RU2562146C2 - Method of producing forage protein product on grain raw material fermentolysat - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology and can be used in production of forage protein products. Method includes preparation of grain raw material (bran, substandard grain, fodder flour) by its milling to particles with size 1-20 mcm by means of mill under pressure with shift and preparation of water suspension of grain raw material with concentration of dry substances 15-16% with ratio of grain raw material and water 1:5. Carrying out fermentolysis of grain raw material by means of liquid α-amylase and glucoamylase with formation of glucose to 10-13%. Introduction of nitrogen and phosphorus sources into obtained fermentolysat; as nitrogen and phosphorus sources used are ammonium or phosphate salts or fertilisers, including ammophos, diammophos, nitrophosphate, with obtaining nutritional medium. Introduction into nutritional medium of culture-producent, as such used is strain of yeasts Saccharomyces cerevisiae Y-3585, and continuous growing in multisectional growing apparatus with supply of nutritional medium simultaneously by several sections with further concentration of suspension on vacuum-evaporator and drying. Moisture, released at stages of vacuum-evaporation and drying, is re-used for preparation of water suspension of grain raw material.

EFFECT: invention makes it possible to increase content of raw protein in finished product and simplify method of production of forage protein product.

5 cl, 7 tbl, 9 ex

Description

1. The technical field

The invention relates to the field of biotechnology, in particular, to obtain protein feed products by microbiological synthesis.

2. The level of technology

Currently, there is a trend towards the revival of feed protein plants. The previously existing plants for large-scale production of feed protein used wood hydrolysates (hydrolysis plants with a total capacity of up to 300 thousand tons per year), sulphite liquors (pulp and paper mills-pulp and paper plants with a total capacity of up to 30 thousand tons per year), and post-alcohol distillery grains as raw materials. (Kirovsky plant with a capacity of 10 thousand tons per year) and refined liquid hydrocarbons (plants for the production of protein-vitamin concentrate-BVK with a total capacity of up to 1 million tons per year). Currently, there are only a few workshops of the pulp and paper mill with a capacity of about 5 thousand tons per year.

In connection with the above, the question of restoring the microbiological production of feed protein is relevant and can be achieved by creating new industries or reconstructing existing plants, in particular alcohol plants, using the latest technologies and new types of raw materials.

In recent years, the development of methods and technologies for the production of fodder protein based on the waste of the milling industry (grain, flour, bran) and a mixture of stillage with waste of grain raw materials (RF patent No. 2159287; RF patent No. 2140449; copyright certificate has developed significantly; USSR No. 1595900; USSR copyright certificate No. 1571061; USSR copyright certificate No. 1532580; USSR copyright certificate No. 1507787; RF patent No. 2054881; RF patent No. 2220590, France patent No. 2646435; French patent No. 2646436; author's certificate of the USSR No. 1677057; No. 1742320; No. 1575570 chipboard; patent C A №4379845).

The disadvantages of the known developments are that some of them are suitable only for office conditions, as they require sterile cultivation conditions. In addition, in most methods of producing fodder protein, imperfect yeast-like fungi of the genus Candida, belonging to conditionally pathogenic microorganisms, are used as producer cultures, which does not meet modern increasing sanitary hygiene requirements.

The most appropriate way to ensure the environmental safety of production is to use, as producer crops, perfect mushrooms (yeast) of the genus Saccharomyces (saccharomycetes), which are traditionally used in the baking and alcohol industries.

However, all known strains of saccharomycetes, which are proposed for use in the production of feed biomass in accordance with the above patents, do not meet the requirements for industrial strains of microorganisms. In particular, they require sterile cultivation conditions and are not competitive in non-sterile conditions of continuous continuous growth processes, being replaced by extraneous strains of microorganisms.

The closest analogue (prototype) to the proposed method is the patent of the Russian Federation No. 2220590 "Method for producing a protein feed product based on grain raw materials."

