SU1416100A1 - Method of cleaning biomass of microorganisms - Google Patents

Abstract

The invention relates to microbiology, in particular to methods for processing biomass of microorganisms and for obtaining protein products (BP) for food purposes, and can be used in the food, chemical and medical industries. The aim of the invention is to improve the quality of the protein product while obtaining lipid (LC) and nucleotide (NC) components of the biomass. The invention includes a two-stage treatment of biomass with aqueous solutions of lower alcohols with their content in the liquid phase of the system 50-75% in the first stage and 80-96% in the second stage. The alcoholic extracts are separated, combined, and after vacuum evaporation of the solvent, a drug LK or microbial fat containing at least 75% lipids is obtained. Microbial fat containing 85-90% lipids can be isolated from the alcohol extract obtained in the second stage of alcohol extraction. Fat-free material is treated with aqueous solutions of acids at a pH of 0.7-1.5 for 1-5 minutes at 50-100 ° C and neutralized to a pH of 2.5-4.5. The acidic extract is separated, dried and a NK preparation containing about 70% of nucleotides is obtained. With a high consumption of acid and alkali, the acidic extract is free of low molecular weight components by ultrafiltration or gel filtration before drying. Cell masses are washed with water, dried and get BP with a protein content of 65-90%, lipids and nucleic acids no more than 1% and with a high biological value. 1 tab. i (/) 4i CD

Description

This invention relates to microbiology and biotechnology, namely

I

methods of processing the biomass of micro-I organisms and the production of lipid and nucleotide proteins from it, and can be | used in the food, chemical, and medical sectors.

The purpose of the invention is to improve the quality of the protein product while simultaneously obtaining lipid and nucleotide components.

The method consists in the following: alcohol extraction of biomass is carried out in two stages with the content of alcohol in the liquid phase in the first stage 150-75%, and in the second stage 80-96%,; alcohol extracts are combined n you; of these, lipid components, i-fat protein material, are made with aqueous solutions of acids; at a pH of 0.7-1.5 for 1-5 min at; 50-100 ° C, neutralized to a pH of 2.3-4.5 J and the nucleotide components are isolated from the extract obtained.

The method is carried out as follows.

 The biomass of microorganisms, such as yeast, bacteria, fungal mycelium grown on alcohols, n-para-ny, carbohydrate-containing substrates, is subjected to a two-stage extraction with a lower alcohol, such as methanol, ethanol, propanol, preferably ethanol.

 At the first stage, the biomass is treated in a ratio of 1: 3 - 1:15 with anhydrous alcohol and water-alcohol solution of such a concentration so that the alcohol content in the liquid phase of the system is 50-75%. .

The processing temperature is maintained at 20-C, preferably 40-60 C, and the treatment time is 30-60 minutes At the end of the treatment, the extract is separated by a known method.

At the first stage of alcohol treatment, predominantly low molecular weight hydrophobic components, including toxic hydrophobic peptides poorly extracted by concentrated alcohols, are transferred to the solvent. This achieves the following results.

Biological degradation occurs

The value of the protein product. This is due to the fact that hydrophobic peptides

0 5

ABOUT

0 5

five

0

The dyes are enriched with melocene and food-grade amino acids and do not contain cysteine and methionine. As a result, the protein product is enriched with limiting amino acids. The amino acid isomers are not formed.

In addition, this treatment provides cleaner nucleotide components, since some of the hydrophobic components dissolve at high pH and can be extracted during subsequent acid treatment.

In the second stage, alcohol extraction of the biomass in a ratio of 1: 3-1:15 is treated with anhydrous alcohol or a water-alcohol solution of such a concentration so that the alcohol content in the liquid phase of the system is

80-96%. one

The processing temperature is maintained at 20–60 ° C; the processing time is 30–60 min.

The advantage of the second stage of alcohol extraction, besides removing lipids, is that it helps to increase the purity of the product of nucleotide components by reducing the extractability of protein substances. In this treatment, more than 90% of the lipids contained in the biomass are transferred to the solvent.

The suspension is separated in a known manner, for example, by centrifugation, filtration, sedimentation, into liquid and solid phases and an alcoholic extract of lipids and a defatted product are obtained.

The alcoholic extracts obtained in the first and second stages are combined, the solvent is distilled off under vacuum, and commercial microbial fat products containing up to 75% lipids are obtained.

