RU2243678C1 - Method for preparing protein-vitamin fodder - Google Patents

Abstract

FIELD: fodder industry.

SUBSTANCE: invention relates to a method for preparing protein-vitamin fodder that involves solid or liquid waste in production and processing the natural raw (grains, milling waste, post-alcoholic distillery grains, beer pellets, fruit pulps or whey). Enzyme lysates are prepared from solid waste and starch waste. Cobalt salt is added to liquid waste or enzyme lysates. Prepared nutrient medium is used in incubation of lactobacillus and propionibacillus microorganisms taken by the following pairs: Lactobacillus acidophilus 1660/02 with Propionibacterium freudenreichii subsp. shermanii 103/12; or Lactobacillus acidophilus 1660/02 with Propionibacterium acnes 1450/28; or Lactobacillus plantarum 578/25 with Propionibacterium freudenreichii subsp. shermanii 103/12; or Lactobacillus plantarum 578/25 with Propionibacterium acnes 1450/28. This method provides preparing fodder enriched with vitamins and proteins and containing live cells of lactobacillus and propionibacillus microorganisms. Method enriches animal intestine microflora after feeding the prepared fodder to animals. Fodder comprises protective substances (organic acids, enzyme systems) and can be stored as crude form for the prolonged time.

EFFECT: improved preparing method, valuable properties of fodder.

13 cl, 1 tbl, 12 ex

Description

The invention relates to biotechnology, to agriculture, to feed production.

By the beginning of the 21st century, the world protein deficit is estimated at 30-35 million tons per year. The main way to reduce this deficit is to produce biomass using microbiological synthesis.

Grain raw materials - rye, barley, wheat, flour milling waste, including substandard waste, are used for livestock feed or for the preparation of animal feed. However, grain raw materials are not a complete feed due to their low protein content, poor digestibility, and an unfavorable ratio of proteins and carbohydrates. More promising is the production of protein feed by growing microorganisms on grain raw materials.

Valuable raw materials for the production of feed are waste from industries that process natural raw materials, but there is practically no rational disposal of waste. Partially, the waste is sent to livestock feed in raw form, but the duration of their storage is not more than 1 day. Partially they use drying, for example, alcohol stillage, beer grains, but this process is inefficient due to the high consumption of heat and energy resources and the low quality of the dried product (for protein and other useful substances).

A known method of producing protein-vitamin feed by incubating microorganisms in a nutrient medium containing mineral salts and a carbon source in the form of a waste product from the processing of natural raw materials to obtain the target product in the form of biomass of microorganisms (Patent RU No. 2183666, class C 12 F 3/10 , С 12 Р 7/06, А 23 К 1/06, published on June 20, 2002) [1].

As waste products for the processing of natural raw materials in this known method, grain is used, which is crushed, subjected to enzymatic hydrolysis to obtain an enzyme lysate, from which a nutrient medium is prepared for growing Saccaromyces cerevisiae yeast on it to obtain alcohol. The resulting biomass is used as feed.

However, the yeast is grown under aerobic conditions, which requires significant energy consumption for supplying air to the environment for their incubation. To use the resulting biomass in the feed, it is necessary to heat it in order to stop the yeast from functioning. Such processing requires significant energy consumption and is accompanied by the destruction of part of the biologically active components of the feed. This food does not contain living cells that can settle in the gastrointestinal tract of the animal and improve its performance.

A known method of producing protein-vitamin feed by incubating microorganisms on a nutrient medium containing mineral salts and a carbon source in the form of a waste from the production of processing natural raw materials (Patent RU No. 2054881, class A 23 K 1/165, publ. 02.27.96, ) [2].

In this well-known case, ground grain and waste from flour milling, which is subjected to heat treatment, are then used as waste from the production of processing natural raw materials, after which the yeast of the genus Candida and bacteria producing amylolytic enzymes that carry out the enzymatic hydrolysis of ground raw materials and waste from flour milling are co-grown. .

The disadvantage of this known method is the use of yeast of the genus Candida for the production of protein feed, which is a conditionally pathogenic microorganism, which requires an additional thermolysis process and special equipment, as well as a complex system for the treatment of wastewater and air emissions, requires significant amounts of air used. Sanitary and epidemiological rules SanPiN 2.3.2.1078-01 yeast of the genus Candida are included in the list of substances that have a harmful effect on human health.