The essence of the known method lies in the fact that the cultivation of microorganisms producing protein feed product is carried out under aeration conditions on a nutrient medium containing bran and / or cereal flour as a carbon source, treated with RPA cavitation apparatus for 20-50 minutes at a hydromodule (ratio raw materials and water) 1: 5-1: 8. As producers of feed protein, a mixture (in various ratios and compositions) of yeast - saccharomycetes Saccharomyces cerevisiae (S.diastaticus) VKPM-Y-1218, the perfect fungus Endomycopsis fibuligera VSB-12 and / or bacteria Acinetobacter calcoaceticus 0-1 with amylolytic enzymes is used as well as imperfect fungi Trichosporon cutaneum VSB-775, Polyporus sguamosus VSB-917, utilizing the metabolic products of yeast, fungi and bacteria, and which make up 20-30% of the total number of microorganisms.

Technical innovations, according to the known method, include a low hydraulic module (1: 5-1: 8), which allows the fermentation process to be carried out at a concentration of solids in the medium of 10-11%; cavitation processing of grain raw materials (at least 20 min), which makes carbohydrates (reducing substances-PB without inversion 0.7-1.3%) and mineral elements of plant materials available for microorganisms; the use of a mixture of microorganisms - producers of protein, in which yeast, perfect fungi and bacteria decompose starch of grain raw materials to carbohydrates while consuming them, providing an increase in biomass; imperfect mushrooms utilize organic acids - products of the metabolism of yeast and bacteria with a simultaneous increase in biomass and a decrease in the acidity of the medium and the final product.

The disadvantage of this method are: low (0.7-1.3%) content of free carbohydrates - reducing substances after cavitation processing of raw materials; the absence of an indication of the optimum value of the hydraulic module (an interval is given within 1: 5-1: 8), which complicates the implementation of this technological technique in the production environment; lack of optimal parameters for heat treatment of grain raw materials, in particular temperature and aging time; the use of several strains of yeast, perfect and imperfect fungi, as well as bacteria that have different taxonomic positions and various, often not matching growth parameters, in particular temperature and pH, as producer culture of the feed protein product.

In connection with the foregoing, the aim of the present invention is to intensify the method of obtaining a feed product due to a more optimal grinding regime of grain raw materials; optimization of the hydromodule index, that is, the optimal selection of the ratio of grain raw materials and water, providing the greatest swelling of the resulting suspension; the introduction of the stages of heat treatment and fermentolysis, providing the maximum decomposition of starch to glucose; use as a producer culture of a non-pathogenic strain of yeast (saccharomycetes) for food purposes, having amylase activity, that is, the ability to assimilate the starch contained in the grain raw materials.

3. Disclosure of invention

The technical result when implementing the inventive method using a strain of yeast Saccharomyces cerevisiae VKPM VSB-193 (Y-3585), which has the ability to grow at high speed on starch, which reduces the growing time of Saccharomyces cerevisiae yeast, increase production, increase crude protein, increase the content of true protein and increase the dominance of the producer culture in the industrial microbial population at the growing stage.

In the closest analogue, the growing time of the producer culture is 8 hours, in the claimed method - 5.5 hours. Product development in the closest analogue is 70 tons / day, in the claimed method - 100 tons / day. The crude protein content in the closest analogue is 43-58.2%, true protein - 32.4%. In the proposed method, the crude protein content is 63%, true protein 40%. In the closest analogue, the culture-producer in the biocenosis of the fermenter is preserved by 70%, in the claimed method - 100%.

The technical result is achieved by optimizing the process parameters selected for the use of a new strain of yeast Saccharomyces cerevisiae VSB-193 (VKPM Y-3585) by:

- introducing the stage of preparation of grain raw materials (bran, grain and dermi) by grinding as much as possible with a mill, the principle of which is based on high pressure and strain shear (VDiDS), with the formation of particles 1-20 microns in size, which accelerates and increases the efficiency of the process fermentolysis (decomposition of starch);

- the use of a new method of preparing a nutrient medium in the form of an aqueous suspension of grain raw materials, which is milled bran, and / or grain, and / or tart, as well as / or flour, with a water ratio of 1: 5 with a concentration of dry substances in the nutrient medium 15- 16%, which allows to improve the stage of swelling of grain raw materials and to exclude the separation stage after the process of growing yeast;

