Method for preparing protein-vitamin fodder

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

 

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

The world shortage of protein to the beginning of the XXI century is estimated at 30-35 million tons per year. The main way of reducing this deficit is the production of biomass using microbial synthesis.

Grain raw rye, barley, wheat, milling waste, including substandard, used for animal feed or for the preparation of animal feed. However, the grain raw material is not a complete food because of the low content of protein, poor digestibility him, unfavorable ratio of proteins and carbohydrates. More promising is to obtain protein meal by growing the microorganisms in the grain raw materials.

A valuable raw material for the production of feed is waste industries, processing of natural raw materials, however, the rational utilization of waste virtually no. Partly waste sent for animal feed raw, but the duration of their storage not more than 1 day. Partially used drying, for example, alcohol stillage, spent grains, but this process is inefficient because of the large consumption of heat flow meters and low quality of the dried product (protein and other useful substances).

A method of obtaining protein and vitamin food by incubating the microorganism is in the nutrient medium, contains mineral salts and a carbon source in the form of waste production and processing of natural raw materials to produce the desired product in the form of biomass of microorganisms (Patent RU №2183666, CL 12 F 3/10, 12 R. 7/06, And 23 To 1/06, publ. 20.06.2002 g) [1].

As waste production and processing of natural raw materials in this known method uses a grain that is ground, subjected to enzymatic hydrolysis with getting fermentolizate, which are nutrient medium for growing yeast Saccaromyces cerevisiae with the purpose of obtaining alcohol. The resulting biomass is used in the feed.

However, the yeast is grown in aerobic conditions, which requires considerable energy to the air in the environment for incubation. For use of biomass in the feed it is necessary to subject it to a heat treatment with the goal of ending the life of the yeast. Such processing requires substantial resources and is accompanied by the destruction of part of the bioactive components of food. Such food contains no living cells, capable to settle in the gastrointestinal tract of the animal and to improve its performance.

A method of obtaining protein and vitamin food by incubation of microorganisms in a nutrient medium containing mineral salts and a carbon source in the form of waste production in the processing is the development of a natural raw material (Patent RU №2054881, class. And 23 To 1/165, publ. 27.02.96,) [2].

As waste production and processing of natural raw materials in this known case, use a ground-up grain and milling waste production, which is subjected to heat treatment, followed by culturing yeast of the genus Candida and bacteria, producing amylolytic enzymes, which carry out the enzymatic hydrolysis of milled raw material and waste milling.

The disadvantage of this known method is used for obtaining protein feed yeast of the genus Candida, which are conditionally pathogenic microorganism, which requires additional process of thermolysis and special equipment, as well as complex systems of neutralization of effluents and air emissions, requires a significant amount of air. Sanitary and epidemiological rules SanPiN 2.3.2.1078-01 yeasts of the genus Candida included in the list of substances that have harmful effects on human health.

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

A method of obtaining protein and vitamin food by incubation microorganis what s in a nutrient medium, containing distillery vinasse, obtaining biomass, which is mixed with the carbohydrate-containing component (USSR Author's certificate No. 1500671, CL 12 R. 7/06, publ. 1990) [3].

As microorganisms in this known case, use the yeast that requires aeration during incubation, and the food is subject to heat treatment, as it should not contain living yeast cells. This reduces the biological value of the feed.

A method of obtaining protein and vitamin food by incubation of microorganisms in a nutrient medium containing mineral salts and a carbon source in the form of waste production and processing of natural raw materials. (Patent RU №2159287, C 12 P 21/00, And 23 To 1/06, publ. 20.11.2001,) [4].

In this known method as waste use distillery vinasse, which make starch-containing raw material - waste grain-milling industry. The mixture is subjected to heat treatment in an acidic environment, while on the obtained hydrolysate grown biomass of bacteria Pseudomonas biforme BWA - 644.

However, the resulting biomass is subjected to heat treatment, which leads to reduction of its biological value. In the prepared stern no living cells, enriching the intestinal microflora of animals consuming the feed. Food is not subject to long-term storage is the in their raw form.

