.
4. Develop a technology scheme for protein feed

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CHAPTER ONE.
Theoretical part
1.1 Brewer's yeast
Yeast cell structure: Yeast does not form micelles and is immobile. A single-celled eukaryotic
microorganism without chlorophyll granules and is widespread. Many types of yeast are widely used
in the food industry and medicine. Yeast has the ability to break down sugars into ethyl alcohol and
carbon dioxide, which is why they are called mushrooms or saccharomycetes. Yeast is a unique type
of microorganism capable of producing large amounts of biologically active substances such
as proteins and vitamins ( B1, B2, B3, nicotinic acid ). Electron microscopy of yeast cells ( Figure 1 )
Figure1. Yeast cells
Yeast cells are usually round or slightly oval in shape, and occasionally cylindrical, sickle,
arrow, and triangular cells are found.
It is 10-15 microns in diameter. Yeast cell shape and size may vary depending on factors such
as cell age, appearance, and nutrition. As the cell ages, its size increases. The macromolecules that
make up yeast cells are proteins, glycoproteins, polysaccharides, polyphosphates, lipids, and nucleic
acids. (Table 1) Yeast cells are made up of two main components: protoplasts and membranes. The
cell membrane consists of several layers containing compounds such as polysaccharides and lipids.
Yeast protoplasts contain cytoplasmic membranes, ribosomes, mitochondria, and nuclei, as well as
pellets in the form of pellets, which contain various nutrient reserves, oil droplets, glycogen, and
pigments. Some types of yeast form mucous membranes that stick together to form a sponge-like
structure in a liquid medium and settle to the bottom of the container. They are called cotton yeasts,
and those that do not form a mucous membrane and spread evenly in a liquid medium are called
particulate yeasts.

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Macromolecular composition of yeast cells Table 1
Macromolecule Duties The main components
Protein Structural unit
Hormone
Enzyme
Actin, tubulin ( cytoskeleton ) , histone
( H2A, H2B, H3, H4 ) , ribosomal protein
to a pheromone
Glycoprotein Participate in cell wall
structure
Enzyme
Mann protein
The main enzyme of action
Inverters
Polysaccharide Participate in cell wall
structure
Reserves involved in the
structure of the capsule
Gluten, semolina, chitin
Glycogen, Trego lose
Polyphosphate Reserves Polyphosphate in the vacuole
Lipid Structural unit
Reserves
In action
Participate
Membrane-free sterols
Presence of lipids ( sterol esters and
triglycerides )
Phosphoglyceride derivatives, free fatty acids
Nucleic acid DNA
RNA
Genome DNA (80%)
Bamitochondrial DNA (10-20%)
Primary (80%) , Primary (5%) , Primary and
other forms of RNA

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1.2 Chemical composition of yeast cells
190-252 kg of protein biomass can be obtained by growing yeast using one ton of sawdust, straw and
plant stems. Capital proteins are no less biologically valuable than animal proteins, and contain 50-
60 % protein, of which 40-50 % is absorbed. The cost of 1 ton of feed yeast is cheaper than 1 ton of
liquid milk, which is rich in essential amino acids and a valuable source of nutrients.
Yeast biomass is similar in amino acid content to animal protein. 1 kg of feed yeast contains 35-
42 g of lysine, which is 10 times more than barley and oats. And methionine is 1.5-2.0 times
higher. The amount of tryptophan is 2-3 times higher than that of yeast biomass grains. However,
yeast protein is a source of these amino acids, as grasses are deficient in lysine and methionine
tryptophan. Candida yeast cells contain 20-35 % of carbohydrates and their absorption reaches 85-95 %
( sometimes 100 %) . 12-15 g of phosphorus, 3-8 g of calcium, 8-17 mg of iron, 16-30 mg of copper,
20-40 mg of manganese, 20-70 mg of iron, 1.4-2.2 mg of cobalt per 1 kg of yeast are easily absorbed
minerals. are included. Feed yeast proteins are rich in albumin, globulin, phosphoproteins,
nucleoproteins, lipoproteins, glycoproteins, soluble protein peptones, polypeptides, and amino
acids. Valine, leucine, isoleucine, and arginine, which are essential amino acids in feed yeast, are
considered to be more like beer than bread.
1.3 Yeast taxonomy
Yeast is an immobile single-celled eukaryotic microorganism that lives on plant leaves, fruit
surfaces, soil, and dairy products. Many types of yeast are widely used in the food industry and in
many industries. On the other hand, many types of yeast are involved in the deterioration of food
quality and the development of certain animal and plant diseases. Yeast has the ability to break down
sugars and form ethyl alcohol and carbon dioxide. Based on this property, it is called sugar fungus or
saccharomycete. Yeast is a unique type of micromycete that is able to produce large amounts of
biologically active substances such as proteins and vitamins ( B1, B2, B6, nicotinic acid ) .
Some microorganisms can cause disease in humans and warm-blooded animals, as well as
affect the taste, quality and appearance of food. One of the goals of taxonomic research is to detect
such microorganisms and stop their use.
Yeast is classified into several categories based on its genetic, morphological, and
biochemical properties. Classification differences are determined by the ratio of cells to carbohydrates,
the need for vitamins that affect growth factors, and the form of spores. Most yeast belongs to the
genus Ascomycetes, the genus Endomycetales, and the genus Saccharomyces, which belongs to the
family Saccharomycetaceae. There are 41 species in this Lodder category.
Classifiers use the characteristics of yeast industry, shape, physiology, resistance to ethyl alcohol,
clarity of the nutrient medium at the end of oxidation, optimal growth temperature, sugar oxidation