- conducting heat treatment for 1 hour at a temperature of 85 ° C and pH 6.0, combined with the first stage of fermentolysis, by adding a heat-resistant alpha-amylase enzyme to the suspension of grain raw materials, providing partial decomposition of starch to glucose;

- carrying out the 2nd stage of fermentolysis, which is carried out for 1 hour at a temperature of 55 ° C and pH 5.0 using the liquid glucoamylase enzyme, which provides almost complete decomposition of starch to glucose;

- the implementation of the process of continuous cultivation of yeast in a multi-sectional apparatus with a dispersed and simultaneous supply of nutrient medium in separate sections, which allows you to evenly distribute it in the total volume of the apparatus;

- optimizing the technology and reducing the cost of the finished product by saving steam at the stages of vacuum evaporation and drying, which used a closed cycle that allows you to reuse the steam released at the stages of vacuum evaporation and drying, and send it to the stage of preparation of the nutrient medium;

- the use of a non-pathogenic strain of food and feed yeast Saccharomyces cerevisiae VSB-193, which has amylase and glucoamylase activity and is characterized by active growth on a mineral medium with soluble (amylose) and insoluble (amylopectin) starch, as a monoculture producer. The strain is able to grow in a long continuous process of growing in non-sterile conditions. The dominance of the strain in the biocenosis of the industrial apparatus is 100%. The strain was identified on the basis of genetic analysis of the 16S RNA base sequence and was deposited in the All-Russian Collection of Industrial Microorganisms (VKPM) under the number Y-3585.

4. The implementation of the claimed method

The essence of the proposed method lies in the fact that the process of continuous cultivation of the yeast strain Saccharomyces cerevisiae VSB-193 is carried out under non-sterile conditions in a multisection apparatus with aeration and mixing, into which a nutrient medium is continuously and simultaneously fed through several sections.

Preparation of a nutrient medium includes several stages: preparation of grain raw materials; preparation of an aqueous suspension of grain raw materials of a specific hydraulic module; fermentolysis of starch to dextrins and glucose; introducing nitrogen and phosphorus sources into the prepared fermentolizate. As sources of ammonium and phosphorus, ammonium phosphate, ammonium sulfate, potassium phosphate mono- and disubstituted, as well as fertilizers in the form of ammophos or diammophos or nitrophosphate, can be used.

The preparation of grain raw materials consists in grinding bran and grain in a mill with the formation of particles 1-20 microns in size. The grinding of grain raw materials was carried out in a high pressure mill and strain shear. The grinding elements of the mill first compress the particles of raw materials to 10-15 Gigopascals, then a deformation shift between the grinding elements follows, as a result, the compressed particle breaks into small parts.

Studied the composition of wheat flour, characterized by different particle sizes after grinding, which is presented in table No. 1

Table number 1 The chemical composition of commercial and ground wheat flour №№ Raw material component Commercial flour Ground flour pp particle size 40-120 microns particle size 1-20 microns one Fat 2.9 4.3 2 Protein 14,99 16.28 3 Cellulose 4.66 4.7 four Starch 45.44 45.35 5 Ash 3.91 6.72

As can be seen from table No. 1, the chemical composition of the grain raw material does not change much when grinding it. However, the cultivation of yeast using native and ground grain raw materials revealed a significant difference.

Table No. 2 presents the results of the cultivation of yeast Saccharomyces cerevisiae VSB-193 (VKPM Y-3585) on native wheat flour with a particle size of 40-120 μm and flour milled with a particle size of 1-20 μm.

Table number 2 The performance of the cultivation of yeast Saccharomyces cerevisiae VSB-193 (VKPM Y-3585) in periodic and continuous modes №№ pp raw materials App. Vtotal / Vpaб. Accumulation / low tide topping rV outgoing,% rV con.% NE Start,% SV Con.,% Expense ratio one Native wheat flour Biotron 50/10 Periodic 2.6 0.36 4.9 2.6 1.88 2 Native wheat flour Biotron 50/10 Continuous 2.2 0.3 4.9 2.6 1.88 3 Ground wheat flour Biotron 50/10 Periodic 2.0 0.11 2.6 2.0 1,1 four Ground wheat flour Biotron 50/10 Continuous 2,4 0.6 2.75 2.45 1,1

From table No. 2 it follows that the expenditure coefficient, which characterizes the ratio of the mass of the finished product to the mass of the feedstock, is 1.88 when grown on native wheat flour. When grown on flour with wheat flour, the consumption coefficient is 1.1, that is, almost 2 times less. This indicates that with large particle sizes per unit of production, much more grain raw material is expended, which is not economically viable. The cultivation of yeast on flour milled thus increases the productivity of the fermenter. The improvement of technological indicators was confirmed in an industrial environment, which is presented in table No. 3.