The technical result achieved by the present invention is the obtaining of 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, simplify the process, reduce energy consumption, improve the safety of raw feeding.

This technical result is achieved in that a method of obtaining protein and vitamin food is incubation of microorganisms in a nutrient medium containing mineral salts and a carbon source in the form of waste production and processing of natural raw materials or in the form of fermentolizate waste to obtain the desired product in the form of biomass inkubiruemykh microorganisms, as a mineral salt used is a salt of cobalt, as waste production using grain, milling waste, starch waste, distillery grains, brewer, fruit pomace or whey, fermentolizat made from grain, milling waste, starch waste, spent grains or fruit Marc, and incubated on a nutrient medium culture of lactic and propionic acid bacteria, used in pairs: together lactic and propionic acid bacteria or sequentially: first incubated m is lochmocola bacteria, and then continue co-incubation of lactic and propionic acid bacteria.

It is advisable to incubate in a nutrient medium a couple of strains of lactic 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.

A pair of lactic and propionic acid bacteria is recommended to bring in a nutrient medium in a ratio of(0,8-1,2):(0,8-1,2) in the amount of 5-20% by vol., and the incubation carried out at periodic stirring at a temperature 37-50°and a pH of 5.9 to 6.0.

Sequential incubation of co-incubation of lactic and propionic acid bacteria, it is advisable to carry out after 1.5-6 h pre-incubation of lactic acid bacteria.

Target product with high biological value, enriched by living cells that are in an active growth phase, can be obtained by incubation of a pair of microorganisms in a nutrient medium within 3-23,9 h, and the target product with a high content of protein - incubation during 24-50 hours

Fermentolizat grain, milling waste, or starch waste can be obtained by processing hydrolytic enzymes: first cellulolyticus, and then the mixture cellulolyticus and amylolytic is about, grain and milling waste pre-ground, cellulolyticus enzyme make step: first, in the amount of 20 to 22 u/g of dry matter of the pulp and incubated the mixture at a temperature 59-61°C and pH 6.0-6.5 for 55-65 min, and then at a temperature of 95-105°C for 25-35 minutes until a content of reducing substances 3-5%, then also add 8-10 u/g of dry matter of the pulp cellulolyticus enzyme and 5-7 u/g conditional starch amylolytic enzyme and the mixture maintained at a temperature of 45-50°C and pH 5.0-5.5 over 15-30 minutes before reaching the content of reducing substances 7-10% with obtaining fermentolizate.

Grain or flour waste is crushed to pass at least 80% through a sieve with a mesh diameter of 1 mm, and the processing enzymes are subjected to aqueous suspension prepared by mixing the waste with water in the ratio 1:(3,0-3,5).

For processing enzymes of starch waste prepare them 14-16%aqueous suspension.

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

Fermentolizat grain, milling waste, or starch waste can also be obtained by processing amylolyticus the mi enzymes, the waste is first treated with 1,4-1,6 u/g conditional starch α-amylase for 55-65 minutes at a temperature of 59-61°and a pH of 5.8 to 6.5, then for 25-35 minutes at a temperature of 95-105°C, after which further added to 0.4-0.6 u/g conditional starch α-amylase and 5,5-6,5 u/g conditional starch glucoamylase and incubated for 20-30 min at a temperature of 45-50°C and pH 5.0-5.5 to achieve content reducing substances 7-10% with obtaining fermentolizate.

To obtain fermentolizate from spent grains, it is advisable to prepare its aqueous suspension by dilution with water in the ratio 1:1 and process amylolytic enzymes: first enter into it is 1.4-1.6 u/g conditional starch α-amylase, and the mixture kept at a temperature 59-61°and a pH of 5.8-6.0 for 55-65 min, and then for 55-65 min at 74-76°C, after which an additional type of 0.4-0.6 u/g conditional starch α-amylase, and the mixture stand 35-40 min at 59-61°, then enter 5,5-6,5 u/g conditional starch glucoamylase and the mixture to stand for 30-60 minutes at a temperature of 45-50°C and pH 5.0-5.5 to achieve reducing substances 7-10% with obtaining fermentolizate.