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activity of galactose, mellibiosis, raffinose, Sacch.cerevisiae and lower oxidation including research
and classification.
Classifiers have classified brewer's yeast in the 1980s as Sacch.cerevisiae in terms of DNA properties,
and have used Saccharomyces yeast for oxidation.
As yeast is a single-celled, micelle-free fungus, it is classified as a fungus and divided into four
categories that do not have a specific classification unit. Yeasts were divided into three groups.
These include:
 Askomitsyet (Ascomycetes)
 Basidiomycetes (B asidiomycetes)
 Blastomycetes ( Blastomycetes ), Deuteromycet 9 (Deuteromycetes) or imperfect fungi (fungi
imperfecti) .
There are 2 families, 4 subfamilies, 46 genus and 234 species in the Ascomycotina group, 4
families, 13 genus and 43 species in the Basidiomycotina group, 2 families, 23 genus and 327 species
in the Deuteromycotina group. This abyss is based on the characteristics of yeast spores and vegetative
reproduction. In addition, chemotaxonomic and molecular biology methods are widely used in
taxonomic studies. Chemotaxom tests are used to determine the structure of ubiquinone and the
presence of xylose in the cell wall. The ubiquinone structure is used to differentiate between groups,
with O6, O7, and O8 being used to determine ascomycetes, and O9 and O10 being used to determine
chromosome composition, DNA homology, and G-C ratios in basidomycetes. J.A. Barnett (2000)
classified yeast as follows in his ‘Determination of Yeast Properties’. Fungi ( Kingdom )
 Ascomycota ( Division )
 Hemiascomycetes ( Class)
 Sacharomycetes (Order )
 Saccharomycetaceae ( Family )
 Saccharomyces ( Genus )
According to this classification, yeast is included in 6 classes, 9 order, 20 families, 96 genus and 678
species of Ascomycota 2 division of fungi kingdom.
More than 180 yeast cultures were isolated from the Mongolian biosphere, 81 cultures were
identified, classified into 3 classes, 3 order, 4 families, 9 genus and 43 species and then a database of
yeast was created. (“Microbiological processing and use of secondary raw materials in the food
industry” UB. Dis 2005: 11,18,23)

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1.4 Conditions for yeast multiplication, growth and culture
Most yeasts reproduce by budding, and a small number of yeasts divide and multiply. Once
the budding process is complete, the daughter cell continues to multiply without separating from the
mother cell. But most are easily separated from each other. Breeding shape and coal - by brewing
yeast with water, depending classified as genuine and fake. Yeast begins with the formation of spores
and growth bulbs sexually and asexually. The stem cell's genetic material (DNA) is replicated, part of
which is transferred to the tuber, and the tuber gradually enlarges to form a wall separating it from the
stem cell. As the wall between the mother and the stem cell narrows, the stem cell breaks down and
multiplies on its own. At the point where it is separated from the stem cells, spots appear and no more
buds form. In the mother cell, a single bud (polar bud) and a pair of buds (bipolar bud) are formed and
several (many) buds are formed. Oval, oblong, and round-shaped yeasts are propagated by buds,
which last an average of two hours under favorable environmental conditions. Once the budding
process is complete, there are cases of false mycelium forming and proliferating due to inseparability
from the stem cells and stem cells. Natural rod-shaped yeast divides and multiplies. At this point, a
transverse septum is formed in the cell. Such a phenomenon is rare, observed only in yeast with bomb-
shaped cells. [2-10]
Spore-forming yeast cells are oval, round, sprouting cells elongated, immobile, gram-positive,
do not form capsules, spores are located in cells 2,4,8. Spores create ( asporyegyen ) yeast cells of
young mitsyeli shaped cell cultures 2-5 * reaches of 3.0-7.5 micron size, mature blood cells, 14-16
microns.
Figure2 . Sprouting yeast and sprouting scars
On the surface of the yeast medium grows yellowish, sometimes bright white, with smooth
edges and round colonies. The cells are round, oval, some oblong, and are well stained with methyl
blue dye. Yeast can grow in facultative anaerobic and acidic environments, and it is important to
determine its ability to form spores and pseudo membranes, using Saburo's medium containing
glucose and peptone, usually to form translucent, smooth, medium-sized colonies. Ammonium salts,
amino acids and peptides are used as sources of yeast carbon. Decomposes lactose and sucrose from
carbohydrates. ( Asanov 1980: fodder reserves 1986 )

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The ideal fermentation temperature is 28-30 ℃ and the nutrient absorption is best at 30 ℃.
However, it does not die at low temperatures, and pressure is important for its viability even when
thawed. At pressures above 2800 kgcm2, the yeast dies instantly. Under unfavorable environmental
conditions, yeast proliferation is slowed down, resulting in the formation of stagnant cells
(arthrospores). Artospores are cells with a thick, dense layer of two layers that contain resources such
as glycogen and fat. They are more resistant to heat and dry matter than vegetative cells, and begin to
germinate like vegetative cells when a favorable environment is created. There are a total of 6,000
species of 96 types of yeast, of which 10 are used in food. Widespread yeast strains in food belong to
the genus Saccharomyces. This type of yeast reproduces in the form of spores and buds.
Saccharomyces differ from each other in basic characteristics such as the intensity of the growth
process.
When the appropriate pH-4.5-5.5 values of the yeast culture medium fluctuate up and down, reaching
4.0 and 7.0, the yield decreases and the quality of the yeast deteriorates. This is explained by changes
in environmental factors such as enzyme activity, metabolism and respiratory rate. It also requires
amino acid metabolism to reduce the biomass yield of yeast. However, at pH 5.8, it is noted that a
large amount of nutrients accumulate in the yeast. At our plant's pH of 5.5-5.8, 24-54% of the protein
and 21-30 g of biomass accumulated. Yeast growth, like other microbial growth curves, takes place in
four stages.
Lag-phase. At the beginning, no cell growth is observed. During this time, the yeast cells adapt to
their environment and the amount of RNA inside the cell increases.
Log-phase. During rapid development, cells multiply geometrically, and biologically active young
cells multiply in the medium. Yeast biomass builds up, and at the end of this phase, nutrients are
depleted, metabolic products accumulate, and growth slows.
Stationary-phase. When stagnation or maturation occurs, the number of new and extinct cells
becomes equal, and the effects of accumulated toxins in the nutrient medium increase, creating an
unfavorable environment.
Extinction phase. At the time of extinction, the yeast cells are able to adapt to the new environment
under conditions of depletion of nutrients and high levels of toxins, which disrupt cell
growth, stagnate growth and reproduction, but do not die completely. From the biological point of
view of yeast cell proliferation and growth, the culture cells used in production go through these stages
and complete the technological process.
Metabolism in yeast cells begins when the cell receives nutrients from the external environment,
and as a result of metabolism, yeast multiplies and biologically active compounds are
synthesized. Nutrients penetrate the entire surface of the cell, and under suitable conditions, the yeast
cells process 30-40 times more nutrients per day than their own weight. Precipitated organic
compounds such as proteins and polysaccharides are converted into simple compounds and then used
as energy and metabolic sources.