To obtain a denuclearized protein product and a preparation of nucleic components, the defatted and dried .dl to remove residual solvent the cellular product is mixed in a ratio of 1: 5-1: 20, preferably 1: 5-1: 10, with an aqueous acid solution of such a concentration that the system has a pH of 0.7-1.5. The system is heated to 50-100 ° C, held for 1-5 minutes, neutralized, for example, with alkalis to a pH of 2.5-4.5 and in a known manner, for example, with a 31A161004 centrifuge

girovaniya, filtration, sedimenta-. Example 1 (in a known manner, divided into protein product). To 100 g of spray dried and nucleic acid extract. the biomass of bacteria Methylobacillus

When treating yeast and fungal methylophilus, the strain BWA-792 with a moisture content of biomass is preferable, more severe than 3%, with a protein content of 67% abs. These conditions, bacterial biomass of r.v., nucleic acids of 10% abs.s.c., are processed in softer uelipids 10% abs.c.., carbohydrates VIAH.4% abs.s.. 300 g of methanol are poured

As acids, preferably 10 g and, with stirring, 10 g of gaseous hydrochloric acid, edible ammonia, are introduced. The alcohol content of orthophosphoric acid thermal pi in the liquid phase of the system is 99%, and the alkaline solution, and the neutralization is carried out by the hydro- ratio of the solid phase: liquid sodium fluoride. 1: 3: 1. The obtained acid-alkaline treatment suspension is kept at 20 ° C for 30 minutes for several tasks. The cell product is separated from alcohol and improves the extraction of the extract by filtration. The semi-biological value of the protein protein is 249 g of the paste with a moisture content of 35%, duct due to the fact that it increases the resuspended content of cysteine and methionine, 20 nol, in 300 g. The alcohol content in the liquid phase isomers is not formed. Of the system, it is 99.8%, secondly, it allows you to extract more solid phase: the liquid phase is 90% of nucleic acids, 1: 5.3.

in the cell masses, and the alcohol extract is separated from the decrease in protein extract. 25 cell product by filtration and

Thirdly, an additional combination with the alcohol-ammonia extraction of the protein product from the litter takes place. The combined extract of the free components is obtained from the solvent by vacuum extraction of the nucleotide components of the parks. The solvent used is spray dried or freeze-dried. After vacuum evaporation by po I I chu but they also receive a marketable product containing 17 g of the vat residue, containing 70% of unsaturated sugar content. nucleic 35% lipids, 26% protein, 17% nucleic acids, new acids.

If at the stage of acid treatment, the residue of the extracted kdeki to create the required pH, the mass of the mass is dried from the solvent, a significant amount of acid is passed under vacuum and 82 g of degreased are obtained, for example, in case of using a cellular product, organic acids, and neutral - For nucleic acid extraction, lysing is carried out with weak bases, the cellular product is mixed with water, the acid extracts contain a large ratio of 1:10 and an intense amount of salts. In this case, it is stirred. In the injection system, the extracts are purified from a pH of 6.9.

co-molecular components, for example, the suspension is heated in the reactor before gel filtration or ultra- .50 ° C, held for 5 minutes, cooled by filtration. The desalted product is 45 ° separated solid phase and holds up to 75% abs. at. nucleotide extract by centrifugation, components. 635 ml of aqueous extract are obtained

 The protein product is washed with water with a dry matter content of 3%, in and dried by sublimation or spray including b.elk 43% abs.v., nucleotide. The yield of the product reaches other inoic acids 31% abs.v., lipids 70% of the original biomass, content 1% abs.v., carbohydrates 6% abs.s.. Protein ranges from 65-70% for yeast and fungal biomass and up to 90%. The pasteobacid cell mass is pro bacterial, the contents are lipidated with water and the sublimation is dried to 0.5-1.0% abs. in., - onno. 65 g of protein-derived nucleic acids are obtained — 0.3–0.7% abs.s.w. with a moisture content of 5%, protein content. The biological value of the protein content is 85% abs.s., the product’s nucleic acids exceed the biological one. , 5% abs.s.v., lipids 4% abs.s.v., the value of the original biomass. Carbohydrates 4% abs.s..

The table shows the biological value, protein content (B), Nucleic acids (N / K) and lipids (L) (L) in the original biomass, protein g product, vat residue and water. extract.

: From the data in the table it can be seen that during the processing of biomass under the described conditions, the protein content in the protein product increases by 18% (from 67 to 85%) as compared with the initial biomass; % (from 75% to 55%). Besides,

ki receive 14 g of the cubic residue representing microbial fat with a lipid content of 75%, protein 10%, nucleic acids 4%.

The remainder of the extracted glue-. exact masses are dried from the solvent under vacuum and get 81 g of fat-free product.

Fat-free material is mixed with 810 ml of an aqueous solution of phosphoric acid with a concentration of 0.1 g-mol / L. The system is adjusted to pH. Further processing of the suspension, the fast food product contains 4% lipids, 15 d t in two sequentially connected - iiTo significantly exceeds the norms of IP tubular heat exchangers. AT

first, the suspension is heated to 70 ° C and held for

min

Ol

secondly, cooling to 25 | iMH USSR.

j The bottom residue after vacuum-extraction of methanol contains 26% of protein,

(17% nucleic acids and 35% lipids, 20, After cooling, the suspension | 1 that cannot be isolated is neutralized with a 20% aqueous solution of the Preparation of lipid components. Rum sodium hydroxide to pH 3.5 and

separating the solid phase and the extract by centrifugation.