To obtain fermentolizate of fruit pomace is recommended to cook them in water suspension in a ratio of 1:2, enter 1.7 to 1.9 u/l pectolytic or cellulolyticus enzyme and received MES to stand for 2-3 hours at a temperature of 45-50° To achieve the content of reducing substances 10-15% with obtaining fermentolizate.

As pectolytic enzymes can be used to pectinesterase or polygalacturonase, and as cellulolyticus enzyme is a cellulase.

The essence of the proposed method lies in the development of wasteless technology of production, providing for obtaining biomass of microorganisms by incubation in a nutrient medium containing mineral salts and a carbon source in the form of liquid or solid waste for processing natural raw materials to produce high-quality and easily digestible and assimilable food, rich in protein and enriched with trace elements, vitamins, biologically useful substances with protective properties, using microorganisms of the genera Lactobacillus and Propionibacterium for fermentation of a nutrient medium with the accumulation of biomass, products of metabolism, enzyme systems, enriching feed product.

The value of the microorganisms used in the method according to the invention, is that they consume mono - and disaccharides, intermediate products of hydrolysis of the pentosans and other organic substances. In cultivation they are synthesized, in addition to biomass, vitamins, amino acids, complexes of enzyme systems with protective and preventive properties. The result is protein feed, enriched with biologically active components, as well as protective substances, providing long-term storage of food in raw form.

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

Valuable feature is the ability to grow on a variety of complex environments, without requiring additional nutrient.

Used microorganisms are not spore-forming, therefore, neutralization of the culture fluid and the effluent is not required. According to SanPiN 2.3.2.1078-01, they are included in the list of substances that have no adverse impact on food products.

These microorganisms are anaerobic, so air flow is not required, making the process highly economical.

The method according to the invention it is recommended to incubate pairs cultures of lactic and propionic acid bacteria during 3-23,9 hours or during 24-50 hours shorter incubation period (3-23,9 h) allows you to cook food in the form of biomass of cells in the exponential phase of growth. When released into the gastrointestinal tract of an animal such living cells Molochnoe is small and propionic acid bacteria survive in it and included in the regulation of metabolic processes, increase feed intake and digestibility, increase of gain when fed to their animals and birds, especially the young. This food also contains protein, but in smaller numbers than the feed, get a longer incubation of microorganisms.

The duration of the cultivation of microorganisms to 24-50 hours accumulate more vitamin B12that remains in the solid phase of the culture fluid, increases the amount of biomass, i.e. the output of the feed, protein fortified.

The resulting fermentation of microorganisms of the target product contains protein 35-50% of dry matter, amino acids 30-40%, enriched with vitamins E and b group, including12enzymatic systems and complexes, providing protective and preventative properties of the obtained feed.

If the process of growing biomass lactic and propionic acid bacteria carried out, for example, for 18 h, the target product contains : protein 35% SV, fermentation time of 24 hours is 39%, 48 hours - 45% SV For 18 h of cultivation (exponential phase) accumulated in the culture fluid protective agent - catalase-peroxidase complexes, so to get a feed product with protective properties, the process is carried out to 23.9 hours, although the accumulation of the protein in this case is smaller (35%).

Molochnik the e and propionic acid bacteria in this case are active breeding that is favourable to the introduction of feed in the gastrointestinal tract of animals, but also have the highest viability and stability during drying.

The cultivation of bacteria during 24-50 hours of intensive biomass accumulation with 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 a high-protein feed directly on the plants for processing natural raw materials or on the enterprises production and be used both in dry and in liquid form.

The fermentation process can be both continuous and space-topping-up method.

Receiving sequential entry in a nutrient medium initially lactic acid bacteria, and then - propionic acid increases the yield of the desired product due to the intensification of the growth of propionic acid bacteria in the environment, which contains waste products of lactic acid bacteria.