Table number 3 The performance of the cultivation of yeast Saccharomyces cerevisiae VSB-193 (VKPM Y-3585) in an industrial environment №№ pp Grain Raw Growing time, hour Growth rate, hour ^-1 Production, t / day. Content
crude protein,% The content of true protein,% The dominance of producer culture,% one Native bran, particle size 120-300 microns 8 0.13 70 41.0 35.3 70 2 Milled bran, particle size 15-20 microns 5.5 0.18 one hundred 43.7 38.1 one hundred

From table No. 3 it is seen that the cultivation of yeast on bran, subjected to grinding with a particle size of 15-20 microns, leads to a reduction in growing time from 8 to 5.5 hours, an increase in growth rate from 0.13 to 0.18 hour ^-1 , an increase in the production of the product from 70 to 100 t / day, an increase in the content of crude protein from 41.0% to 43.7%, an increase in the content of true protein from 35.3 to 38.1%; the dominance of the producer culture increased from 70 to 100%.

The preparation of an aqueous suspension of grain raw materials was carried out by mixing (using a mixing device, for example, a mixer) milled bran and / or grain, and / or turf, as well as / or flour with water in a ratio (hydraulic module) of 1: 5, which ensures the concentration of grain raw materials in a nutrient medium 15-16% and the best swelling of grain raw materials.

The prepared aqueous suspension of grain raw materials is subjected to heat treatment at a temperature of 85 ° C, which proceeds at pH 6.0 and adding to the suspension 0.16 g / kg of grain raw material of the enzyme alpha-amylase - “viscoferm” (manufactured by Denmark, Novozyme company), which is a mixture of xylanase, beta-glucanase, alpha-amylase, cellulase. The duration of the heat treatment stage, combined with the simultaneous partial fermentolysis process (1st stage of fermentolysis), is 1 hour. In this case, the starch is liquefied and partially decomposed to dextrins and glucose. Next, the second stage of fermentolysis is carried out using the enzyme “diazym SSF” (manufactured by Denmark, Danisko), which contains glucoamylase, which decomposes starch to glucose. The enzyme is introduced in an amount of 0.48 g / kg of grain raw materials. Fermentolysis proceeds at pH 5.5 and a temperature of 55 ° C for 1 hour. The content of glucose (reducing substances) in the fermentolizate is up to 13%.

Table No. 4 presents the data of industrial tests on the cultivation of Saccharomyces cerevisiae VKPM Y-3585 yeast on bran and flour milled, as well as subjected to heat treatment and heat treatment combined with fermentolysis.

Table number 4 The performance of the cultivation of yeast Saccharomyces cerevisiae VSB-
193 (VKPM Y-3585) in an industrial environment №№ pp Grain Raw Material Processing option The content of reducing substances,% Crude protein content,% The content of true protein,% one Chopped bran Heat treatment at 80 ° C for 1
hours up to 1 39-40 25-27 2 Chopped bran Heat treatment at 80 ° C for 1 hour + partial fermentolysis with alpha-amylase 1.5-2.0 41-43 30-31 3 Chopped bran Heat treatment at 80 ° C for 1 hour + complete fermentolysis with alpha-amylase and glucoamylase up to 7-8 45-47 35-38 four Ground flour Heat treatment at 80 ° C for 1 hour 2-3 42-44 31-34 5 Ground flour Heat treatment at 80 ° C for 1 hour + partial fermentolysis with alpha-amylase 5-7 45-50 35-37 6 Ground flour Heat treatment at 80 ° C for 1 hour + complete fermentolysis with alpha-amylase and glucoamylase 7-10 55-63 38-40

From table No. 4 it is seen that the introduction, in addition to heat treatment, of the stage of fermentolysis of grain raw materials leads to a deeper decomposition of starch with the formation of a large amount of glucose (reducing substances) from 1 to 13%, an increase in the content of crude protein from 39% to 63%, an increase in the content true protein from 25 to 40%.

