RU1469613C - Method of bacterial preparation "propiovit" production - Google Patents

Abstract

The invention relates to a technology for the production of products of the microbiological industry and can be used in the food industry, veterinary medicine, and seven farm. The purpose of the invention is to accelerate the process and increase the yield of the target product. The bacterial preparation Propiovit based on the propionibacterium acnes VKPM - 1967 gropionic acid bacteria isolated from the rumen and their metabolic products was obtained by fermentation on a nutrient medium of the following coaav. wt.%: glucose 1.0 -1.5; corn extract 1.0 to 2.5; cobalt chloride 25 - 3.3 mg%; ammonium sulfate 0.05 - 25; tap water up to 100, for 40-48 hours at a temperature of 37 - 38 C and periodic stirring for 5-6 minutes to a maximum accumulation of biomass of propionic bacteria and the content of amine nitrogen 28-40 mg %, carbohydrates 0.56 - 0.8%, solids 2 - 5%. with a decrease in pH to 4.5-4D, after which the cupural liquid is averaged with a 40% NaOH solution, with a pH value of 6.5-7.0. mixed with protective additives and filler and dehydrated to a residual moisture content of not more than 5-8%. The living propionic acid bacteria and products of their vital activity, vitamins of group B and volatile fatty acids accumulated during fermentation, after application of protective additives and filler, are dehydrated to a residual moisture content of not more than 8%. This preparation is used by rpi for feeding animals and poultry to increase the balances of young farm animals and poultry, increase chickens, regulate stress after transport and regrouping, and in veterinary medicine as a preventive measure in the fight against dysbacteriosis of animals and poultry and Ketosis of cows. 1 3.U f 7-taba VO x C / 3 C

Description

The invention relates to a technology for producing products of the microbiological industry and can be used. but in the food industry, veterinary medicine and agriculture.

The aim of the invention is to speed up the process and increase the yield of the target product.,.

The invention is as follows.

To increase the nutritional value and quality of the bacterial preparation, a culture of Cicatricial propionic bacteria Proplonobacterium acnes, units of VKPM B-1967, and products of their metabolism, B vitamins and volatile fatty acids) grown on a modified glucose-corn medium of the following composition is used,%; glucose 1.0-1.5; corn extract 1.0-2.5; CoCl2 2.5-3.3 mg%; sulfuric acid ammonium 0.05-2.5; d.v. tap water dr 100. The pH of the nutrient medium before sterilization is set at 6.7-7.0, not after sterilization. below 6.3-7.0. Seed is applied in an amount of 7-10%. Cultivation is carried out for 40-48 hours at a temperature of 37-38 ° C with periodic stirring for 5-6 minutes. At the end of the growing period, the number of bacterial cages is 600–700 million / ml, the pH of the culture fluid decreases to 4.5–4.9, the content of amino nitrogen to 28.56–40.0 mg%, and carbohydrates up to 0.56-0.8%, solids up to 2-3%,

Cultural. a liquid with a pH of 4.5-4.9 is averaged with a 40% NaOH solution to a pH value of 6.5-6.8. Dehydration is carried out in a pseudo-boiling layer dryer at a temperature of 60-70 ° C after mixing with a protective medium and a filler or in a spray dryer (Anhydro) at a temperature of 130-140 ° C at the inlet after adding a protective medium and filler by concentrating the culture fluid at a solution temperature of 40-60 ° C to a dry matter content of 4-10% by mixing the concentrate with protective additives and filler and granulating the mixture, followed by drying of the wet granules in the fluidized bed at a temperature of 50-60 ° C to a residual moisture content of 5-8 %

Description of the process by stage m. 1. Growing. A culture of propionic acid bacteria is grown on a modified glucose-corn medium of the following composition,%: glucose 1.5; corn extract 1.0; Coda 3.3 mg%; ammonium sulfate 0.05; tap water up to 100 MP. PH value of the feed medium

before sterilization, set above 6.7-6.8, after sterilization - not lower than 6.3. In order to increase the viscosity of the nutrient medium and create more favorable conditions for the cultivation of propionic bacteria, 3-5% of excipients, for example flour, bran, malt sprouts, fodder yeast, as starch, flour, bran, etc. can be added to the nutrient medium Nitrogen compounds of malt sprouts and yeast are not only nutrients, but also serve to preserve bacteria during drying.