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The cytoplasmic membrane is responsible for the inflow and outflow of nutrients. When the
concentration of the substance in the medium is higher than inside the cell, the nutrients penetrate the
cell by simple diffusion and do not require energy. However, when the concentration of the substance
in the environment is lower than in the cell, the transport proteins in the cytoplasmic membrane
penetrate the cell. This is done with the participation of energy. Adsorption of anti-charged substances
on the electrically charged surface of the cell is important for metabolic processes, and depending on
the charge and pH of the cell surface, it is positive in an acidic environment and negative in an alkaline
environment.
Brewer's yeast (sacch.uvarum and sacch.carlbergensis) is rich in purine and pyrimidine bases
of RNA, and its extracts can be used as animal supplements to improve the animal's immune system
and improve its resistance to bacterial infections. Nucleotide-rich beer enhances the immune system,
mainly because these nucleotides alter T-cell activity and activate macrophages and B-cells, as well as
the RNA and nucleotide components derived from these substances have
immunostimulatory ( immune- boosting ) effects on pigs. It is widely used as a supplement to
cows. These nucleotides indirectly affect the hematology and the immune system.
Kluyeromycesmarxianus, a brewer's yeast strain, has the advantage of being relatively high
in flavorings (5 ' -GMP and 5' -IMP). The yeast biomass used in beer production has a bitter aftertaste
and requires additional action to remove.
 Candida utilis
Kingdom: Fungi
Division: Ascomycota
Subdivision: Saccharomycotina
Class: Saccharomycetes
Order: Saccharomycetales
Family: Saccharomycetaceae
Genus: Candida
Species: C. utilis
 Formal features
The Suslo agar is pale white and slightly rounded and oval in agar. Vegetative cells are round or oval
with an average diameter of 10-15 μm. False mycelium is formed by budding and consists of
interconnected cells

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 Culture characteristics
Creating this strain of 4% solids, liquids suslo environment 30 degrees in one day after sedimentation
basin through the radiation and consists of thick pinch bandages after each month. Under unfavorable
conditions, the formation of folds decreases when physiological activity decreases.
 Physiological characteristics
Used and fermented by oxidation of glucose, galactose, sucrose and maltose. Trealose raffinose,
insulin, xylose, arabinose, rhamnose, ethanol, glycerin, mannitol sorbitol, milk, amber and citric acid
soluble starch are used as sources of carbon. It is unique in that it uses nitrogen compounds such as
ammonium sulfate, asparagine, urine, and peptone as feed.
 Features of technology
This strain is nitrogen-deficient and has the ability to accumulate large amounts of rapidly multiplying
biomass in acidic pH = 3.2-4.2 (alcohol slag). This yeast is used in the production of biological
proteins, animal feed, and flavoring in the food industry.
 Candida tropicalis
 Formal features
Suslo agar forms a pale white, round, well-radiated folded colony. Vegetative cells are round and
oval, 11 * 15 microns in size. False mycelium is formed by budding and consists of interconnected
cells.
 Culture characteristics
This strain radiates through the walls of the sedimentary tank at the bottom of the liquid
after 30 days in wort with 4% dry matter, and after a month it forms a thick follicular color. The shape
Kingdom: Fungi
Division: Ascomycota
Subdivision: Saccharomycotina
Class: Saccharomycetes
Order: Saccharomycetales
Family: Saccharomycetaceae
Genus: Candida
Species: C. tropicalis
Biological name: Candida tropicalis

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is ring-shaped, but convex on the outside, the structure is homogeneous, and the folds are unique
and curved from other known strains. Under unfavorable conditions, the formation of folds decreases
as the activity of the physiology decreases.
 Physiological characteristics
Used and fermented by oxidation of glucose, gaseous, sucrose and maltose. Trealose raffinose,
insulin, xylose, arabinose, rhamnose, ethanol, glycerin, mannitol sorbitol, milk, amber and citric acid
soluble starch are used as sources of carbon. It is unique in that it uses nitrogen compounds such as
ammonium sulfate, asparticin, urine, and peptone as feed.
 Features of technology
This strain diet deficient in nitrogen, pH = 3.2-4.2 medium / alcohol slag / ability to accumulate
large amounts of biomass fast times. Under such conditions, the biomass yield is 30-40%. Biomass is
40-55% in an alcohol slag with 0.5% malt decomposition and 0.5% malt sugar extract. Under
favorable growing conditions, the cells are branched and chained buds are intensive. Under conditions
of lack of nutrients and insufficient ventilation, the cells branch out and sprout and germinate
intensively. In the absence of nutrients and insufficient ventilation, the cells separate from each other
and the false mycelial fibers separate. The total protein is 40-50%.
Carbohydrate utilization, physiological and biochemical properties of yeast cultures
Table: 2
Culture
Ability to use hydrocarbon resources
Glucose
Galactose
Xylose
D
-Arabinosis
Sucrose
Maltose
Mellibiosis
Lactose
Fructose
Cellubiosis
Refinement
Insulin
Ethanol
Glycerin
Dulcite
Mannitol
Sorbitol
Lactic
acid
Personal
acid
Citric
acid
Acetic
acid
Tartaric
acid
C.tropicalis + + + - + ± - - - + - - + ± + - +
C.utilis + + + + + - - - + - - - - - + - +

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Biochemical properties Table3
Culture
Ability to ferment sugar
Use of nitrogen
compounds
Urease
activity
Ability
to
form
starch-
like
compounds
Acid
formation
ability
Glucose
Fructose
Galactose
Sucrose
Mellibiosis
Lactose
Maltose
(NH
4
)
2
SO
4
KNO
3
NaNO
2
C.tropicalis + + + - + + - -
C.utilis + + + - - + + + - + - -
Note: + used ± moderately used - not used