Sources of nitrogen and phosphorus in the form of mineral salts or fertilizers are introduced into the prepared fermentolizate, for example, ammonium phosphate mono- or disubstituted, ammophos or diammophos, carbamide, based on the fact that 90 mg of nitrogen and 45 mg of phosphorus are needed for the synthesis of one gram of biomass. Thus, a complete nutrient medium for growing yeast is obtained.

Yeast is grown continuously in an industrial multi-sectional apparatus of pulsed action, while the nutrient medium enters the apparatus in different sections at the same time, which ensures its uniform distribution in the apparatus. The temperature of cultivation is 32-34 ° C and a pH of 4.0-4.5. The growing process is carried out in a continuous mode with a dilution factor of the medium D = 0.12-0.14 hour ^-1 .

After the growing process, the suspension enriched with biomass of yeast is sent from the growing apparatus, bypassing the separation stage, to vacuum evaporation to thicken to a dry matter concentration of 40%. The vapor generated by the vacuum evaporation is condensed and sent to the preparation of a nutrient medium.

After vacuum evaporation, the concentrated suspension is sent to the dryer, dried, and thus the finished product is obtained. The crude protein content in the finished product is 39% -63%, depending on the type of grain raw materials.

As a producer protein culture, a non-pathogenic strain of food and feed yeast Saccharomyces cerevisiae VKPM Y-3585, obtained by selection of cereal biocenosis from raw materials, is used. The yeast strain actively grows on a mineral medium with native soluble and insoluble starch (amylose and amylopectin, respectively). The yeast strain VKPM Y-3585 is able to grow in a wide range of pH 4.0-6.0, the optimal value is 5.0-5.5. The temperature of cultivation is 30-35 ° C, the optimum temperature is 30-32 ° C.

5. Examples of the method

Example 1

Wheat bran was milled in mills of various designs (ball and high pressure with a shift) to a particle size of 1-20 microns, which ensures a stable state of the suspension when mixing bran and water and further optimizes the process of fermentolysis.

The characteristics of ground bran after grinding are presented in table No. 5.

Table number 5 sample micron particle size native bran (control) 300-600 ball mill bran 1-20 shear bran processed in a high pressure mill 1-20

According to the data obtained, the mills of both designs provide grinding of bran with a particle size of 1-20 microns.

The cultivation of yeast Saccharomyces cerevisiae VKPM Y-3585 with amylolytic activity was carried out on a mineral medium, pH 5.5 with 0.5% bran native (particle size 300-600) and ground (particle size 1-20) in flasks on a rocking chair 180 rpm at a temperature of 30 ° C for 20 hours. Growth activity was assessed by the change in pH after the growing process, which decreases more intensively, the higher the growth activity of the culture and the increase in biomass is higher. So, when using native (non-ground) bran as a carbon source, the pH of the culture suspension was 5.0; when using ground bran 3,5.

The composition of the mineral medium (g / l): ammonium sulfate 5; monosubstituted potassium phosphate 0.85; potassium phosphate disubstituted 0.15; sodium chloride 0.1; CaCl × 2H ₂ O - 0.1; MgSO ₄ × 7H ₂ O - 0.1; yeast extract 0.1.

Example 2

Wheat bran was ground in a mill to a particle size in the range of 1-20 microns and used to prepare an aqueous suspension at various values of the hydraulic module 1: 9, 1: 5 and 1: 3, that is, at different ratios of bran and water. Got suspensions with a concentration of bran by dry weight of 10, 16.5 and 25%, respectively. Further, aqueous suspensions of bran were held for 30 and 60 minutes on a rocking chair at a temperature of 50 ° C and stirring at 100 rpm. The state of the suspension was evaluated visually and by measuring the size of the supernatant. The content of reducing substances (glucose) was determined by the Bertrand method. The characteristics of the suspensions are presented in table No. 6.