Acid extract. Ultrafiltrate onto a membrane with a molecular weight of 10,000 dal.

1 Aqueous extract contains 43% of protein and only 31% of nucleotide components that cannot be separated.

s in the form of the drug nucleotide components.

Example 2. In the first stage of alcohol extraction, 380 g of pressed Saccharomyces cerevisiae yeast strain 14-L-1 with a humidity of 75%, a protein content of 44% abs.s., nucleic acids 10% abs.s., lipids 12% abs.s.v., carbohydrates 14% abs.s.v. 1040 g of ethanol (rectified) and 100 g of water are poured. The alcohol content in the liquid phase of the system is 70%. The ratio of the solid phase: the liquid phase of 1:15. The resulting suspension is stirred with continuous stirring for 60 minutes and the cell product is separated from the alcoholic extract by filtration. Get 297 grams of pasta with a content of 30% abs.s.v.

In the second stage of alcohol extraction, 1117 g of ethanol (rectified) and 10 g of water are added to the paste. The alcohol content in the liquid phase of the system is 92%, the solid phase: liquid phase ratio is 1:15. The resulting suspension is maintained at 60 ° C for 60 minutes and the cell product is separated from the alcoholic extract by filtration.

Alcohol extracts obtained

in the first and second stages, f -

and free of solvent by vacuum evaporation. The solvent is reused. After a vacuum vyparton. Retettat is washed with water and dispensed sublimatically. Get 12 g

thirty

40

product with a content of nucleotide components of 75% abs. century, protein 12 12% abs. century, lipids 8% abs. century. Sponge-like cell mass is washed with water and the diluent is dried-2g but. Get 60 g of the protein product with a moisture content of 5%, a protein content of 70% abs.s., nucleic acids 0.8% abs.s., lipids 0.9% abs.s., carbohydrates 20% abs .with.

From the data in the table it can be seen that, when processing yeast biomass under the described conditions, the protein content in the protein product increases by 26% (from 44 to 70%) compared to the initial biomass, while by a known method this indicator increases only by 18%. the value increases by 10% (from 75 to 85%), while by a known method this indicator decreases by 20%. The content of lipids and nucleic acids does not exceed 1% and satisfies the requirements of IE AMN SSSR and FAO (WHO).

Microbial zhi.r contains 75% lipids (compared to 35% in the bottom 45

50

55

produced by a known method) and is a product suitable for technical use without additional.

ki receive 14 g of the cubic residue representing microbial fat with a lipid content of 75%, protein 10%, nucleic acids 4%.

The remainder of the extracted glue-. exact masses are dried from the solvent under vacuum and get 81 g of fat-free product.

Fat-free material is mixed with 810 ml of an aqueous solution of phosphoric acid with a concentration of 0.1 g-mol / L. The system is adjusted to pH. Further processing of the suspension

Ol

second, cooling to. After cooling, the suspension is neutralized with a 20% aqueous solution of sodium hydroxide to a pH of 3.5 and

tone. Retettat is washed with water and dispensed sublimatically. Get 12 g

0

0

a product with a content of nucleotide components of 75% abs.s.., protein 12 12% abs.s., lipids 8% abs.v. Sponge cell mass is washed with water and dried quickly. Get 60 g of the protein product with a moisture content of 5%, a protein content of 70% abs.s., nucleic acids 0.8% abs.s., lipids 0.9% abs.s., carbohydrates 20% abs .with.

From the data in the table it can be seen that, when processing yeast biomass under the described conditions, the protein content in the protein product increases by 26% (from 44 to 70%) compared to the initial biomass, while by a known method this indicator increases only by 18%. the value increases by 10% (from 75 to 85%), while by a known method this indicator decreases by 20%. The content of lipids and nucleic acids does not exceed 1% and satisfies the requirements of IE AMN SSSR and FAO (WHO).

Microbial zhi.r contains 75% lipids (compared to 35% in the bottom 5

0

five

produced by a known method) and is a product suitable for technical use without additional.

. The preparation of nucleotide components contains 75% of nucleotides (compared with 31% in the aqueous extract obtained by a known method) and is an intermediate product suitable for use in chemical industry.

Example 3. In the first stage of alcohol extraction to 100 g of spray-dried biomass of bacteria Methylobacillus methylophilus strain BWA-792 with a moisture content of 3%, a protein content of 67% abs.v., nucleic acids 10% abs.v., lipids 10 % abs.v. carbohydrates 4% abs.s.v. 162 g of isopropanol (azeotrol of 88% concentration) and 118 g of water are poured. The alcohol content in the liquid phase of the system is 50%; the solid phase ratio is:

liquid phase 1: 3. The obtained suspension is “a comparison of the achieved results

kept at 20 ° C for 30 minutes with continuous stirring and the cell product is separated from the alcoholic extract by separation. Get 255 g of pasta with a content of 35% abs.s.v.