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Studied
 The first experiments were performed in Germany to grow fodder on molasses to produce feed
protein.
 Production in 1915 produced 10,000 tons of feed protein. World War I was a time of
economic hardship, and production was halted due to a lack of molasses reserves.
 From the mid-1930s onwards, wood yeast, sulfite, and alcohol were used to make feed yeast.
 In 1935, the Council began using straw and pre- processed sulfuric acid as raw materials in the
Union.
 In the mid-1960s, the French scientist F. Champagne proposed the use of liquid n-alkanes as a
"dirty" raw material in the production of feed protein, which was soon introduced into production.
 V.N Naumenko and S.N Marinskaya (1980) studied that after neutralization of sawdust
hydrolyzate, it is possible to divide it into two parts, grow fodder yeast in one part and mix it with the
other part, steam it and produce hydrocarbon feed. Various hydrolysates have also been widely used,
and experiments have shown that these raw materials have a positive effect on capital growth.
 Our country's light food industry produces a large amount of waste every year, and the issue of
its use has not been fully resolved. Studies have shown that alcohol slag from the plant contains 91-
93 % water, 1.08 % soluble carbon, 5-9 % dry matter, 0.26 % starch and 0.18 % fiber.
 Some chemical parameters of wheat slag in Ulaanbaatar distillery ( D.Tserendulam, 1978 )
were compared with barley and potato slag in Russian distillery. Nitrogen 0.154 in alcohol slag % is
0.11% of the protein compound and 0.095 % of not protein accounted for low molecular weight
compounds and minerals. In other words, alcohol slag is low in nitrogen and phosphate in the form of
feed yeast. Therefore, in order to create the right ratio of hydrocarbons, nitrogen and phosphorus
compounds that can be used in alcohol slag and yeast, it was effective to prepare a nutrient medium by
adding ammonium sulfate, di-ammonium phosphate, superphosphate, hydrogen phosphate, potassium
and urea.
 One of the main conditions for the growth of strains commonly used in the production of feed
yeast is the acidity of the environment and the acidity of the alcohol slag is 1.8 pH = 4.5-5, which fully
meets the above requirements. However, in order to increase the amount of nitrogen and phosphorus
that can be used by the yeast in the slag and the amount of biomass accumulation, it is necessary to
prepare a nutrient medium by adding 0.5g/l of sulphatammonium and diammonium
phosphate.

17. 17
 In our country, livestock breeding is growing year by year, and the amount of protein to be
absorbed by it is increasing accordingly and thousands of tons of feed protein is needed to cover the
shortage of pig and poultry protein. The use of feed yeast in animal husbandry has a high economic
effect on increasing the yield per animal. The amount of biologically active substances in the
composition of animal feed decreases sharply in the spring and winter, when the grass and
cracks. Therefore, enriching animal feed with protein and vitamins is one of the most important issues
to increase its nutritional value and digestibility. Scientists in the field of biotechnology in our country
have conducted research to create raw material resources and highly active strains for the production
of feed yeast using waste from food production (secondary products).
 A.I.Zilberg and A.Leites argue that in addition to "useful" yeasts in beer juice, there are
"useless" and even harmful yeasts, and that the use of pure yeast cultures in production is
essential. For example, Russian scientist ED Faradniva and EV Eroshkina studied the fermentation
activity, flocculation, and reproduction of four strains of yeast, N, 129 P, B, and 8a ( M ) , and
conducted microscopic studies. In the study, the highest fermentation activity of these strains was
8a (M), with 37.1% of the strain activity on the H strain, 129 P on the 25.9%, and 16.6% on the B
strain. The H strain of yeast has a high settling capacity, i.e. the beer fermented with it has a good
clarity, which is a low - oxidation, high-activity yeast belonging to the strain of Sacch.calsbergensis. It
has been studied that the basic properties of yeast technology are preserved when fermenting malt
juice with 12% N dry matter in various industrially circulating yeasts. During fermentation, the H
strain is 24.6-30.2% higher than the 8a (M) strain. The precipitation capacity of the N strain is 10.15%
higher than that of the 8a (M) strain, just as the fermentation activity is different. The malt juice with
11% dry matter was fermented with 4-5 rotations of yeast in 6-8 until 4.3-4.6% dry matter. At this
time, the initial concentration of yeast cells is 20 million / ml, when the H strain is used, the main
fermentation lasts 4 days, and the B and 129P strains each last 5 days, and the 8a ( M ) strain lasts 7
days. The strain contains 92.4 million / ml of yeast cells for 3 days at B-3.5 days, and 80.1 million / ml
of yeast cells at the end of fermentation. At the end of fermentation, the young beer temperature is 1-
20C and kept at 0.04-0.05 MPa for 21 days.
 In 1970, the Institute of Biology of the Mongolian Academy of Sciences, under the direction of
Academician T.Puntsa, conducted an ecological study of the distribution and distribution patterns of
microorganisms in Mongolia under the supervision of Academician T.Puntsa.
 Resolution of the Council of Ministers for the Development of Microbiological Science in
Mongolia was issued on August 15, 1974. In order to implement the resolution, the staff of the
Microbiology Sector under the leadership of Dr. T.Puntsa identified biologically active actinomycetes,
yeasts, algae and bacteria from microorganisms in the Mongolian biosphere. Intensive research into
the production of highly active industrial strains, feed proteins, non-synthetic amino acids, enzyme
preparations, and feed antibiotics has laid the foundation for the development of industrial
microbiology. The study was conducted by B.Tsetseg, T.Narantsetseg, G. Onkhor, D.Jijigkhen,

18. 18
Ts.Dalanbayar, D.Tserendulam, J.Dugarjav, O.Batmunkh, N.Buyankhishig and D.Sanjdorj started
it. The microbiology sector established a biopreparation plant at the pharmaceutical plant in 1978 to
produce protein preparations of the Candida tropicalis strain, which increased protein and
vitamin production, and tested milk yields on dairy farms in Batsumber and Gatsuurt, increasing milk
yields by an average of 1.3 liters per day. In addition, yeast cultures have been isolated from the
Mongolian biosphere, a yeast pool has been established, and some culture species have been identified.
 Various substrates are used in the production of microbial products, as they can be processed
into natural raw materials, food and agricultural wastes, and some secondary products, used as a
source of hydrocarbons, and microbiological processes. There are almost no natural compounds that
are not used in the production of biopreparations. The main source of production is plant biomass
with 70 % hydrocarbons. Therefore, the researcher will give detailed information about the waste beer
and alcohol slag from the fermentation industry.
 Grain malt, a waste from the brewery, is high in hemicellulose, pentose, and humic substances,
which can be broken down to produce a nutrient protein. This decomposition of 40 % of pyentoz,
30 % salt contains arabinozyg for feed supyerfosfat and sulphate using ammonia.
 Local strains of Candida tro p icalis of tyekhnoln cultures alcoholic slag
environment ie processed ogi, 1978 International Standard ( Ust 2600-78 ) approval, " microorganisms
strains used in the production of animal protein feed " ( №194,1981 ) certificate from the
invention ( T .Puntsag, Ts.Dalanbayar, D.Jijighen, D.Tserendulam ) . D pirtiin shaarand
yeast incubation residue absorbed protein level increased by 40% and 2.3 times the nutritional quality
of fermentation, 1 kg of feed yeast feed unit was equal to 1.14. Unfortunately, the plant was shut down
in 1981 due to a shortage of raw materials at the APU plant in Ulaanbaatar . ( Dalanbayar Ts.,
Tserendulam D 1976-1980 )
 K back otinoid niilegjüülegch yeast taxonomy conduct biological research, a discipline to
breeding, raising to establish and maintain media environment suitable for culturing and karotint pills
make chicken feed ingredients effectively all tested as a supplement while Tanner kariotint pill
production technology and standards developed ( Ts.Dalanbayar , D.Tserendulam, M.Maidar,
D.Bayarlkhagva, G.Dorj ) .
 To begin the study of protein yeast, we first collected and compiled data on waste raw material
resources, unit prices, and prospects for use from domestic food industries. The selection of strains and
raw materials for the production of protein preparations and the selection of some chemical parameters
have been identified.
 Establish a research laboratory at the APU distillery, conduct yeast research, produce microbial
protein and vitamin biopreparations, collect information on its importance and economic benefits, and