Table number 6 option bran, g water ml water module, the ratio of bran to water conc. bran,% suspension state; the size of the annoyance. liquid mm pH temp. ° C rV,% after 30 min rV,% after 60 min one 10 90 1: 9 10 sustainable (in the form of jelly); 2 mm 6.1-6.4 fifty 0.33 0.4 2 twenty one hundred 1: 5 16.6 sustainable (in the form of jelly); out 6.1-6.4 fifty 0.7 0.7 3 thirty 90 1: 3 25 sustainable (as 6.1-6.4 fifty 0.42 0.7 thick dough); out

According to the data obtained, the most technologically advanced is a suspension with a 1: 5 hydromodule, in which a stable state of the suspension is achieved without the formation of a supernatant or its minimum value and a glucose content of 0.7% after 60 minutes at 50 ° C due to fermentolysis with its own enzymes, available in bran. Hydraulic modules 1: 9 and 1: 3 are low-tech. So, with a 1: 9 hydromodule, few reducing substances are formed (0.4%), with a high concentration of bran in the suspension (1: 3 hydromodule), the glucose content does not exceed 0.7%.

In addition, the stage of swelling is the first important stage of the fermentolysis process, providing acceleration and depth of decomposition of starch to glucose.

Example 3

A suspension of wheat flour with a 1: 5 hydromodule was subjected to heat treatment and preliminary fermentolysis (stage 1). Heat treatment and fermentolysis were carried out in a 50 L Biotek apparatus. To do this, with stirring at 500 rpm, 30 l of water and 6 kg of flour were added (hydraulic unit 1: 5). Received the concentration of flour in suspension of 15.0% by dry weight. Next, heating was carried out to 85 ° C, pH 6.0 was adjusted, and ViscoFerm liquid enzyme (manufactured by Novozyme, Denmark) was added in an amount of 1.0 ml (0.16 g per 1 kg of grain feed enzyme consumption) and kept within 1 hour. The glucose content at the end of the heat treatment processes with simultaneous partial fermentolysis was 2.8%. Next, the final process of fermentolysis (stage 2) was carried out. To do this, the temperature was lowered to 55 ° C, pH 5.0 was set, and the Diazim SSF liquid enzyme (manufactured by Danisco, Denmark) was added in an amount of 3.0 ml (0.48 g enzyme consumption per 1 kg of grain raw materials) and kept for 1 hour. The glucose content at the end of the fermentolysis process was 13.3%.

Example 4

The process of growing yeast Saccharomyces cerevisiae VKPM Y-3585 was carried out in a two-section pulsed apparatus, in which mixing was carried out by pumping the suspension from one section to another using the air that was supplied to the apparatus. The volume of the apparatus was 60 liters with a working volume of 36 liters. Water was added to the apparatus in an amount of 28 l, 6 l of flour fermentolizate prepared according to Example 3. The temperature was set at 32 ° C and pH 4.5, then 25.5 g of ammonium sulfate and 12.2 g of potassium phosphate monosubstituted were added. Then, 2 L of inoculum biomass was introduced into the apparatus in the form of a suspension of a pure culture of the yeast strain Saccharomyces cerevisiae VKPM Y-3585, previously grown in rocking flasks on a liquid wort. Cultivation was carried out by a detachable-refilling method while automatically maintaining pH and temperature. The results of the growing process are presented in table No. 7.

Table number 7 PB (glucose),% initial RV (glucose),% at the end of the process SV source.% SV at the end of the process,% The selection of the suspension, l Topping up fermentolizate, l Consumption ratio, SV source / SV end 2.0 0.11 2,8 2.6 - - 1,1 2.6 0.36 4.9 2.6 10 10 1.9 2,4 0.21 3,5 3.0 10 10 1,2 2,5 0.1 5,6 3.75 10 10 1,5

According to the data obtained, the optimal growing conditions were those in which the solids content (substrate) was 2.8-3.5% and the consumption coefficient was 1.1 and 1.2, respectively.

The suspension was then concentrated by centrifugation, the supernatant was separated, and the compacted precipitate was dried. The crude protein content in the finished product was 63.0%.