The nutrient medium is inoculated with a two-day culture. Seed is applied in an amount of 7-10%. The culture is grown in a fermenter for 48 hours at a temperature of 37 ° C with periodic stirring 8 for 5-6 minutes. By the end of the growing period, the number of bacterial cells is 600-700 million / ml, the pH of the culture fluid is reduced to 4.5-4.9; the content of amine nitrogen 28.56 mg%, carbohydrates 0.56%, solids 2%.

2. Concentration. Cultural. a liquid having a pH of 4.5–4.9 is averaged with a 40% NaOH solution to values of 6.5–6; 9, due to the fact that carbohydrates remain in the culture liquid after cultivation of propionic bacteria to 0.56 % and aluminum nitrogen up to 29.56 mg%, which can serve as protective substances, additional concentration of protective additives is not necessary during concentration.

Evaporation is carried out on a rotary-bag apparatus at a temperature of the evaporated solution of 40-50 ° С until the solids content in the concentrate is 4-10% (the volume of the initial culture liquid decreases by 2-5 times). Adjusting the pH of the culture fluid to neutral values is necessary to attenuate the inhibitory effect of the formed ..tjpopionic and acetic acids and to increase the yield of bacterial cells upon concentration of the culture fluid.

The carbohydrate content in the concentrate was 1.4%, and the amine nitrogen content was 46.73 mg%: In the course of concentration, the dependence of the survival of propionic bacteria on temperature and evaporation rate was studied. Before concentration, the pH of the culture fluid was adjusted to 6.3-7.0 (Table 2) after 48 hours of cultivation.

3. Granulation of the mixture of the concentrate and filler wire i with l; icitium medium, and without POR.

When granulating without a protective environment, bacterial survival reaches 70.5-94%. The addition of only 1% molasses gives a 100% survival rate in the granulation process.

The effect of traditional protective additives during granulation gave a positive effect, but in this case it can be limited to molasses only, without using milk powder, etc.

A. The effect of the protective medium is especially pronounced when drying wet granules. If during drying the survival rate was 19.85-28.2%, then the addition of 1% molasses increased the number of viable cells by 2.3 times (Table 4).

Important points when drying wet granules are the temperature of the coolant and the drying time. The wet granular product (with a humidity of at least 30%) is dried in a fluidized bed at a temperature of 40-60 C for 4-8 minutes. The resulting dry product in the form of small granules of light yellow color should have a residual moisture content of not higher than 8%.

Table 5 shows the average results over the entire process chain to obtain the final product.

Studies have shown that it is advisable to carry out the drying process of the granular product with this method at an air temperature of 50-60 ° C for 6-8 minutes. The resulting product should have a moisture content of not more than 8%.

PRI me R 1. Two-day fresh culture of Prop.acnes propionic acid bacteria with a number of cells of 450 million / ml are seeded in a fermenter with a nutrient medium of the following composition, wt.%: Glucose 1,5; corn crustact 1.0; ammonium sulfate 0.05; CoCl2 3.3 mg%; tap water, the pH of the nutrient medium before sterilization is not higher than 7.0, after sterilization not lower than 6.3. Cultivation was carried out at a temperature of 48 hours with periodic stirring every 12 hours for 6 minutes. By the end of the cultivation, the culture fluid contains at least 900 million / ml propionic bacteria cells; 0.56 carbohydrates; 28.56 mg% amine nitrogen; up to 2% solids; The pH of the culture fluid is reduced to 4.5-5.0.

Before concentration, the pH of the culture fluid was adjusted to 6.5 with a 40% NaOH solution. The culture fluid was evaporated 2 times at a temperature of 40 ° C to a solids content of A-5%. Carbohydrate Concentrate

1.4%, and amine nitrogen 46.73 mg%. The number of cells in 1 ml of concentrate was 1-1.2 billion.

A protective medium (molasses) is added to the volume of the concentrate (at the rate of 1%) 5 and corn flour in the ratio of 3; 7 as a filler. Humidity of the crude product is 30%.