The method is simple in execution. Nutrient medium is available, as it consists only of waste products to which you have added only one salt of cobalt, which reduces and simplifies the way. The process does not require aeration, which reduces energy costs. The method can utilize a large amount of the industrial waste, turning them into a complete protein and vitamin food.

Fodder production method according to the invention is free of waste.

Possible before drying the culture fluid to separate into solid and liquid phase by filtration through a filter press or vacuum filter and solid phase humidity (50-60)% to be directed to drying or to use directly in the form of enriched food and liquid to use, for example, in ethanol production instead of water for mixing or in other stages of alcohol production.

Below are examples of implementation of the method.

Example 1. In milk serum solids content of 4-5% add 0.9 mg/l CoCl2is heated to 50°To contribute 20% vol. cells of pure cultures of Lactobacillus acidophilus 1660/02 and Propionibacterium freudenreichii subsp. shermanii 103/12 in the ratio of 0.8 to 1.2. The incubation is conducted for 3 hours at a temperature of 45°and a pH of 5.9 to 6.0. After drying the obtained biomass get protein and vitamin food containing cells of lactic and propionic acid bacteria in the exponential growth phase.

Example 2. Brewer diluted with tap water at a ratio of 1:1, determine the pH between 5.8 to 6.0, temperature 59-61°and contribute to 1.4 u/g conditional starch α-amylase. The mixture is kept under these conditions for 55 min, and then 55 minutes at a temperature of 74-76°C. After cooling from the thou up to 59-61° With introducing her to 0.4 u/g α-amylase and maintained under these conditions for 35 minutes the Mixture is cooled to 45-50°and contribute to 5.5 units/g conditional starch glucoamylase incubated at pH 5.0 to achieve the content of reducing substances 6%, then cooked fermentolizat spent grains impose 1.1 mg/l CoSO4the mixture of pure cultures of Lactobacillus acidophilus 1660/02 and Propionibacterium acnes 1450/28 taken in the ratio of 1.2:0.8 to 5% vol. Incubation is carried out at periodic stirring for 20 hours at a temperature of 50°and a pH of 5.9 to 6.0.

The resulting biomass lactic and propionic acid bacteria in the exponential growth phase, dried and get protein and vitamin food.

Cell viability in the finished dried protein stern preserved.

The obtained protein-vitamin product contained protein 49.6% SV, amino acids to 44.1% for SV, carbohydrates 37,7% SV

Example 3. According to the method of example 2 get high quality protein food. In aqueous suspension of brewer's grains contribute to 1.6 g/l conventional starch α-amylase, and the mixture incubated for 65 min at a temperature 59-61°and a pH of 5.8-6.0, then 65 minutes at a temperature of 74-76°additionally impose 0.6 u/g conditional starch α-amylase and incubated for 40 min at 59-61°, then enter a 6.5 u/g conditional starch glucoamylase and mix well who live 60 min at 50° C and pH 5.5 to achieve the content of reducing substances 10% with obtaining fermentolizate. In fermentolizat contribute 1.1 mg/l SO4and the prepared environment 1.5 hours incubated pure culture of Lactobacillus plantarum 578/25, and then it adds a pure culture of Propionibacterium freudenreichii subsp. shermanii 103/12 and the incubation continued for another 48,5 h

Example 4. Milling waste is crushed to pass at least 80% through a sieve with a mesh diameter of 1 mm is Prepared aqueous suspension of powdered milling waste by mixing them with water in the ratio 1:3,0. In the prepared suspension is made of 20 u/g of dry matter of the pulp cellulolyticus enzyme β-glucanase and incubated the mixture at a temperature of (59-61)°C and pH 6.0 for 55 min, and then at a temperature of 105°C for 25 minutes to reach the content of reducing substances 3-5%, then also add to 8 IU/g cellulose β-glucanase and 5 u/g conditional starch glucoamylase. The mixture is maintained at a temperature of 45°C and pH 5.0 for 30 min before reaching the content of reducing substances 7-10%.