Example 5

The process of continuous growth was carried out in an industrial multi-section apparatus for growing microorganisms (according to the patent of the Russian Federation No. 2144952) with a total volume of 90 m ³ with a working volume of 50 m ³ .

The growing process was carried out as follows. The working volume of the apparatus 50 m ^{3 was} filled in several sections of the apparatus with wheat bran fermentolizate prepared according to examples 1, 2, 3, or wheat flour or mixtures thereof in an arbitrary ratio with a solids concentration of 10-15% and glucose of 8-13%. Mineral nutrition sources were introduced into the environment on the basis that 90 mg of nitrogen and 45 mg of phosphorus are required per 1 g of biomass. Ammophos, or diammophos, or nitrophosphate, or ammonium phosphate monosubstituted were used as sources of nitrogen and phosphorus. The dilution factor of the medium (growth rate) was 0.1 hour ^-1 . The culture suspension after cultivation was subjected to vacuum evaporation and then drying on a spray dryer. Productivity was at the level of 8-13 kg / m ³ / hour. The crude protein content in the finished product is in the range of 48.3-50.7%.

Example 6

A closed cycle of water consumption is reproduced under industrial conditions with a productivity of 1 t / h of the finished dry product with a moisture content of 10%.

The need for water for the preparation of 1 ton of nutrient medium with a concentration of 15% of grain raw materials is 850 kg. This suspension enters the industrial apparatus for growing yeast and leaves the apparatus with a concentration of 10-13%, since 5-2% is spent on the respiration of the yeast during its growth. That is, 1-1.3 tons of the suspension entering the vacuum evaporation consists of 100-130 kg of dry product and 900-870 kg of water. After vacuum evaporation, this suspension is concentrated to a solids content of 40%, based on the capacity of modern vacuum plants, that is, it thickens 4 times. Thus, 1 ton of the concentrated suspension consists of 100-130 kg of dry product and 150 kg of water: we divide one ton of the suspension entering the vacuum evaporation with a concentration of 10-13% by 4, we get 250-325 kg. Thus, the released water is 750-720 kg, which is sent to the stage of preparation of the nutrient medium.

The missing 100 kg of water required for the preparation of the nutrient medium is sent from the drying stage. The calculation of the water circulation at the drying stage is as follows: from the vacuum evaporation stage, 1 ton of suspension with a solids concentration of 40% goes to drying, that is, the suspension includes 400 kg of dry substances and 600 kg of water. From the drying stage, the finished product comes out with 10% moisture, that is, 1 ton of the finished product includes 900 kg of dry product and 100 kg of water. Thus, the released water is 500 kg (600 kg-100 kg). 100 kg of water goes to the stage of preparation of the nutrient medium and 400 kg of water goes to other technological needs (washing equipment, communications, salt solutions, etc.).

Thus, the closed cycle of water consumption fully provides the plant with technological water, while the water consumption is 90%, based on the fact that the production of 1 ton of the finished product with a moisture content of 10% requires 9 tons of water. In addition, due to the closed cycle of water consumption, environmental safety of production is ensured.

Example 7

A closed cycle of steam circulation is reproduced under industrial conditions during the operation of a three-body vacuum evaporation, the operation of which requires 2.5 t / h of steam to obtain the finished product with a humidity of 10%.

The steam consumption for obtaining the finished product is the sum of the steam consumption at the stages of vacuum evaporation and drying. At the vacuum evaporation stage, to thicken 1 ton of a 10% suspension coming from the growing apparatus, 750 kg of water is evaporated, which condenses and returns to the stage of preparation of the nutrient medium. At the drying stage, to dry 250 kg of suspension coming from vacuum evaporation, 140 kg of water must be evaporated, which returns as a vapor to vacuum evaporation. Thus, to obtain the finished product with a moisture content of 10%, it is necessary to evaporate 890 kg of water. For the evaporation of 750 kg of water in the vacuum evaporation stage, 250 kg of steam is required; 140 kg of steam are required to evaporate 140 kg of water during the drying phase. The steam supply deficit, taking into account 140 kg of steam returned to the vacuum evaporation, is 110 kg, which is replenished with steam from the boiler room. This amounts to 44% of the consumed steam, and thus steam losses during drying are reduced by the same amount. In addition, due to the closed cycle of steam circulation, environmental safety of production and energy-saving technology are ensured.