Significant disadvantages of this method should also include a large consumption of air and heat and energy resources, the need for high-performance, complex and energy-intensive equipment and a significant consumption of auxiliary materials.

A known method of producing protein-vitamin feed by incubating microorganisms in a nutrient medium containing post-alcohol stillage, to obtain biomass, which is mixed with a carbohydrate-containing component (USSR Author's Certificate No. 1500671, class C 12 P 7/06, publ. 1990) [ 3].

As microorganisms in this known case, yeast is used, which requires aeration during their incubation, and the resulting feed is subject to heat treatment, since it should not contain live yeast cells. This leads to a decrease in the biological value of the feed.

A known method of producing protein-vitamin feed by incubating microorganisms in a nutrient medium containing mineral salts and a carbon source in the form of a waste from the production of processing natural raw materials. (Patent RU No. 2159287, class C 12 P 21/00, A 23 K 1/06, published on November 20, 2001) [4].

In this known method, post-alcohol distillery distillery is used as waste, into which starch-containing raw materials are added — waste from grain-flour production. The prepared mixture is subjected to heat treatment in an acidic medium, and the biomass of bacteria Pseudomonas biforme VSB - 644 is grown on the obtained hydrolyzate.

However, the resulting biomass is subjected to heat treatment, which leads to a decrease in its biological value. In the prepared food there are no living cells enriching the intestinal microflora of animals consuming food. The feed is not subject to long-term storage in raw form.

The technical result achieved by the present invention is to obtain food rich in biologically active components and protein, containing protective substances and living cells of microorganisms, enriching the microflora of the gastrointestinal tract of animals, simplifying the process, reducing energy consumption, improving the safety of raw food.

The specified technical result is achieved by the fact that the method of producing protein-vitamin feed consists in incubating microorganisms on a nutrient medium containing mineral salts and a carbon source in the form of waste from the processing of natural raw materials or in the form of a fermentolizate waste to obtain the target product in the form of biomass of incubated microorganisms, in this case, cobalt salt is used as mineral salts, grain, flour milling waste, starch waste, and after-grain are used as production waste mouth bard, beer pellet, fruit squeezed or whey, fermentolizate is prepared from grain, flour waste, starch waste, beer pellet or fruit squeezed, and incubated in a culture medium are cultures of lactic and propionic acid bacteria used in pairs: lactic acid and propionic acid bacteria together or sequentially : first, the lactic acid bacteria are incubated, and then the joint incubation of the lactic and propionic acid bacteria is continued.

It is advisable to incubate pairs of strains of lactic acid and propionic acid bacteria: Lactobacillus acidophilus 1660/02 and Propionibacterium freudenreichii subsp. shermanii 103/12; or Lactobacillus acidophilus 1660/02 and Propionibacterium acnes 1450/28; or Lactobacillus plantarum 578/25 and Propionibacterium freudenreichii subsp. shermanii 103/12; or Lactobacillus plantarum 578/25 and Propionibacterium acnes 1450/28.

Couples of lactic acid and propionic acid bacteria are recommended to be introduced into the nutrient medium in the ratio (0.8-1.2) :( 0.8-1.2) in an amount of 5-20% vol., And incubate with periodic stirring at a temperature of 37- 50 ° C and pH 5.9-6.0.

During sequential incubation, it is advisable to carry out a joint incubation of lactic and propionic acid bacteria after 1.5-6 hours of preliminary incubation of lactic acid bacteria.

The target product with high biological value, enriched by living cells in the active phase of growth, can be obtained by incubating a pair of microorganisms in a nutrient medium for 3-23.9 hours, and the target product with a high protein content - by incubating them for 24-50 hours .