In the second stage of alcohol extraction, 719 g of isopropanol (azeotrope 88% concentration) and 10 g of water are added to the paste. The alcohol content in the liquid phase of the system is 80%, the solid phase: liquid phase ratio is 1:10. The resulting suspension is held at 20 ° C for 30 minutes with continuous stirring and the cellular product is separated from the alcoholic extract by separation.

The extract obtained in the second stage of alcohol extraction is freed from the solvent by vacuum evaporation. 10 g of the cubic residue are obtained, representing microbial fat with a lipid content of 80%, protein of 10%, nucleic acids of 2%. .

The residue of the extracted cell masses is dried from the solvent under vacuum, and 78 g of defatted product are obtained.

The fat-free material is mixed with 390 ml of an aqueous solution of edible hydrochloric acid with a concentration of 0.07 g-mol / l. The system has a pH of 1.5. Further processing of the suspension is carried out in two series-connected tubular heat exchangers. In the first, the system is heated to 50 ° C and aged for 5 minutes in the second - cooling, to 25-30 ° C. After cooling, the suspension is neutralized with an aqueous 20% solution

thirty

can be carried out not only relative, but also in absolute terms.

From the data in the table it can be seen that during the processing of bacterial biomass, in the following conditions: the protein content in the protein product increases by 23% (from 67 to 90%) compared to the initial biomass, and by 5% (compared to the known method) to 90%), the biological value increases, respectively, by 15% (from 75 to 90%) and by 35% (from 55 to 90%). The content of lipids and nucleic acids does not exceed 1% and satisfies the requirements of 35 m of IP AMS SSSR and FAO. (WHO).

Microbial fat contains 80% lipi-dv (compared to 35% in the bottoms obtained by a known method) and is a product suitable for technical use without further purification.

The preparation of nucleotide components contains 75% of nucleotides (compared to 31% in the aqueous extract obtained by a known method) and is an intermediate product suitable for use in chemical industry.

40

45

50

55

Example 4. In the first stage of alcohol extraction, 100 g of sublimation-dried biomass of Fusarium Penicillium fungi with a moisture content of 2%, a protein content of 38% abs., Nucleic acids 8% abs. In., Lipids 10% abs. . C. in., carbohydrates 30% abs.v. 735 g of methanol and 243 g of water are poured. The alcohol content in the liquid phase of the system is 75%,

sodium hydroxide to pH 4.5 and separate the solid and the extract by centrifugation.

Acid extract is spray dried and get 11 g of the product with the content of nucleic acids 75% abs.s.v., protein 9% abs.v., lipids 9 abs.v.

The pasty cell mass is washed with water and dried sublimated. Get 70 g of the protein product with a moisture content of 5%, a protein content of 90 abs.v.v., nucleic acids

0.7% abs.s.v., lipids 0.5% abs.s.v., carbohydrates 1% abs.s.v.

In example 3, the same bacterial biomass was used. as in example 1 by a known method, then comparing the achieved results

can be carried out not only relative, but also in absolute terms.

From the data in the table it can be seen that during the processing of bacterial biomass, in the following conditions: the protein content in the protein product increases by 23% (from 67 to 90%) compared to the initial biomass, and by 5% (compared to the known method) to 90%), the biological value increases, respectively, by 15% (from 75 to 90%) and by 35% (from 55 to 90%). The content of lipids and nucleic acids does not exceed 1% and satisfies the requirements of IE AMS SSSR and FAO. (WHO).

Microbial fat contains 80% lipi-dv (compared to 35% in a bottoms obtained by a known method) and is a product suitable for technical use without further purification.

The preparation of nucleotide components contains 75% of nucleotides (compared to 31% in the aqueous extract obtained by a known method) and is an intermediate product suitable for use in chemical industry.

Example 4. In the first stage of alcohol extraction, 100 g of sublimation-dried biomass of Fusarium Penicillium fungi with a moisture content of 2%, a protein content of 38% abs., Nucleic acids 8% abs. In., Lipids 10% abs. . C. in., carbohydrates 30% abs.v. 735 g of methanol and 243 g of water are poured. The alcohol content in the liquid phase of the system is 75%,

solid phase: liquid phase ratio 1:10.