Example 8

The yeast strain Saccharomyces cerevisiae VKPM Y-3585 was grown in flasks (800 ml) on a 180 rpm shaker, on a mineral medium with soluble (amylose) and insoluble (amylopectin) starch to detect the presence of amylolytic enzymes capable of decomposing starch to glucose. The composition of the mineral medium (g / l): ammonium sulfate 5; monosubstituted potassium phosphate 0.85; potassium phosphate disubstituted 0.15; sodium chloride 0.1; calcium chloride at 2H ₂ O - 0.1; magnesium sulfate at 7 H ₂ O - 0.1; yeast extract 0.1; pH 5.5-6.0. 1% soluble (amylose) and insoluble (amylopectin) starch was added to the prepared mineral medium. The medium was sterilized at 0.5 atm. for 20 minutes. Cultivation was carried out at a temperature of 32 ° C for 48 hours. Growth activity was evaluated by the optical density of the suspension on a photoelectrocolorimeter (FEK-56M) device with a green filter (wavelength 560 nm), which amounted to 5.6 units. with growth on soluble starch and 4.8 units with growth on insoluble starch.

Thus, the yeast strain Saccharomyces cerevisiae Y-3585 has amylolytic enzymes and can be recommended as a producer culture to obtain a feed product.

Example 9

The yeast strain Saccharomyces cerevisiae VKPM Y-3585 was grown in flasks (800 ml) on a 180 rpm shaker, on a fermentolizate of a mixture of wheat bran (70%) and wheat flour (30%) prepared according to example 3. pH of the nutrient medium 5.5 , temperature 32 ° C, growing time 20 hours. Growth activity was evaluated by the optical density of the suspension on a photoelectrocolorimeter (FEK-56M) device with a green filter (wavelength 560 nm), which amounted to 28.5-31.0 units. for various repetitions.

Thus, the yeast strain Saccharomyces cerevisiae VKPM Y-3585 has amylolytic activity and can be recommended as a producer culture to obtain a feed product.

6. Conclusions

- Optimization of the grinding of grain raw materials with the formation of particles of 1-20 microns accelerates the formation of glucose during the decomposition of starch at the stage of fermentolysis.

- Optimization of the hydraulic module 1: 5 provides the best swelling of the grain raw material and its subsequent liquefaction, significantly accelerating the stage of fermentolysis.

- The introduction of the stage of fermentolysis can increase the yield of reducing substances to 10-13%.

- Increases the yield of crude protein up to 63%.

- The introduction of closed cycles of water and steam consumption provides steam savings of 44% and water by 90%.

- The use of monoculture strain of saccharomycetes yeast.

Claims (5)

1. A method of producing a protein feed product, providing for the preparation of grain raw materials by grinding it to particles with a size of 1-20 microns, preparing an aqueous suspension of grain raw materials with a solids concentration of 15-16% with a ratio of grain raw materials and water 1: 5, carrying out fermentolysis of grain raw materials sequentially using liquid α-amylase and glucoamylase, introducing nitrogen and phosphorus sources into the obtained fermentolizate to obtain a nutrient medium, introducing a producer culture into the nutrient medium, in which lzuyut yeast Saccharomyces cerevisiae strain VKPM Y-3585, continuous growth in a multi-sectional apparatus with feeding the nutrient medium simultaneously in several sections, subsequent concentration of the suspension in vacuum evaporation and drying, and the moisture released in the stages of vacuum evaporation and drying is used to prepare an aqueous suspension of grain raw materials. 2. The method according to claim 1, characterized in that the fermentolysis is carried out to achieve a glucose content of 10-13%. 3. The method according to claim 1, characterized in that the quality of the grain raw materials used flour, chopped bran, tart and grain. 4. The method according to claim 1, characterized in that the sources of nitrogen and phosphorus are ammonium and phosphate salts or fertilizers, including ammophos, diammophos, nitrophos. 5. The method according to claim 4, characterized in that the sources of mineral nutrition are added based on the fact that 1 mg of yeast biomass requires 90 mg of nitrogen and 45 mg of phosphorus.