After processing osaharennoe mass used for incubation of the microorganisms Lactobacillus plantarum 578/25 and Propionibacterium freudenreichii subsp. shermanii 103/12, taken in the ratio of 1.2:0.8 a at a temperature of 45°and a pH of 5.9 to 6.0. After 24 hours, the formed protein-vitamin product mass is howling proportion of protein to 49.6% SV, with the amino acid content of 44.1% for SV, the carbohydrate content of 37.7% SV

Example 5. According to the method of example 4 is prepared aqueous suspension of powdered milling waste and water, taken in the ratio of 1:3.5. In the prepared suspension contribute 22 u/g of dry matter of the pulp cellulolyticus enzyme cellulase and incubated the mixture at a temperature of 61°C and pH 6.5 for 65 min, and then at a temperature of 95°C for 35 minutes until the content of reducing substances 3-5%, after which additionally contribute 10 IU/g cellulose cellulase and 7 u/g conditional starch glucoamylase. The mixture is maintained at a temperature of 50°C and pH 5.5 for 15 min before reaching the content of reducing substances 7-10%. In the mixture contribute to the culture of microorganisms Lactobacillus plantarum 578/25 and Propionibacterium acnes 1450/28 taken in the ratio of 0.8:1.2 and the fermentation process continues to 23.9 hours at a temperature of 37°and a pH of 5.9 to 6.1.

Example 6. According to the method of example 4 is prepared aqueous suspension of powdered milling waste by mixing them with water in the ratio of 1:3.5. To obtain fermentolizate in the prepared suspension contribute 22 u/g of dry matter of the pulp cellulolyticus enzyme xylanase and incubated the mixture at a temperature of 61°C and pH 6.5 for 55 min, and then at a temperature of 105°C for 25 min is t to achieve the content of reducing substances 3-5%, then also add to 10 u/g of pulp xylanase and 7 u/g conditional starch glucoamylase. The mixture is maintained at a temperature of 50°C and pH 5.5 for 30 min before reaching the content of reducing substances 6-9%.

Strains of microorganisms contribute to the nutrient medium sequentially, first, Lactobacillus, and after 6 hours - Propionibacterium. The results show an increase in the rate of growth of biomass and the increase in output (see table).

The change of biomass yield and cycle time in a joint and consistent cultivation

The name of the strainBiomass yield, %The relative cycle length
The WG. freudenreichii subsp.shermanii1001
Pr. freudenreichii subsp.shermanii Lact. plantarum (made simultaneously)1300,84
Lact. Plantarum Pr. freudenreichii subsp.shermanii (made in 6 hours)1600,7

The table below shows the yields of biomass and time consuming process as compared to the same indicators in the case of cultivation of a single crop propionic acid bacteria.

Upon simultaneous application of two cultures, the process is characterized by an increase in the number of the resulting biomass 1.3 the Aza, that proves greater accumulation of bacterial cells and the reduction of cycle time by 16% compared to making one culture.

The sequential introduction of crops (first, lactic acid, propionic acid and then) amount of produced biomass increased 1.6 times, and cycle time is reduced by 30%.

Example 7. In distillery vinasse add 1.1 mg/l l2at a temperature of 45°and inoculated With pure cultures of microorganisms Lactobacillus plantarum 578/25 and Propionibacterium acnes 1450/28 that contribute to the nutrient medium in the amount of 20% by volume environment. Incubation is carried out at periodic stirring at a temperature of 40°and a pH of 5.9 to 6.1 in the next 24 hours. The obtained target product contains protein 39% for SV, amino acids 28% SV

Example 8. Fruit pomace with a humidity of 50-60% is diluted with water in the ratio 1:2, the mixture is heated to a temperature of 45-50°With, in it add the cellulase in the amount of 1.8 u/l and the mixture was incubated for 3 h at a temperature of 50°before reaching the content of reducing substances 10-15% with obtaining fermentolizate. Add 0.9 mg/l CoCl2and seeded with pure cultures of microorganisms Lactobacillus plantarum 578/25 and Propionibacterium acnes 1450/28 that contribute to the nutrient medium in the amount of 20% by volume environment. Incubation is carried out at periodic paramasivan and at a temperature of 40° C and pH 5.9 to 6.1 in 48 hours.