19. 19
use the compound in alcohol slag as a feed to grow a breeding yeast strain Candida tropicalis .
Experimental research on the production of animal feed by microbiological protein extraction of
alcohol slag ( Ts. Dalanbayar, D. Chuluuntsetseg, D. Tserendulam, 1977 )
 The success of the fermentation process in beer production depends to a large extent on the
correct selection of pure culture strains. This is because the growth of a properly selected pure culture
is rapid. This rapid growth minimizes the growth of foreign microorganisms.
 Food additives derived from cultured strains of brewer's yeast sacch.uvarum,
sacch.carlbergensis S -1 contain 55.5 0.09% protein, 2.21% fat, 10.7% ash,
thiamine ( B1 ) 21.9 μg /g, pyrodoxin ( B6 ) 5.2 μg /g. No toxic elements have been
detected. Therefore, the strain fully meets the basic requirements for use in the food
industry. [ D.Tserendulam,2005 ]

20. 20
Scheme 1 shows the technological scheme of beer production. Scheme: 1
Scheme of beer production technology
Malt raw material Cleanse Waste
Mill
To prepare solution
Water, vapor Condensate
Steam,
cruciferous
/table, cocoons,
extracts/
Filter the solution
Hot precipitation
Precipitate the liquid
Condensate
Slag
Boil the solution and
add flavor to sterilize
aroma
Cooling agent
Precipitated yeast
Cooling,cooling
agent
Fermentation
Cooling
Transfer the finished
beer to the warehouse
Carbon dioxide
Parking in barrels and
bottles
Filter
Precipitated
Construct

21. 21
Alcohol slag
It is a brownish-brown liquid with a sour taste, odor, and 5-8 % dry matter. The slag has the
following chemical composition. The distillate of the distillery has a positive effect on animal feed, but
in order to increase its nutritional value and use it properly, it is used as a breeding ground for
microbiological fodder. Due to the high acidity of the distillery /pH=3.6-4.2/, long-term administration
to animals often leads to changes in stomach acid and acidosis.
Alcohol slag is given to animals in the usual and dried form. The nutrient content of slag
varies depending on the type of raw material. Alcohol slag is usually fed to compost cows and can
give up to 30 liters per day. In addition, milking cows are given 10-15 liters per day and 12-18 liters
per working horse. Slag is easily fermented and degraded during normal storage, so it should be stored
as soon as it is produced, fed to animals or dried with special equipment.
The following products can be made using distilleries.
These include:
 Animal feed
A- Direct drying
B- Preparation of fortified animal feed by growing protein-producing yeast. It is more profitable. Fat,
fiber, unfermented sugars and starches make up 60 % of the dry matter in the slag, which is converted
into yeast biomass during the growth of the yeast. It is a perfect quality feed with protein and amino
acids and the price will increase according to the quality. In order to make the fodder preparation
process cheaper and of better quality, mixing sieves and molds, which are wastes of the flour factory,
reduces energy consumption during drying.
 Biofertilizer: stored in the microbial synthesis laboratory
The active microorganisms can be cultured in alcohol slag to make biological bio fertilizers.
 Biogas

22. 22
Chemical composition of alcohol slag Table3
Specifications Description
Appearance Liquid with a distinctive sour smell
Color Brown
Dry matter 5-8 %
pH 3.6-4.2
General nitrogen 0.12-0.22
Lactic acid 0.25-0.66
Acetic acid 0.05-0.1
Technical alcohol: Semi-processed waste alcohol from alcohol production. Can be used as follows.
These include:
 Paint production
 Manufacture of detergents
 Fuel octane booster
 Denaturation
 Protein-producing yeasts can be used to make animal feed supplements.

23. 23
Beer’s slag
It is a grain residue from the brewing of beer. Beer slag is brownish-yellow in color and has a
pleasant aroma. The following products can be produced using beer slag, which is a waste from beer
production.
These include:
 Make animal feed supplements
A- Direct drying for sale [11]
B-protein-enriched yeast-enriched feed [12]
 Dry beer slag for the production of high-porosity bricks [13]
 Environment for growing mushrooms for food. ( Champignon, oyster )
 Generate energy. Biogas, pyrolysis and direct combustion using beer slag
Energy is generated in three ways, most commonly by direct combustion of biogas, which can cover
up to 60 % of industrial energy consumption, depending on the technological scheme. [14]
 Bio fertilization: It is possible to make biological biofuels using active microorganisms stored
in microbial laboratories.
 Xylitol: E967.
Xylitol is a sugar substitute, similar to sucrose in its sweet taste, twice as much as sorbitol. Xylitol
is used by people with diabetes and overweight as a sugar substitute in their diet. It is used in food
production as a stabilizer and emulsifier, and is used in a variety of food products, including gum. 1
xylitol costs an average of 10,000. [15]
 Using dried beer slag, the “ flour ” is obtained and baked
It is suitable to make a product with 10-15 % of white flour from the flour extracted from brewing
beer. Slag flour contains protein, cellulose, trace elements, fatty acids, vitamins E and D. In the case of
the use of slag flour, the water absorption of the dough increases and less flour is used to produce the
same product in quantitative terms than ordinary white flour. Products made from slag flour are almost
indistinguishable from ordinary products and contain more protein and minerals. It is most
economically profitable for countries that import flour from abroad. Liquid beer can be used to make
bread. [ 16 ]