; The resulting suspension is kept at 50 ° C for 40 minutes and the cellular product is separated from the alcoholic extract by separation. Get 267 grams of rastay with a content of 35% abs.s.v ..; In the second stage of alcohol extraction, 947 g of methanol and 1.5 g of water are added to the paste. The alcohol content | In the liquid phase of the system is 95%. The ratio of the solid phase; liquid | phase 1:12. The resulting suspension is extracted (kept at 60 ° C for 30 minutes and the cell product is separated from the alcohol of the jTOBoro extract by separation. I The alcoholic extracts obtained in the first and second stages are combined and freed from the solvent | by vacuum evaporation The solvent is reused. After vacuum-I evaporation, 11 g of the bottom oil is obtained, representing microbial fat with a lipid content of 75%, protein 6%, 6% nucleic acids, I Residue of the extracted cellular masses is dried from the solvent under vacuum, get 87 g of degreased KTA

; The fat-free material is mixed with 1740 ml of an aqueous solution of phosphoric acid thermal food with a concentration of 1.0 g-mol / l. The system has a pH of 0.7. Further processing of the suspension is carried out in two series-connected tubular heat exchangers with an excess pressure of 1 MPa. In the first, the system is heated to

and incubation for 1 min, in the second - cooling to 25-30 ° C. After cooling, the suspension is neutralized with a 20% aqueous solution of sodium hydroxide to a pH of 2.5 and the solid phase and extract are separated by centrifugation.

Acid extract is subjected to gel filtration on Sephadex G-25. The nucleotide components were released in the free volume of the column. The eluate is collected and freeze-dried. Get 9 g of the product with a content of nucleotide components 80% abs.v., protein 8% abs.s., lipids 10% abs.v.

The pasty cell mass is washed with water and dried sublimated. Get 58 g of the protein product with a moisture content of 5%, a protein content of 65% abs.s.v., nucleic acids 0.5% abs.s.., spshidov 0.9% abs.s., carbohydrates 30% abs.c.

From the data in the table it can be seen that, when processing fungal biomass under the described conditions, the protein content in the protein product increases by 27% compared with the initial biomass (with

38 to 65%), whereas by a known method this indicator increases by only 18%, the biological value increases by 10% (from 55 to 65%), while by a known method this indicator decreases by

20%. The content of lipids and nucleic acids does not exceed 1% and meets the requirements of the PI AMS SSSR and FAO (WHO).

Microbial fat contains 75% lipids (compared with 35% in the bottom residue obtained by a known method) and is a product suitable for technical use without further purification.

The preparation of nucleotide components contains 80% of nucleotides (compared with 31% in an aqueous extract,

obtained by a known method) and is an intermediate product suitable for use in the chemical industry.

Example 5. In the first stage of alcohol extraction to 100 g of spray-dried biomass of bacteria Acetobacter sp. strain BWA-914 with wet; nost. 5%, protein content 70% abs. s.v ,, nucleic acids 13% abs.c., lipids 9% abs.c., carbohydrates 4% abs.s.., 371 g of ethanol and water are added. The alcohol content in the liquid phase of the system is 75%, the solid phase: liquid phase ratio is 1; 5. The resulting suspension is kept at 30 ° C for 50 minutes with continuous stirring and the cellular product is separated from the alcoholic extract by separation. Get 257 g of pasta with a content of 35% abs.s.v.

In the second stage of alcohol extraction, 733 g of ethanol is added to the paste. The alcohol content in the liquid phase of the system is about 92% of the topic; the solid phase: liquid phase ratio is 1:10. The resulting suspension is maintained at 30 ° C for 50 minutes with continuous stirring and the cellular product is separated from the alcoholic extract by separation.

 The alcoholic extracts obtained in the first and second stages are combined and freed from the solvent by vacuum evaporation. The solvent is reused. After vacuum evaporation, 10 tons of bottom residue are obtained, representing microbial fat with a lipid content of 75%, protein 16%, nucleic acids 4%,

The residue of the extracted cell masses was dried from the solvent by vacuum, and 84 g of defatted product were obtained.

The fat-free product is mixed with 420 ml of an aqueous solution of sulfuric acid with a concentration of 0.1 g-mol / l. The system has a pH of 1.2. Further processing of the suspension is carried out in two series-connected tubular heat exchangers. In the first, the system is heated before and held for 3 minutes, in the second - cooled to 25-30 C. After cooling, the suspension is neutralized with 10% aqueous potassium hydroxide solution pH 4.0 and the solid and the extract separated by centrifugation.

The acidic extract is dried sublimationally and 15 g of the product with the content of nucleic acids of 75% abs.s.v., protein 19% abs.s.., lipid 3% abs.v.

The pasty cell mass of the prod is dried with water and dried sublimated. Get 71 g of the protein product with a moisture content of 5%, a protein content of 89% abs.s., nucleic acid content of 0.4% abs.in., lipids 0.4% abs abs, carbohydrate 4% abs .with.