The fermentolysate of grain, flour waste or starch waste can be obtained by treating it with hydrolytic enzymes: first cellulolytic, and then with a mixture of cellulolytic and amylolytic, while the grain and flour milling waste are pre-crushed, the cellulolytic enzyme is introduced stepwise: first, in the amount of 20-22 units / g dry matter of cellulose and maintain the prepared mixture at a temperature of 59-61 ° C and a pH of 6.0-6.5 for 55-65 minutes, then at a temperature of 95-105 ° C for 25-35 minutes to achieve a reduction 3-5% of the substances, after which 8-10 u / g of dry matter of cellulose cellulolytic enzyme and 5-7 u / g of conditional starch of amylolytic enzyme are additionally added and the prepared mixture is kept at a temperature of 45-50 ° С and pH 5.0- 5.5 for 15-30 minutes until the content of reducing substances reaches 7-10% to obtain the fermentolizate.

Grain or flour waste is ground until at least 80% passes through a sieve with 1 mm diameter cells, and an aqueous suspension prepared by mixing the ground waste with water in a ratio of 1: (3.0-3.5) is subjected to enzyme treatment.

To treat starch wastes with enzymes, a 14-16% aqueous suspension is prepared.

As cellulolytic enzymes, β-glucanase, xylanase or cellulase can be used, and as amylolytic enzymes, α-amylase, β-amylase or glucoamylase.

The fermentolizate of grain, flour waste or starch waste can also be obtained by treatment with amylolytic enzymes, while the waste is first treated with 1.4-1.6 u / g of conditional starch of α-amylase for 55-65 min at a temperature of 59-61 ° C and pH 5.8-6.5, then - for 25-35 minutes at a temperature of 95-105 ° C, after which 0.4-0.6 u / g of conditional starch of α-amylase and 5.5- 6.5 units / g of conventional starch glucoamylase and incubated for 20-30 minutes at a temperature of 45-50 ° C and a pH of 5.0-5.5 until reaching a content of reducing substances of 7-10% to obtain an enzyme olisate.

To obtain the fermentolizate from beer grains, it is advisable to prepare its aqueous suspension by dilution with water in a ratio of 1: 1 and treat with amylolytic enzymes: first introduce 1.4-1.6 u / g of conditional starch of α-amylase into it and maintain the mixture at a temperature of 59- 61 ° C and a pH of 5.8-6.0 for 55-65 minutes, and then for 55-65 minutes at 74-76 ° C, after which an additional 0.4-0.6 units / g of conditional α-amylase starch and the mixture can withstand 35-40 minutes at 59-61 ° C, then enter 5.5-6.5 u / g of conditional glucoamylase starch and the mixture withstand 30-60 minutes At a temperature of 45-50 ° C and a pH of 5.0-5.5 until a reducing substance reaches 7-10% to obtain a fermentolizate.

To obtain the fermentolizate from fruit marc, it is recommended to prepare their aqueous suspension in a ratio of 1: 2, add 1.7-1.9 u / l pectolytic or cellulolytic enzyme to it, and keep the resulting mixture for 2-3 hours at a temperature of 45-50 ° C to achieve a content of reducing substances of 10-15% to obtain a fermentolizate.

Pectin esterase or polygalacturonase can be used as pectolytic enzymes, and cellulase as a cellulolytic enzyme.

The essence of the proposed method consists in the development of waste-free production technology, which provides for the production of biomass of microorganisms by incubation on a nutrient medium containing mineral salts and a carbon source in the form of liquid or solid waste from natural raw materials processing plants to produce high-quality, easily digestible and digestible protein-rich feeds and enriched with trace elements, vitamins, biologically useful substances, which have protective properties, with approx neniem microbial genera Lactobacillus and Propionibacterium fermentation culture medium with the accumulation of biomass, metabolism products, enzymatic systems that enrich food product.

The value of the microorganisms used by the method according to the invention is that they consume mono- and disaccharides, hydrolysis intermediates, pentosans and other organic substances. In the process of cultivation, they synthesize, in addition to biomass, vitamins, amino acids, complexes of enzyme systems with protective and prophylactic properties. The result is a protein feed enriched with biologically active components, as well as protective substances that provide long-term storage of feed in raw form.

The wet food obtained by the method according to the invention can be stored for 3-6 months without microbiological damage due to the presence of protective substances in it (organic acids, their salts, enzyme systems and complexes).

Their valuable property is the ability to develop on various complex environments, without requiring additional nutrients.

The microorganisms used are not spore-forming, therefore, neutralization of the culture fluid and wastewater is not required. According to SanPiN 2.3.2.1078-01 they are included in the list of substances that do not have a harmful effect on food products.