24. 24
The chemical composition of beer depends on many factors, such as the raw materials used and the
brewing regime of the beer, so it is not constant.
Beerassets:
Beer waste can be used as a microbial feed, and the autolysate is an important addition to the
feed as it is rich in biologically active substances and easily absorbed compounds. The following
products can be made using waste products from the fermentation of beer production.
These include:
 Extraction of sodium glutamate ( E621, MSG) . Sodium glutamate is the taste of the tongue
It activates sensory receptors and intensifies the taste of food. The taste of glutamate is called
"umami". Kikunae Ikeda first developed sodium glutamate in 1907 and sold it under the name
"adzinomoto" or "taste essence". Sodium glutamate is often used as a seasoning in meat and meat
products. Included in sausages and ready-made soups. [17]
 Yeast auto lysate
 Protein, vitamin drinks, seeds, tablets
 Laboratory environment
 Food flavors.
Yeast auto lysate is the concentration of water-soluble compounds in the yeast cell. Yeast cells contain
proteins, carbohydrates, fats, vitamins, and minerals, and can be used to make drinks and pills that
enhance the human immune system, compensate for protein and various vitamin deficiencies. Yeast
cell breakdown is rich in nutrients, so it can be used by research and medical institutions to prepare the
necessary nutrient medium. The waste material can be broken down by special enzymes to produce
products high in 5 ' -nucleotides. It usually produces 5 ' -guanidine monophosphate and 5-
inosine monophosphate. Of these, 5”-inosine-monophosphate-containing yeast has an excellent
flavoring effect. This flavor has the ability to suppress acidic and bitter tastes. [3-1]
 Animal feed
The fermentation plant's waste alcohol, beer slag, and waste assets will be microbiologically recycled
to produce a variety of products, including feed and protein, to create a waste-free plant.

25. 25
CHAPTER TWO.
Research materials and methodology
Research material: Waste slag from APU LLC Brewery. C andida tropicalis and C andida
utilis strains were used in the Microbial Synthesis Laboratory of the Institute of General and
Experimental Biology.
Test site: Experimental and research work was carried out in the Microbial Synthesis
Laboratory of the Institute of General and Experimental Biology of the Mongolian Academy of
Sciences .

26. 26
Methodology used in the study
2.1 Methods of microbiological analysis
1. Defined in accordance with the standard “Method for detection of E. coli (MNS 5367: 2004)”.
1 ml of each dilution of the sample was pre-prepared and injected into a sterile LB liquid medium.
Incubate the inoculated medium at 37 ° C for 24 to 48 hours. If 0.2-0.3 ml of Kovach's reagent is
added to each dilution after incubation in a thermostat, a red color indicates indole or E. coli.
According to the standard “Method for detection of E. coli and E. coli” (MNS 5367: 2004)
2. The total number of microorganisms is determined by counting the number of colonies
grown in Nutrient Agar solids. In doing so, Nutrient Agar solid medium in sterile petri dishes on
him 10 3 10 4 such dilutions drops sterile wedge of 0.1 ml each. Distribute evenly on the surface of
the culture medium (spatula) and incubate at 37 ° C for 24 to 48 h. After removal from the
thermostat, count the total number of microbial colonies growing on the surface of the petri dish.
3. Lactic acid bacteria colonies grown in MRS solid medium no way to count the
number determined. In an MRS solid medium packaged in a 15 ml sterile petri dish , aspirate 0.1
ml of each dilute at 10 3 , 10 4 with a sterile nozzle, distribute it evenly with a spatula, and incubate
at 37 ° C for 72 h. After removing from the thermostat, count the number of lactic acid bacterial
colonies growing on the surface of the petri dish.
4. The number of yeasts is determined by growing the YPD solid medium and counting the
number of colonies. In a sterile petri dish , add 0.1 ml of each diluent to the 10 3 , 10 4 of the sample
on a YPD solid medium packed in 15 ml droplets and distribute with a spatula until evenly
distributed. After incubating in a thermostat at 28 ° C for 24 to 48 hours, the total number of yeast
colonies is calculated.
5. The PDA uses a method to count the number of colonies by growing the total number of molds
in a solid medium. In a sterile petri dish, add 0.1 ml of each dilution to 10 3 , 10 4 of the sample on a
PDA solid medium packaged in 15 ml and distribute evenly with a spatula. After incubating at
room temperature for 72 h, the total number of molds is calculated.
6. The number of thermophiles is determined by growing them in Nutrient Agar solids. In a
sterile petri dish, add 0.1 ml of each dilution to 10 3 , 10 4 of the sample on a solid
medium containing 15 ml of Nutrient Agar and distribute evenly with a spatula. Incubate at 55-60 °
C for 24-48 hours in a thermostat. Finally, count the number of colonies of thermophilic bacteria.
7. Lactic acid bacteria colonies grown in a solid environment MRS ytoo ` counting
method.
8. Staphylococcus aureus grown on CHROM agar solids to count the number of colonies ,

27. 27
9. Cl.perfringens was grown in Wilson-Blair environment at 43 ° C to count the number of
colonies.
10. To determine the number of Salmonella, the method of counting the number of colonies grown
in CHROMagar solids.
2.2 Moisture: by drying (MNS 254-5: 87)
Tools used: Technical weights, drying ovens, buckets, desiccators, clamps
Procedure: Stabilize the box, weigh 5 g of the technical sample and place it in a pre-prepared
box. When the oven temperature reaches 130° C, remove the sample box lid and place it in the
oven. After drying the sample in the oven for 40 min, remove the box briefly, cover with a desiccator
and allow cooling for 10 to 15 min. The cooled sample shall be weighed to the nearest 0.01, and the
moisture content shall be calculated by the following formula for the difference between the pre-
drying and post-drying weights.
Formula-1
X=
𝐵−𝐶
𝐵−𝐴
∗ 100
B- Box weight with pre-drying sample, d
C-Weight of the box with the sample after drying, d
A-empty box weight, d
2.3 Lowry's method for protein determination:
This is the most sensitive of the methods for protein determination by combining the biuret
reaction (recognition of peptide bonds) and the fooling reaction (recognition of tyrosine and
tryptophan). Add 6 ml of a mixture of 6 ml of Na2Cu3-CuSO4 to 1 ml of the test solution containing
50 to 250 μg of protein and leave for 10 min at room temperature, then add 0.5 ml of folic acid reagent.
After 30 to 40 min, the Ext (density) was measured at 660 nm (SF-26) and 750 nm (FET or specol)
against the control solution. Make a mixture of 1 ml of water and 5 ml of Na2CO3-CuSO4 with 0.5 ml
of foliate reagent. Standard albumin solution, 250 μg/ml. The calculation was performed using a pre-
formed pure albumin reference curve.
2.4 Keldel method for protein determination: (MNS 3746: 84)
Proteins in yeast biomass were determined by the micro-Keldal method. The principle of the
method is to oxidize the organic matter in the test sample with a strong sulfuric acid in the presence of
an accelerator, and to combine the resulting urea with a weak sulfuric acid to form ammonium