In the data in the table, it can be seen that when processing bacterial biomass under the described conditions, the protein content in the protein product increases by 19% (from 70 to 89%) compared to the initial biomass, then by a known method this indicator is 18% older, biological value increases by 15% (from 75 to 90%), while by a known method this indicator decreases by 20%. The content of lipids and nucleic acids does not exceed 1% and satisfies the requirements of the PI AMN SSSR and FAO (WHO).

Microbial fat contains 75% lipids (compared with 35% in vat

g

0 5 o

Q

° 5

five

five

the residue obtained by a known method) and represents a product suitable for technical use without further purification,. The preparation of nucleotide components contains 75% of the nucleotides (compared to 31% in the aqueous extract obtained by a known method) and is an intermediate product suitable for use in the chemical industry.

Example 6. The table shows the treatment conditions for spray-dried biomass of Acetobacter sp. strain BWA-914,

The alcohol extraction is carried out with isopropanol and lipid components are isolated only from the alcoholic extract obtained in the second stage of alcoholic extraction.

Extraction of nucleic acids is carried out with an aqueous solution of hydrochloric acid and neutralized with calla hydroxide. Acid extract is spray dried. The cell mass is washed with water and dried, the sublimation

From the data in the table it can be seen that in the first and second stages of alcohol extraction, the concentration of alcohol is lower than what is required by the proposed method. This leads to the fact that the protein product contains an excess of (5.8%) the amount of lipids, and its biological value is not yBejm-.

In addition, the distillation residue after distillation of the solvent contains 32% protein and 40% lipids, which cannot be allocated as a preparation of lipid components.

Thus, the purpose of the invention is not achieved under these conditions.

Example 7. The table shows the treatment conditions for spray-dried biomass of the yeast Candida maltosa strain BWA-777,

The alcohol extraction is carried out in one stage for 120 minutes at 40 ° C with methanol, and the lipids are removed from the extract.

Extraction of nucleic acids is carried out with an aqueous solution of phosphoric acid at an overpressure (1 bar). The acid extract without neutralization is ultrafiltered in Example 2, the rethettate is washed with water and dried sublimation.

Cell masses are washed with water and freeze-dried.

From the data in the table, it is clear that alcohol extraction is carried out in one stage I with an alcohol concentration greater than that required by the invention, and the acid extract is separated without neutralization. This leads to the fact that the protein B content of the protein product is only: 63% and does not reach the value (65%) required by the norms of the AMS USSR and FAO (WHO). The biological value of the protein product is reduced by 5% compared with the original biomass (from 70%: to 65%).

; In addition, the acidic extract pos: le ultrafiltration contains 45% of protein and only 38% of nucleotides that cannot be isolated as a prepara- tory of nucleotide components. Thus, in these conditions

From the data in the table it can be seen that the acid extraction is carried out at a pH higher than that required by the present invention. This leads to the fact that the protein product contains an excessive amount of the nucleus of the invention is not achieved.

Example 8. The table shows 25 leic acids (2.6%) and lipids treated by spraying (1, -3%), the biological value of the protein product does not increase.

Thus, in these conditions, the purpose of the invention is not achieved. 30 EXAMPLE The table shows the biomass processing conditions for the yeast Candida maltosa strain BWA-777.

The alcohol treatment is carried out with methanol and the lipid components are isolated from the combined alcoholic biomass of the Candida maltosa yeast strain BWA-777.

The alcohol treatment is carried out with ethanol. Lipids are isolated from the combined alcoholic extract.

Extraction of nucleic acids is carried out with an aqueous solution of phosphoric acid and neutralized with potassium hydroxide. The acidic extract is ultrafiltered according to example 2. The retestat is washed with water and dried by sublimation.

Cell masses are washed with water and dried.

From the data in the table it can be seen that the acid extraction is carried out at a pH lower than that required by the present invention. This leads to the fact that the protein content in the protein product is only 62% and does not reach the value (65%) required by the standards of the AMS USSR and FAO (WHO). The biological value of the protein product is reduced by 2% (from 70 to 68%).

In addition, the acid extract after ultrafiltration contains 30% protein and 43% nucleotides, which cannot be isolated as a preparation of nucleotide components.

Thus, in these conditions. The purpose of the invention is not achieved.

Example 9. The table shows spray processing conditions.

35

40

rakta

The extraction of nucleic acids is carried out with an aqueous solution of hydrochloric acid, neutralized with sodium hydroxide and spray dried.

Cell masses are washed with water and freeze-dried.

50

From the data in the table it can be seen that the acid extraction is carried out for a shorter period of time than is required by the present invention. This leads to the fact that the protein content in the protein product is only 59%, which is significantly lower than the norm required by the AMS USSR and FAO (WHO), and the content of nucleic acids (9%) and lipids (5%) significantly exceeds these norms. . The biological value of the protein product does not increase.