These microorganisms are anaerobes, therefore, air flow is not required, which makes the process highly economical.

By the method according to the invention, it is recommended to incubate pairs of cultures of lactic acid and propionic acid bacteria for 3-23.9 hours or for 24-50 hours. A shorter incubation time (3-23.9 hours) allows you to prepare food in the form of biomass of cells in phase exponential growth. When an animal enters the gastrointestinal tract, such living cells of lactic acid and propionic acid bacteria take root in it and are included in the regulation of metabolic processes, increase feed absorption and digestibility, and contribute to an increase in weight gain when feeding them to animals and birds, especially young animals. This food also contains protein, but in a smaller amount than food obtained by a longer incubation of microorganisms.

With the duration of the cultivation of microorganisms up to 24-50 hours, more vitamin B _{12 is} accumulated, which remains in the solid phase of the culture fluid, the volume of biomass increases, that is, the yield of feed enriched with protein.

The target product obtained as a result of the fermentation of microorganisms contains 35-50% dry matter protein, 30-40% amino acids, is enriched with vitamins E and group B, including B ₁₂ , enzyme systems and complexes that provide protective and preventive properties of the resulting feed.

If the process of growing biomass of lactic acid and propionic acid bacteria is carried out, for example, for 18 hours, the target product contains 35% s.v. protein, during fermentation 24 hours - 39%, for 48 hours - 45% s.v. During 18 hours of cultivation (exponential phase), protective substances, catalase-peroxidase complexes, accumulate in the culture fluid, therefore, the process takes up to 23.9 hours to obtain a feed product with protective properties, although protein accumulation in this case is less (35%).

Lactic acid and propionic acid bacteria in this case are in an active state of reproduction, which contributes to their favorable introduction from feed into the gastrointestinal tract of animals, and also have the greatest viability and stability when dried.

When bacteria are cultured for 24-50 hours, an intensive accumulation of biomass occurs with an increase in the protein content in the cells. The resulting product is more enriched with vitamins and amino acids.

The method according to the invention can be used to obtain high protein feed directly at the enterprises for the processing of natural raw materials or at the enterprises for feed production and can be used both in dried and in liquid form.

The fermentation process can be carried out both continuously and space-topping.

By the successive introduction of lactic acid bacteria and then propionic acid bacteria into the nutrient medium, the yield of the target product is increased due to the intensification of the growth of propionic acid bacteria on the medium, which contains the vital products of lactic acid bacteria.

The method is simple to execute. The nutrient medium is available, since it consists only of industrial wastes, to which only one cobalt salt is added, which reduces the cost and simplifies the method. The process does not require aeration, which reduces energy consumption. The method allows to utilize large volumes of industrial waste, turning them into a complete protein-vitamin feed.

The production of feed by the method according to the invention is waste-free.

It is possible to separate the culture fluid into a solid and liquid phase before drying by filtration through a filter press or vacuum filter and direct the solid phase with a humidity of (50-60)% to dry or use directly in the form of enriched feed, and use the liquid, for example, in alcohol production instead of water in a batch or in other stages of alcohol production.

The following are examples of the implementation of the method.

Example 1. In milk whey with a solids content of 4-5% add 0.9 mg / l CoCl ₂ , heated to 50 ° C, make 20% vol. cells of pure cultures of Lactobacillus acidophilus 1660/02 and Propionibacterium freudenreichii subsp. shermanii 103/12 in a ratio of 0.8: 1.2. Incubation is carried out for 3 hours at a temperature of 45 ° C and a pH of 5.9-6.0. After drying the obtained biomass, a protein-vitamin feed is obtained containing lactic acid and propionic acid bacteria cells in the exponential growth phase.