28. 28
sulfate. Ammonium sulfuric acid was reacted with a strong alkali to absorb the released urea into a
weak sulfuric acid, and the excess acid was titrated with an alkali.
The following formula gives the percentage of total nitrogen in the sample.
Formula-2
X=
𝑇1 (𝐴−Б)𝑇2×0,0014г×100
В
X is the amount of nitrogen in the sample, ( % )
0.1 sulfuric acid in the A-receiver, ( ml )
0.1 N sodium hydroxide used for B-titration, ( ml )
B-sample weight, ( d )
T 1 - acid titration interval
T 2 - alkaline titer interval
2.5 Determination of carbohydrate volume.
Bertrand's method is used to determine the amount of carbohydrate. The basis of the method is
to boil the sugar with copper sulfate to reduce the divalent copper to monovalent copper oxide. The
resulting red copper oxide is filtered; the filtered precipitate is reacted with a solution of iron-
ammonium kvass and titrated with a solution of potassium sulfuric acid to determine the amount of
glucose in contact with copper oxide. The ratio of free copper to common sugar is determined from a
special table.
Add 10 ml of water, 20 ml of copper sulfate and 20 ml of sodium hydroxide to 20 ml of the
analyze, boil for 10 min and then cool. The precipitate is filtered through a No. 3 glass sieve into a
Bunsen flask and the resulting copper peroxide solution is pumped out.
Copper oxide precipitates oxidize in air and must always be submerged. After washing,
transfer the sludge from the stopper to a specially washed Bunsen flask, add the ammonium kvass
solution and stir the precipitate with a glass rod until the copper oxide is completely dissolved and
warm water is pumped out. Bunzl flask 0.1n solution KMnO4 solution of 30 seconds elapses
side slight pink color mass titration.
Formula-3
X=(a-b)*K*100*1000*V1 / n*V 2
Of which: a- KMnO4 used for titration of the control
c- KMnoO4 used to titration the subject
K is the titration coefficient
n is the sample

29. 29
V 1 - original sample
V 2 - sample taken for titration
2.6 Method for determination of fat.
Dry the test piece to constant weight, take a sample from it and apply it to the Soxhlet
apparatus with an organic solvent, measure the previous weight and determine the amount lost by
dissolving in the organic solvent. The Soxhlet apparatus consists of an extractor and a refrigerated
distillation flask. Wrap the specimen in a constant weight of filter paper, weigh it, place it in the
apparatus extractor, ether up to 2/3 of the volume of the distillation flask, place it in a 40 ℃ hot water
bath, turn it 5-6 times and distill it. and then dried at 100-105 100 until constant weight. The difference
between the final and initial weights of the bagged sample is calculated by the following formula for
the amount of oil per 100 g of sample.
Formula4
C= (B-A)*100/C
Of which: C-fat content in percent
A is the weight of the sample in a paper bag before distillation
B is the weight of the sample in a paper bag after extraction
C-sample in grams
100- To make 100 percent
2.7 Bertrand's method for determination of sugar:
Determine the total sugar content. The carbohydrate content (C3) is determined by the following
formula.
Formula-5
C3=
90∗[0.0625 ∗𝛼∗𝛿−(𝑏−2)]
𝑎∗(100−𝑊2)
* (100-W1); %
δ- The amount of sugar in the sample reacted with the feline liquid, ml
b is the pentose content in the weighed sample, d
2- Amount of carbohydrate included with enzyme preparation, %
W1 - initial moisture content, %
W2 - Moisture content in the ground part, %
a-is the mass of the yeast ( powder ) , d

30. 30
CHAPTER THREE
Discussion of research results
3.1 For the culture of Candida tropicalize and Candida utile on beer slag, the density of yeast cell
growth is shown in curve 1.
Yeast cell density growth Curve 1
From the above curve, it can be seen that the C.tropicalis strain of yeast cell growth took 8-12 hours to
adapt to the environment, 12-18 hours the period of intensive growth or maximum cell growth density,
18-24 hours the stabilization period, and 24-72 hours the extinction stage. is shown. The C. utilis strain,
on the other hand, has been acclimatized for 8 to 12 hours, intensive growth for 12 to 18 hours,
stabilization for 18 to 24 hours, and extinction for 24 to 72 hours. .tropicalis, C.utilis strains appear to
have the highest (short-term) activity, with cell growth rate occurring at 12–18 h.
3 .1.1 Number of yeast cells Table 5
Culture The number of cells
C.tropicalis 2.46 * 10 8
C.utilis 17.5 * 10 6
When counting the number of cells of C.tropicalis and C.utilis in the growth (log) phase,
C.tropicalis is 2.46*108 and C.utilis is 17.5*106.
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
8 10 12 14 16 18 20 24 48 72
The
number
of
cell,
million/ml
Time (hour)
C.tropicalis
C.utilis

31. 31
3.2 Determination of some chemical components of brewery waste
Some chemical components (protein, carbohydrates, fats, ash) of APU LLC brewery waste (barley)
slag to be used in the study were determined and compared with the researchers' results are shown in
Table6.
Chemical characteristics of beer slag, %
Specifications Protein Carbohydrate Fat Ash
Beer slag of APU LLC 14 10 6 1.6
Dried beer slag (Russian) 23.44 14.3 7.75 2.3
Beer waste residue and enzymes did extensive (Pirkko Forssel) 21.5 20 9 -
The table shows that the beer waste slag contains 14% protein, 10% carbohydrates, 6% fat and ash,
which is close to the results of the researchers.