In addition, the acidic extract contains 30% protein and 40% nucleotides, which cannot be isolated in viruses.

dried biomass of bacteria Acetobacter sp. strain BWA-914.

The alcohol treatment is carried out with isopropanol and the lipid components are extracted only from the alcohol extract. that obtained in the second stage of alcohol extraction.

The extraction of nucleic acids is carried out with an aqueous solution of phosphoric acid and neutralized with tri-sodium phosphate.

The acidic extract is subjected to ultrafiltration according to Example 2. The rethettate is washed with water and freeze-dried.

I "

Cell masses are washed with water and

dried by sublimation.

From the data in the table it can be seen that the acid extraction is carried out at a pH higher than that required by the present invention. This leads to the fact that the protein product contains an excessive amount of nucleic acids (2.6%) and lipids (1, -3%), the biological value of the protein product does not increase.

The alcohol treatment is carried out with methanol and the lipid components are separated from the combined alcohol extract

rakta

The extraction of nucleic acids is carried out with an aqueous solution of hydrochloric acid, neutralized with sodium hydroxide and spray dried.

Cell masses are washed with water and freeze-dried.

From the data in the table it can be seen that the acid extraction is carried out for a shorter period of time than is required by the present invention. This leads to the fact that the protein content in the protein product is only 59%, which is significantly lower than the norm required by the AMS USSR and FAO (WHO), and the content of nucleic acids (9%) and lipids (5%) significantly exceeds these norms. . The biological value of the protein product does not increase.

In addition, the acidic extract contains 30% protein and 40% nucleotides, which cannot be isolated in vitro.

de drug nucleotide components.

Thus, in these conditions, the purpose of the invention is not achieved.

Example 11. The conditions for the treatment of the biomass of the bacteria Acetobacter sp. Are shown in the table. strain BWA-914,

The alcoholic treatment is carried out with isopropanol and lipid components are isolated only from the alcoholic extract obtained in the second stage of alcoholic extraction.

The extraction of nucleic acids is carried out with an aqueous solution of sulfuric acid and neutralized with an aqueous solution of potassium hydroxide. Acid extract is freeze-dried. Cell masses are washed with water and freeze-dried.

From the table, it is apparent that acid extraction is carried out for a longer period of time than is required by the present invention. This leads to the fact that the biological value of the protein pro. The duct does not increase.

In addition, the acid extract contains A0% protein and 40% nucleotides, which cannot be isolated as a preparation of nucleotide components.

. Thus, in these conditions, the purpose of the invention is not achieved.

Example 12. The table shows the processing conditions of the yeast Candida maltosa strain BWA-777.

The alcohol treatment is carried out with ethanol and the lipid is isolated. components I8 combined alcoholic extract.

The extraction of nucleic acids is carried out with an aqueous solution of phosphoric acid under overpressure (1 bar) and neutralized with an aqueous solution of sodium hydroxide. The acidic extract is subjected to ultrafiltration according to example 2. The retettat is washed with water and dried by sublimation. Cell masses are washed with vrdu and spray dried.

From the data in the table it can be seen that the acid-free extraction is carried out at a temperature higher than that required by the invention. This leads to the fact that the protein content in the protein product is only 59%, which is significantly below the norm required by AN AMS USSR and FAO.

Yu

is 20

25

"

five

0

five

0

(WHO). The biological value of the protein product is reduced by 5% (from 70 to 65%).

In addition, the acid extract after ultrafiltration contains 35% protein and 44% nucleotides, which cannot be isolated as a preparation of nucleotide components.

Thus, in these conditions, the purpose of the invention is not achieved.

Example 13. The table shows the processing conditions of the bacteria Acetobacter sp. strain BWA-914.

The alcohol treatment is carried out with isopropanol and the lipid components are isolated only from the alcohol extract obtained in the second stage of the alcohol extract.

The extraction of nucleic acids is carried out with an aqueous solution of hydrochloric acid and neutralized with an aqueous solution of sodium hydroxide. Acid extract is spray dried. Cell masses are washed with water and spray dried.

From the data in the table, it can be seen that the acid extraction is carried out at a lower temperature than that required by the invention. This leads to the fact that the protein product contains an excessive amount of nucleic acids (4.1%) and lipids (1.4%). The biological value of the protein product does not increase.

Example 14. The table shows the processing conditions of the yeast Candida maltosa strain BWA-777.

The alcohol treatment is carried out with ethanol and the lipid components are isolated from the combined alcohol extract. The extraction of nucleic acids is carried out with an aqueous solution of phosphoric acid and neutralized with an aqueous solution of potassium hydroxide. Acid extract is subjected to ultrafiltration according to example 2. Retettat is washed with water and dried by sublimation. Cell masses are washed with water and dried by sublimation.