Example 9. Starch waste is treated similarly raw grain and milling waste, excluding stage of grinding. When the preparation of a mixture of starch waste water, the amount of water is adjusted to obtain a slurry concentration of 14-16%. Get food, rich in biologically active substances and protein.

Example 10. Culture liquid after fermentation with the help of a consortium of microorganisms according to example 7 is subjected to pre-filtering and drying the solid phase to a moisture content of 10-12%. After filtration of the precipitate obtained by the humidity of 50-60%, which can be used as a ready wet food.

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

mass fraction of crude protein, % SV 45,9

mass fraction of carbohydrates, % SV 44,2

mass fraction of ash, % 2,5 SV

the total amino acid content, % 43,4

of them

lysine+histidine 2,15

arginine 1,25

aspartic acid 4,25

threonine 2,0

series 2,08

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 Burstein, % SV 37,1

mass fraction of fat, % SV 5,1

the total content of trace elements, mg/kg 21800

of them

phosphorus 5900

ka is s 3500

sodium 500

calcium 11600

magnesium 1000

iron 750

cobalt 4,9

the vitamin content, mg/kg

E (tocopherol) 43,5

B1(thiamine) 3,3

In2(Riboflavin) 7,7

In3(Pantothenic acid) 35,3

In4(choline) 700

In5(nicotinic acid) 26,8

In6(pyridoxine) 8,2

In9(folic acid) 18,0

In12(ciankobalamin) 34,4

the exchange energy, MJ 13,4

fodder units, kg 1,37

Example 11. The crushed grain, milling waste or starch waste is used to make fermentolizate by treating them with aqueous suspensions of amylolytic enzymes. To do this, in aqueous suspension contribute to 1.4 u/g conditional starch α-amylase, and the mixture is kept for 55 min at a temperature of 59°and a pH of 5.8, then 35 min at 95°C, after which the mixture is cooled to 45°With, give it to 0.4 u/g conditional starch α-amylase and 6.5 u/g conditional starch glucoamylase. Stand the mixture for 30 min at a temperature of 45°C and pH 5.0 to achieve the content of reducing substances 7% with obtaining fermentolizate, which is incubated for couples cultures of lactic and propionic acid bacteria with obtaining adequate protein and vitamin food.

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

Thus, the method according to the invention allows to obtain high-quality protein and vitamin food containing living cells of lactic and propionic acid bacteria. The method is simple to perform, does not require high energy consumption and ensures full utilization of a wide range of waste industries processing natural raw materials. Due to the high content of protective substances (organic acids) food can be stored long-term in its raw form.

1. The method of obtaining protein and vitamin food, which consists in the incubation of microorganisms in a nutrient medium containing mineral salts and a carbon source in the form of waste production and processing of natural raw materials or in the form of fermentolizate waste with obtaining the target product is in the form of biomass inkubiruemykh microorganisms, as a mineral salt used is a salt of cobalt, as waste production using grain, milling waste, starch waste, distillery grains, brewer, fruit pomace or whey, fermentolizat made from grain, milling waste, starch waste, spent grains or fruit pomace, are incubated in a nutrient medium culture of lactic and propionic acid bacteria, used in pairs: together lactic and propionic acid bacteria or sequentially: first incubated lactic acid bacteria, and then continue co-incubation of lactic and propionic acid bacteria.

2. The method according to claim 1, characterized in that incubated in a nutrient medium a couple of strains of lactic 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.

3. The method according to any one of claims 1 to 2, characterized in that a pair of lactic and propionic acid bacteria contribute to the nutrient medium in the ratio(0,8-1,2):(0,8-1,2) in the amount of 5-20 vol.%, and incubation is carried out at periodic stirring at a temperature 37-50°and a pH of 5.9 to 6.0.