Example 2. The beer pellet is diluted with tap water in a ratio of 1: 1, the pH is set at 5.8-6.0, the temperature is 59-61 ° C and 1.4 u / g of conditional starch of α-amylase are added. The mixture is kept under these conditions for 55 minutes, and then for 55 minutes at a temperature of 74-76 ° C. After cooling the mixture to 59-61 ° C, 0.4 u / g of α-amylase are introduced into it and maintained under these conditions for 35 minutes. The mixture is cooled to 45-50 ° C and 5.5 u / g of conditional glucoamylase starch are added and kept at pH 5.0 until a reducing substance content of 6% is reached, after which 1.1 mg / L CoSO ₄ is added to the prepared beer fermentolizate , a mixture of pure cultures of Lactobacillus acidophilus 1660/02 and Propionibacterium acnes 1450/28, taken in a ratio of 1.2: 0.8 in an amount of 5% vol. Incubation is carried out with periodic stirring for 20 hours at a temperature of 50 ° C and a pH of 5.9-6.0.

The resulting biomass of lactic acid and propionic acid bacteria in the exponential growth phase is dried and protein-vitamin feed is obtained.

Cell viability in the finished dried protein feed was preserved.

The resulting protein-vitamin product contained protein 49.6% by d.v., amino acids 44.1% by d.v., carbohydrates 37.7% by d.v.

Example 3. By the method of example 2 receive high-quality protein feed. At the same time, 1.6 g / l of conditional starch of α-amylase is added to the aqueous suspension of beer pellets, the prepared mixture is kept for 65 minutes at a temperature of 59-61 ° C and a pH of 5.8-6.0, then 65 minutes at a temperature of 74-76 ° С, 0.6 u / g of conditional starch of α-amylase are additionally introduced and incubated for 40 min at 59-61 ° С, then 6.5 units / g of conditional starch of glucoamylase are introduced and the mixture is maintained for 60 min at 50 ° С and pH 5 , 5 to achieve a content of reducing substances of 10% to obtain a fermentolizate. 1.1 mg / l of CoSO _{4 is} added to the fermentolizate and a pure culture of Lactobacillus plantarum 578/25 is incubated for 1.5 hours on the prepared medium, and then a pure culture of Propionibacterium freudenreichii subsp is added to it. shermanii 103/12 and incubation continue for another 48.5 hours

Example 4. Milling waste is ground to a passage of at least 80% through a sieve with mesh diameter of 1 mm An aqueous suspension of crushed flour wastes is prepared by mixing them with water in a ratio of 1: 3.0. 20 u / g of dry matter of cellulolytic β-glucanase cellulolytic enzyme are added to the prepared suspension and the prepared mixture is kept at a temperature of (59-61) ° С and a pH of 6.0 for 55 minutes and then at a temperature of 105 ° С for 25 minutes to achieve a content of reducing substances of 3-5%, after which additionally add 8 u / g of β-glucanase cellulose and 5 u / g of conditional glucoamylase starch. The prepared mixture is maintained at a temperature of 45 ° C and pH 5.0 for 30 minutes until reaching a content of reducing substances of 7-10%.

After processing, the saccharified mass is used to incubate microorganisms Lactobacillus plantarum 578/25 and Propionibacterium freudenreichii subsp. shermanii 103/12, taken in the ratio of 1.2: 0.8 at a temperature of 45 ° C and a pH of 5.9-6.0. After 24 hours, a protein-vitamin product is formed with a mass fraction of protein up to 49.6% d.v., with an amino acid content of 44.1% d.v., a carbohydrate content of 37.7% d.v.

Example 5. According to the method of example 4, an aqueous suspension is prepared from ground milling flour and water taken in a ratio of 1: 3.5. 22 u / g of dry cellulose cellulolytic cellulase enzyme are added to the prepared suspension and the prepared mixture is kept at a temperature of 61 ° C and a pH of 6.5 for 65 minutes, and then at a temperature of 95 ° C for 35 minutes until the content of reducing substances is reached 3 -5%, after which an additional 10 u / g of cellulose cellulase and 7 u / g of conditional glucoamylase starch are added. The prepared mixture is maintained at a temperature of 50 ° C and a pH of 5.5 for 15 minutes until reaching a content of reducing substances of 7-10%. The mixture of cultures of microorganisms Lactobacillus plantarum 578/25 and Propionibacterium acnes 1450/28, taken in the ratio of 0.8: 1.2 and the fermentation process lasts 23.9 hours at a temperature of 37 ° C and a pH of 5.9-6.1.