From the table it is seen that the acid extract is neutralized to a higher value than is required by the present invention. This leads to the fact that the protein product contains only 62% of protein, which is significantly below the norm required by the USSR Academy of Medical Sciences and FAO (WHO). The biological value of the protein product does not increase.

In addition, the acid extract after ultrafiltration contains 40% protein and 35% nucleotides that are not. can be isolated as a preparation of nucleotide components.

Summing up the data in the table, we can conclude that only the processing of yeast, bacterial and fungal biomass with alcohols in two stages with the concentration of alcohol in the liquid phase of the system in the first stage is 50-75% and

In addition, there is no complex processing, i.e. it is not possible to obtain a preparation of lipid components.

When carrying out the acid extraction process under milder conditions, for example, at higher pH or lower temperatures (50 ° C), a protein product is obtained with an excess of lipids and nucleic acids and low biological value.

Carrying out the acid process

in the second stage, 80–96%, followed by extraction for a longer period — treatment of the defatted product in water time (5 min) leads to acid solutions at a pH of 0.7–1.5 for 1–5 min at 50 -100 ° C and neutralized to pH 2.520

4,5 allows you to complexly obtain a protein product with a protein content of more than 65% and a content of nucleic acids and lipids not exceeding 1%, and higher biological value, as well as a preparation of lipid and 25 nucleotide components with a basic substance content of at least 70%. process in more severe conditions, such as alcohol extraction of protein product with low biological value, in addition, there is no complexity of processing, namely, it is impossible to isolate the preparation of nucleotide components.

Claims (1)

Invention Formula The method of purification of biomass of microorganisms, including the processing of its aqueous solutions of lower alcohols with the conduct of alcohol extraction. 35 separation in one stage or higher, the separation of the alcoholic extract and the alcohol content (75%), acidic extraction at a lower pH (0.7) / or higher temperature (100 ° C) results in a protein product low protein and low biological value. In addition, under these conditions it is impossible to carry out the process in a complex, namely, to isolate the preparation of nucleotide components, Similar results are obtained by carrying out acid treatment for a shorter time (i = 1 min) or neutralization to a higher pH (1.5), When carrying out alcohol extraction at a lower concentration of alcohol (50%) in the liquid phase of the system, a protein product with an excess of lipids, low, g, is obtained. 40 45 removal of alcohol residues from it, followed by water extraction and separation of the aqueous extract, characterized in that, in order to improve the quality of the bak product while obtaining lipid and nucleotide components, alcohol extraction is carried out in two stages with alcohol in liquid the phase of the system in the fifth stage is 50-75% and in the second 80a the resulting alcohol extrusions are then combined and the liquid components are made of them, then the degreased material is treated with aqueous solutions of acids at pH 7.0-1.5 for 1- 5 min with that erature 5100 ° C, neutralized to pH 2.5-4.5 and from the extract obtained vschel nucleotide components. extraction over a longer period (5 min) leads to obtaining a protein product with a low biological value, in addition, there is no complexity of processing, namely it is impossible to isolate the drug nucleotide components. Invention Formula The method of purification of biomass of microorganisms, including the processing of its aqueous solutions of lower alcohols with the conduct of alcohol extraction. separation of alcoholic extract and removal of alcohol residues from it, followed by aqueous extraction and separation of the aqueous extract, characterized in that, in order to improve the quality of the protein product while obtaining lipid and nucleotide components, the alcohol extraction is carried out in two stages with an alcohol content in the liquid phase of the system the first stage is 50-75% and the second is 8096%, and then the resulting alcoholic extracts are combined and the lipid components are made of them, then the fat-free material is treated with aqueous solutions of acids at pH 7.0-1.5 volts for 1-5 minutes at a temperature of 50-100 ° C, neutralized to a pH of 2.5-4.5, and nucleotide components are extracted from the obtained extract. OM g about o 00 "3 oo t OfOO OOOtO h- - h C -f «-G-h-in -Sh- - to az e; saasiuazs; Msec; sa s; shave; shavs; a tn 9 r I About About HF 1G | “P so O -. ShSO1L ol1 "- MO1L  "- fc  Osmo evO "ri 0" 1L emolo tnmoo -c4OvO Oin - sog. "-) - with  iftine .г.о 1О c4ir) en oo oo oo CO m SO - -.00 r-J § S about ate 1L g 1L C1 so1L "Sg sh g you sh 1L one about djlo OO §3 PM  g s sh W II 31 o s to f W o L w o (W to I o "9 o I.S Shh & e "1" ia iiua And h he 71 & . CU. Co and m p with; and s h "  ".§ S 114 yu aia gg oioi about ., about 4J H.U o e g i §five yu a 1G1 - chl Jk (h sh “R | about m oh oh i about sh oh oh S Oh oh about Yu about BUT P about in about m sh "L about " about about k § lo o cm Oh ev g "m ON about 00