32. 32
3.3. The amount of yeast cell growth density
Candida tropicalis and Candida utilisom were replanted in oblique agar and then cultured in a flask to
prepare seedlings. In a 500 ml flask, add 100 g of slag + 200 ml of water, add the feed ingredients,
incubate the sterile yeast strain, ferment for 28 h at 72 ° C, and show cell growth in curve 2.
Candida tropicalis, Candida utilis strain cell growth Curve 2
From the above curve, it can be seen that the density of cell growth was cultured by
C.tropicalis at 0-8 hours of acclimatization, 8-18 hours of intensive growth or maximum growth
density, 18-20 hours of stabilization, and 20-72 hours of extinction. Seems to have taken place.
Incubation of C. utilis showed that the adaptation period was 0-8 hours, the period of intensive growth
at 8-14 hours, the period of maximum cell growth rate was stable, the period of stabilization was 18-22
hours, and the period of extinction was 22-72 hours. C.tropicalis and C.utilis were cultured together at
0-6 hours of acclimatization, 6-14 hours of intensive growth or maximum cell growth density, 14-18
hours of stabilization, and 20-72 hours of extinction. I can see. Incubation of yeast strains C.tropicalis,
C.utilis, (U + T) showed the highest cell growth rate (18-22 hours) (short-term).
0
0.5
1
1.5
2
2.5
3
3.5
0 4 6 8 12 14 18 20 22 48 72
Cell
growth
density,nm
Time (hour)
C.tropicalis
C.utilis
U+T

33. 33
3. 4 Researchon the production of protein feed using brewery waste
Table 7 shows the results of culturing Candida tropicalis and Candida utilis, the protein synthesizers of
the microbial fund of the Microbial Synthesis Laboratory of the Institute of General and Experimental
Biology, using APU's brewery waste slag.
Incubation of Candida tropicalis and Candida utilis strains Table: 7
Used raw
materials
Specifications,% Beer slag
Strain
Candida
tropicalis
Candida utilis C.tropicalis , C.utilis
(T + U)
Beer slag
Protein 14 28 25 23.5
Differences in
protein
14 ( 2 ) 11 ( 1.78 ) 9.5 ( 1.6 )
Experiments have shown that beer slag contains 14% protein, while beer slag produces 28% protein
for Candida tropicalis, 25% for Candida utilis, 23.5% for Candida tropicalis and Candida utilis, and
1.6-2 times more protein than dried slag protein. Candida tropicalis was selected as a strain for
supplementary animal feed.Table 8 shows the comparison of the biopreparation of the yeast Candida
tropicalis in beer slag with beer slag.
Chemical characteristics of beer slag, % Table: 8
Specifications Protein Carbohydrate Fat Ash
Beer slag of APU LLC 14 10 6.0 1.6
Bio preparation 28 18 9 2
The table shows that the amount of fat and ash is similar compared to APU's beer slag and the bio
preparations used. But protein 2 has been increased performance, the highest protein content. But the
face of the water is 1.8 are added.

34. 34
General scheme for the production of protein feed bio preparations Scheme: 2
The dry preparation was obtained by drying the biomass of C.tropicalis yeast and some of its
biochemical parameters were determined. The dry preparation contains 28% protein, 20%
carbohydrates, 6.0 fats, 5.7% moisture and 0.38% ash. The use of brewery waste to produce feed
protein has the advantage of solving the problem of supplemental feed, as well as waste-free
production and reducing environmental pollution.
Pure culture
Fermentation (24-48 hours, 28-30 ℃)
Shale environment
Installationonoblique agar
Grow in a flask Incubate in a 500 ml flask in a 100 ml medium on
a shaker to prepare the stock
Pure yeast cultures were grown in a thermostat
by immersion in a tube with oblique agar
Candida tropicalis, Сandida utilis strain
Grow ina nutrientmedium
Seedlingmaterial seedlings
Drying
Packing
Sack
Drying

35. 35
3.5 Some chemical parameters for culturing yeast strains using slag are shown in the diagram
Diagram: 1
The amount of yeast culture cultured using slag
Compared to the control sample, C.tropicalis had twice as much protein, C.utilis had 1.78 times as
much protein, C.tropicalis had 1.8 times as much carbohydrate, and C.utilis had 1.2 times as much
carbohydrate. But the oil ash is generally close.
14
6
10
1.6
28
9
18
2
25
8
15
1.7
23.5
7.7
14.6
1.9
Protein Fat Carbohydrate Ash
Control sample Slag+C.tropicalis Slag+ C.Utilis Slag+ C.tropicalis+C.utilis

36. 36
Figure 2 shows the protein content of the biopreparations obtained by culturing Candida tropicalis and
Candida utilis in the waste slag of APU JSC's brewery compared to other feeds.
Comparison of protein content of protein biopreparations Diagram: 2
The protein content of these feeds is 1-2 times higher.
0
5
10
15
20
25
30
Cocoon
feed
Powderful
mixed
fodder for
dairy cows
Powerful
mixed
fodder for
ruminants
Powerful
compound
feed for
cattle
Cultivated
C.tropicalis
and C.utilis
in beer slag
Cultivated
C.utilis in
beer slag
C.tropicalis
cultured in
beer slag
10
20
15
20
23.5
25
28
Total protein %

37. 37
3.6 Microbiological analysis
Microbiological and hygienic analysis of protein preparations is shown in Table 2.
Table 3. Results of microbiological analysis
№ Test name
Test results
Beer slag Protein preparations
1 Total number of microorganisms 92 * 10 4 14 * 10 4
2 Lactic acid bacteria Undetected Undetected
3 Yeast Undetected Undetected
4 Mold and mildew Undetected Undetected
5 E. coli Undetected Undetected
6 Thermophilic number 15 * 10 4 Undetected
7 Staphylococcus aureus Undetected Undetected
8 Cl.perfringens Undetected Undetected
9 Salmonella, Shigella Undetected Undetected
According to the analysis, the microbiological parameters of E.coli, S.aureus, Salmonella, fungi,
yeast lactic acid bacteria, Cl.perfringes in beer slag are undetectable and the total number of bacteria is
acceptable. E.coli, S.aureus, Salmonella, no fungi were detected in the black protein, and the total
number of bacteria was within acceptable limits.

38. 38
CONCLUSION
1. APU LLC Barley slag is 14% and contains protein. After culturing two types of yeast, Candida
tropicalis 28% (2 times more), Candida utilis 25% (1.78 times more), C.tropicalis, C.utilis (T +). U)
had a protein content of 23.5% (1.6 times), so the strain Candida tropicalis was selected for further
study.
2. The general scheme of technological process for production of protein feed biopreparations has
been developed.
3. Biopreparations obtained by culturing C.tropicalis strain in beer slag contain 28% protein, 18%
carbohydrates, 9% fat, and 2% ash, and the protein content is 1-2 times higher than that of some
animal feeds. Therefore, there is an opportunity to create waste-free production.

39. 39
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APPENDIX
Seedlings grown in Beer waste slag flasks
Preparation of beer slag medium after fermentation on slag

42. 42
Cultivation of yeast in slag
Dried protein slag dried for 550C 48 hours
Powdered protein feed