Example 6. By the method of example 4, prepare an aqueous suspension of crushed flour wastes by mixing them with water in a ratio of 1: 3.5. To obtain the fermentolizate, 22 u / g dry cellulose cellulolytic enzyme xylanase are added to the prepared suspension and the prepared mixture is kept at a temperature of 61 ° C and a pH of 6.5 for 55 minutes, and then at a temperature of 105 ° C for 25 minutes until the content reaches reducing substances 3-5%, after which an additional 10 units / g of xylanase cellulose and 7 units / g of conditional glucoamylase starch are added. The prepared mixture is maintained at a temperature of 50 ° C and a pH of 5.5 for 30 minutes until reaching a content of reducing substances of 6-9%.

Strains of microorganisms are introduced into the nutrient medium sequentially, first Lactobacillus, and after 6 hours - Propionibacterium. The results indicate an increase in the biomass growth rate and an increase in yield (see table).

Change in biomass yield and cycle time during joint and sequential cultivation

The name of the strain The biomass yield,% Relative cycle time Pr. freudenreichii subsp.shermanii 100 1 Pr. freudenreichii subsp.shermanii Lact. plantarum (applied simultaneously) 130 0.84 Lact. Plantarum Pr. freudenreichii subsp.shermanii (introduced after 6 hours) 160 0.7

The table shows the biomass yield and duration of the process relative to similar indicators in the case of cultivation of one culture of propionic acid bacteria.

With the simultaneous introduction of two cultures, the process is characterized by an increase in the amount of biomass formed by 1.3 times, which indicates a greater accumulation of bacterial cells and a decrease in cycle time by 16% compared with the introduction of one culture.

With the successive introduction of cultures (first lactic acid, and then propionic acid), the amount of biomass formed increases by 1.6 times, and the cycle time is reduced by 30%.

Example 7. In the post-alcohol stillage, 1.1 mg / L CoCl _{2 is} added at a temperature of 45 ° C and seeded with pure cultures of the microorganisms Lactobacillus plantarum 578/25 and Propionibacterium acnes 1450/28, which are added to the nutrient medium in an amount of 20% by volume of the medium. Incubation is carried out with periodic stirring at a temperature of 40 ° C and a pH of 5.9-6.1 for 24 hours. The resulting target product contains a protein of 39% d.v., amino acids 28% d.v.

Example 8. Fruit squeezes with a humidity of 50-60% are diluted with water in a ratio of 1: 2, the mixture is heated to a temperature of 45-50 ° C, cellulase is added in an amount of 1.8 u / l and the resulting mixture is kept for 3 hours at a temperature of 50 ° C until reaching a content of reducing substances of 10-15% to obtain a fermentolizate. Add 0.9 mg / l CoCl ₂ and seeded with pure cultures of the microorganisms Lactobacillus plantarum 578/25 and Propionibacterium acnes 1450/28, which contribute to the nutrient medium in an amount of 20% by volume of the medium. Incubation is carried out with periodic stirring at a temperature of 40 ° C and a pH of 5.9-6.1 for 48 hours.

Example 9. Starch waste is treated similarly to grain raw materials and flour milling waste, excluding the grinding stage. When preparing the mixture - starch waste - water, the amount of water is regulated so as to obtain a suspension with a concentration of 14-16%. Get food rich in biologically active substances and protein.

Example 10. The culture fluid after fermentation using a consortium of microorganisms in example 7 is subjected to preliminary filtration and drying of the solid phase to a moisture content of 10-12%. After filtration, a precipitate with a moisture content of 50-60% is obtained, which can be used as a finished wet feed.

The dried product (enriched protein feed) has the following composition:

mass fraction of crude protein,% d.v. 45.9

mass fraction of carbohydrates,% d.v. 44,2

mass fraction of ash,% d.v. 2,5

total amino acid content,% 43.4

of them

lysine + histidine 2.15

arginine 1.25

aspartic acid 4.25

threonine 2.0

2.08 series

glutamic acid 14.84

methionine + cystine 1.3

glycine 2.6

proline 3.14

phenylalanine + tyrosine 2.65

Alanine 4.4

isoleucine + leucine 2.53

mass fraction of protein according to Barshteyn,% on d.v. 37.1

mass fraction of fat,% d.v. 5.1

total micronutrient content, mg / kg 21800

of them

phosphorus 5900

potassium 3500

sodium 500

calcium 11600

magnesium 1000

iron 750

cobalt 4.9

the content of vitamins, mg / kg

E (tocopherol) 43.5

B ₁ (thiamine) 3.3

B ₂ (riboflavin) 7.7

B ₃ (pantothenic acid) 35.3

B ₄ (choline) 700

B ₅ (nicotinic acid) 26.8

B ₆ (pyridoxine) 8.2

B ₉ (folic acid) 18.0

B ₁₂ (cyancobalamin) 34.4

exchange energy, MJ 13.4

feed units, kg 1.37

Example 11. Crushed grain, flour waste or starch waste is used to prepare fermentolysates by treating their aqueous suspensions with amylolytic enzymes. To do this, 1.4 u / g of conditional starch of α-amylase are added to the aqueous suspension, the mixture is incubated for 55 minutes at a temperature of 59 ° C and a pH of 5.8, then 35 minutes at 95 ° C, after which the mixture is cooled to 45 ° C. 0.4 u / g of conditional starch of α-amylase and 6.5 u / g of conditional starch of glucoamylase are introduced into it. The prepared mixture is kept for 30 minutes at a temperature of 45 ° C and pH 5.0 until a reducing substance content of 7% is obtained to obtain an enzymeolysate, on which pairs of cultures of lactic acid and propionic acid bacteria are incubated to obtain a complete protein-vitamin feed.

Example 12. According to the method of example 11, fermentolizates of crushed grain, milling waste and starch waste are prepared, but first 1.6 u / g of conditional α-amylase starch are added to the aqueous suspension, incubated for 65 minutes at a temperature of 61 ° C and a pH of 6.5, then 25 minutes at 105 ° C, after which 0.6 u / g of conditional starch of α-amylase and 5.5 u / g of conditional starch of glucoamylase are added. The prepared mixture is kept for 20 minutes. At a temperature of 50 ° C and a pH of 5.5 to achieve a content of reducing substances of 10% to obtain fermentolizates, on which pairs of cultures of lactic acid and propionic acid bacteria are incubated to obtain high-quality protein-vitamin feed.

Thus, the method according to the invention allows to obtain high-quality protein-vitamin feed containing live cells of lactic acid and propionic acid bacteria. The method is simple to implement, does not require increased energy consumption and provides complete utilization of a wide range of waste from industries for the processing of natural raw materials. Due to the high content of protective substances (organic acids), the feed can be stored raw for a long time.

Claims (13)