Wet granular preparation

10 dried at a temperature of 50-60 ° C for

6-8 min to a residual moisture content of not more

b / o / b.

A dry granular product of light yellow color with a moisture content of 8% and a content of propionic bacteria in 1 g of at least 1 mld. Packed in plastic bags of 10 kg.

PRI me R 2. The grown culture of propionic bacteria, as in example 1, the number of cells in 1 ml of 500 million, concentrated to a solids content of 10%, granulated and dried under the same conditions as in example 1.

PRI me R 3. The culture fluid

5 propionic acid bacteria obtained by the described method in example 1 are neutralized to pH 6, €, mixed with a protective medium and sprayed onto a filler (cornmeal, bran, etc.) followed by drying in a pseudo-boiling layer at 60 -70 ° C. The culture fluid and the excipient are mixed in a ratio of 1; 1. To increase the yield of viable cells of propionic bacteria, you can use

5 combined filler, for example,

in the following composition; milk powder corn flour - fodder yeast 1; 1; 1.

PRI me R 4. The culture fluid.

obtained as in example 1, neutralize

40% NaOH solution to pH 6.8 t, dried by adding 3% molasses as a protective medium and filler, for example cornmeal, fodder yeast, etc. on a spray dryer (Anhudro) at air temperature at the inlet 130-140 ° С and 70-75 ° С at the outlet. The result is a dusty product with a moisture content of not more than 8.0% and a bacterial count of 1.0 billion / g.

0 when the dispersion of the spray affects the quality of the dry (product found that. Disk rotation speed of 46 thousand rpm gives better performance than 26 thousand rpm.

5 It is intended to use a preparation obtained by the proposed method and containing both living cells of propionic bacteria and their metabolic products (B vitamins and volatile fatty acids) in animal husbandry and poultry farming

to increase the body weight of young farm animals and poultry, and to increase chickens. regulation of stress after transport and regrouping, and in, veterinary medicine as a prophylactic in the fight against dysbacteriosis of animals and birds and ketosis of cows.

(56) USSR Copyright Certificate No. 1236630. class A 61 K 35/66, 1985.

The effect of fillers in a nutrient medium on the growth of propionic bacteria during cultivation

The effect of medium acidity on the survival of propionic acid bacteria cells during concentration

Table 1

table 2

Preparation of propiovit in granular form

Table 3

Table 4

Table 5

The survival rate of propionic bacteria upon drying in a pseudo-boiling layer with different fillers

Passed Eat 1 R Gabpitz S

Table6

Table 7 The survival rate of propionic bacteria by spray drying

claims a

Claims (4)

1. A METHOD FOR PRODUCING A PROPIOVIT BACTERIAL PREPARATION, which involves the cultivation of the Proplonlbacterlum acnes VKPM B-1967 strain on a nutrient medium containing glucose, corn extract, cobalt chloride, ammonium sulfate and water, at a temperature of 37 to 38 C to the maximum accumulation of nitrogen biomass mixing with protective additives and drying. Characterized in that, in order to accelerate the process and increase the yield of the target product, the cultivation is carried out on a medium containing these components in the following ratio, wt.%: Glucose 1.0-1.5; corn extract 1.0 to 2.5; cobalt chloride 2.5–3.3 mg%, ammonium sulphate 0.05–2.5 and tap water up to 100, until amine nitrogen reaches 29–40 mg%, carbohydrates 0.56–0.8% and solids 2 -5%, pH 5 of the culture fluid was adjusted with a 40% sodium hydroxide solution to 6.5-7.0, dehydration was carried out to a residual moisture content of 5-8%. 2. The method according to claim 1, characterized in that dehydration is carried out in a pseudo-boiling layer at a temperature of 60 - 70 C. 3. The method according to claim 1, characterized in that the dehydration is sprayed , method at a temperature of 130 - 140 C at the inlet. 4. The method according to claim 1, characterized in that dehydration is carried out by concentrating the culture fluid at a solution temperature of 40-50 ° C to a dry matter content of 4-10%, mixing the concentrate with protective additives and filler, and granulating the mixture, followed by drying of the wet granules in the thickest layer at a temperature of 50 ° C at the inlet to a residual moisture content of 5-8%.