4. The method according to any one of claims 1 to 3, characterized in that when the follower is the second incubation, co-incubation of lactic and propionic acid bacteria is carried out after 1.5 to 6 h pre-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 by living cells that are in an active growth phase, obtained by incubation of a pair of microorganisms in a nutrient medium within 3-23,9 h, and the target product with a high protein content get their incubation during 24-50 hours

6. The method according to any one of claims 1 to 5, characterized in that fermentolizat grain, milling waste, or starch waste produced by processing of hydrolytic enzymes: first cellulolyticus, and then the mixture cellulolyticus and amylolytic, while grain and milling waste pre-ground, cellulolyticus enzyme make step: first, in the amount of 20 to 22 u/g of dry matter of the pulp and incubated the mixture at a temperature 59-61°C and pH 6.0-6.5 for 55-65 min, and then at a temperature of 95-105°C for 25-35 minutes until a content reducing substances 3-5%, then also add 8-10 u/g of dry matter of the pulp cellulolyticus enzyme and 5-7 u/g conditional starch amylolytic enzyme and the mixture maintained at a temperature of 45-50°C and pH 5.0-5.5 over 15-30 minutes before reaching the content of reducing substances 7-10% with obtaining fermentolizate.

7. The method according to any one of claims 1 to 6, Otley is audica fact, what grain or flour waste is crushed to pass at least 80% through a sieve with a mesh diameter of 1 mm, and the processing enzymes are subjected to aqueous suspension prepared by mixing the waste with water in the ratio 1:(3,0-3,5).

8. The method according to any one of claims 1 to 5, characterized in that the processing enzymes of starch waste prepare them 14-16%aqueous suspension.

9. The method according to any of PP-8, characterized in that as cellulolyticus enzymes use β-glucanase, xylanase or cellulase, and as amylase - α-amylase, β-amylase or glucoamylase.

10. The method according to any one of claims 1 to 5, 7 to 9, characterized in that fermentolizat grain, milling waste, or starch waste produced by processing the amylolytic enzymes, the waste is first treated with 1,4-1,6 u/g conditional starch α-amylase for 55-65 minutes at a temperature of 59-61°and a pH of 5.8 to 6.5, then for 25-35 minutes at a temperature of 95-105°C, after which further added to 0.4-0.6 u/g conditional starch α-amylase and 5,5-6,5 u/g conditional starch glucoamylase and incubated for 20-30 min at a temperature of 45-50°C and pH 5.0-5.5 to achieve the content of reducing substances 7-10% with obtaining fermentolizate.

11. The method according to any one of claims 1 to 5, characterized in that to obtain fermentolizate of beer is Robina prepare its aqueous suspension by dilution with water in the ratio 1:1 and treated with amylolytic enzymes: first, give it a 1.4-to 1.6 u/g conventional starch - α-amylase, and the mixture maintained at a temperature 59-61°and a pH of 5.8-6.0 for 55-65 min, and then for 55-65 min at 74-76°C, after which further added to 0.4-0.6 u/g conditional starch α-amylase, and the mixture is kept for 35-40 min at 59-61°With, then give 5,5-6,5 u/g conditional starch glucoamylase and the mixture was incubated for 30-60 minutes at a temperature of 45-50°and pH 5,0-5,5 to achieve the content of reducing substances 7-10% with obtaining fermentolizate.

12. The method according to any one of claims 1 to 5, characterized in that to obtain fermentolizate of fruit pomace cook them in water suspension in a ratio of 1:2, it is injected 1.7 to 1.9 u/l pectolytic or cellulolyticus enzyme, and the mixture was incubated for 2-3 h at a temperature of 45-50°before reaching the content of reducing substances 10-15% with obtaining fermentolizate.

13. The method according to item 12, characterized in that the quality of pectolytic enzymes are used to pectinesterase or polygalacturonase, and as cellulolyticus enzyme is a cellulase.



 

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