1. The method of producing protein-vitamin feed, which consists in the incubation of microorganisms in a nutrient medium containing mineral salts and a carbon source in the form of waste from the processing of natural raw materials or in the form of a fermentolizate waste to obtain the target product in the form of biomass of incubated microorganisms, while as of mineral salts use cobalt salt, grain, flour milling waste, starch waste, post-alcohol distillery distillery, beer pellet, fruit squeeze, are used as production waste whey, fermentolizate is prepared from grain, flour waste, starch waste, beer pellets or fruit marc, and incubated on a nutrient medium are cultures of lactic acid and propionic acid bacteria used in pairs: lactic acid and propionic acid bacteria combined or sequentially: first, lactic acid bacteria are incubated, and then continue to incubate lactic acid bacteria, and then continue joint incubation of lactic and propionic acid bacteria. 2. The method according to claim 1, characterized in that pairs of strains of lactic acid and propionic acid bacteria are incubated on a nutrient medium: Lactobacillus acidophilus 1660/02 and Propionibacterium freudenreichii subsp. shermanii 103/12; or Lactobacillus acidophilus 1660/02 and Propionibacterium acnes 1450/28; or Lactobacillus plantarum 578/25 and Propionibacterium freudenreichii subsp. shermanii 103/12; or Lactobacillus plantarum 578/25 and Propionibacterium acnes 1450/28. 3. The method according to any one of claims 1 to 2, characterized in that the pairs of lactic acid and propionic acid bacteria are introduced into the nutrient medium in the ratio (0.8-1.2) :( 0.8-1.2) in an amount of 5- 20 vol.%, And the incubation is carried out with periodic stirring at a temperature of 37-50 ° C and a pH of 5.9-6.0. 4. The method according to any one of claims 1 to 3, characterized in that during sequential incubation, the joint incubation of lactic acid and propionic acid bacteria is carried out after 1.5-6 hours of preliminary incubation of lactic acid bacteria. 5. The method according to any one of claims 1 to 4, characterized in that the target product with high biological value, enriched in living cells in the active growth phase, is obtained by incubating a pair of microorganisms in a nutrient medium for 3-23.9 hours, and the target product with a high protein content is obtained by incubation for 24-50 hours 6. The method according to any one of claims 1 to 5, characterized in that the fermentolizate of grain, flour waste or starch waste is obtained by treating it with hydrolytic enzymes: first cellulolytic, and then a mixture of cellulolytic and amylolytic, while the grain and flour milling waste are pre-crushed, the cellulolytic enzyme is introduced in stages: first, in an amount of 20-22 u / g dry matter of cellulose and the prepared mixture is kept at a temperature of 59-61 ° C and a pH of 6.0-6.5 for 55-65 minutes, then at a temperature of 95- 105 ° C for 25-35 min until the content of reducing substances reaches 3-5%, after which 8-10 u / g of dry matter of cellulose cellulolytic enzyme and 5-7 u / g of conditional starch of amylolytic enzyme are additionally added and the prepared mixture is kept at a temperature of 45-50 ° C and pH 5.0-5.5 for 15-30 minutes until the content of reducing substances reaches 7-10% to obtain the fermentolizate. 7. The method according to any one of claims 1 to 6, characterized in that the grain or milling waste is ground to pass at least 80% through a sieve with mesh diameter of 1 mm, and an aqueous suspension prepared by mixing the ground waste with water in enzymes is subjected to treatment ratio 1: (3.0-3.5). 8. The method according to any one of claims 1 to 5, characterized in that for their treatment with starch waste enzymes, a 14-16% aqueous suspension is prepared. 9. The method according to any one of claims 6 to 8, characterized in that β-glucanase, xylanase or cellulase is used as cellulolytic enzymes, and α-amylase, β-amylase or glucoamylase as amylolytic enzymes. 10. The method according to any one of claims 1 to 5, 7-9, characterized in that the fermentolizate of grain, flour waste or starch waste is obtained by treatment with amylolytic enzymes, while the waste is first treated with 1.4-1.6 u / g conditional starch of α-amylase for 55-65 minutes at a temperature of 59-61 ° C and a pH of 5.8-6.5, then for 25-35 minutes at a temperature of 95-105 ° C, after which 0, 4-0.6 u / g of conditional starch of α-amylase and 5.5-6.5 u / g of conditional starch of glucoamylase and incubated for 20-30 minutes at a temperature of 45-50 ° C and a pH of 5.0-5.5 to achievements content reduc ruyuschih substances 7-10% to obtain fermentative. 11. The method according to any one of claims 1 to 5, characterized in that to obtain the fermentolizate from the beer pellet, an aqueous suspension is prepared by dilution with water in a ratio of 1: 1 and treated with amylolytic enzymes: first, 1.4-1.6 units are introduced into it / g conditional starch - α-amylase and the prepared mixture is kept at a temperature of 59-61 ° C and a pH of 5.8-6.0 for 55-65 minutes, and then for 55-65 minutes at 74-76 ° C then 0.4-0.6 u / g of conditional starch of α-amylase are additionally introduced and the mixture is incubated for 35-40 minutes at 59-61 ° C, then 5.5-6.5 u / g of conditional starch of glucoamyl are introduced The mixture and the mixture are kept for 30-60 minutes at a temperature of 45-50 ° C and a pH of 5.0-5.5 until the content of reducing substances reaches 7-10% to obtain an fermentolizate. 12. The method according to any one of claims 1 to 5, characterized in that in order to obtain the fermentolizate from the fruit marc, they prepare an aqueous suspension in a ratio of 1: 2, 1.7-1.9 u / l of a pectolytic or cellulolytic enzyme are introduced into it and the resulting mixture was incubated for 2-3 hours at a temperature of 45-50 ° C until reaching a content of reducing substances of 10-15% to obtain the fermentolizate. 13. The method according to p. 12, characterized in that pectin esterase or polygalacturonase is used as pectolytic enzymes, and cellulase is used as the cellulolytic enzyme.