SU908085A1 - Method for preparing biomass - Google Patents

Abstract

METHOD OF OBTAINING BIOMASS, involving the cultivation of methacid-producing bacteria, including Methylococcus capsulatus, under non-sterile conditions on a nutrient medium containing sources of nitrogen, phosphorus, mineral salts and methane as a carbon source, while maintaining a constant concentration of one of the elements of the nutrient medium during cultivation characterized in that, in order to simplify the process and increase productivity, the concentration of copper ions in the cultivation process is from 0.05 to 2 mg / l. (L with

Description

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The invention relates to the microbiological industry, and HNieHHO to a method for producing biomass of methacidic bacteria.

It is known THAT: certain concentrations of salts in the nutrient medium when growing certain types of methane-acidifying bacteria have an inhibitory effect on the growth of these microorganisms fj.

. It is also known that pure cultures of Methylococcus capsulatus grow well in the presence of copper salts in the nutrient medium {e.

A biomass production method is known, involving the cultivation of methacid-forming bacteria, including Methylococcus capsulatus, in non-sterile conditions on a nutrient medium containing sources of nitrogen, phosphorus, mineral salts and methane as a carbon source,. while maintaining a constant concentration. one of the elements of the nutrient medium in the process of cultivation h.

According to this method, the concentration of ammonium ions is kept constant. In this process, carried out in several stages, and the maximum productivity of the process, achieved with an increase in pressure in the fermenter from 0.35 MPa, is 5 g / l / h

The purpose of the invention is to streamline the process and increase productivity.

The goal is achieved by the fact that, when implementing the method of producing biomass, it is necessary to cultivate methane-acidifying bacteria, including Methylococcus capsulatus, under unsterile conditions on a nutrient medium containing sources of nitrogen, phosphorus, mineral salts and methane as a carbon source, while maintaining a constant concentration of nutrient media during cultivation maintain the concentration of copper ions from 0.05 to 2 mg / l.

The method is carried out as follows.

The cultivation of methane-oxidizing cultures of microorganisms, including Methylococcus capsulatus, is carried out in an aqueous mineral saline nutrient medium under conditions of aeration and re-measurement with a continuous supply of gases containing methane and oxygen.

In this case, various mixed methane-oxidizing cultures can be used, including the bacterium G Methylococcus capsulatus, for example, such as the strain 1-70 containing the bacteria Methyloceus capsulatus, Methylosinus sporium Methylosinus trichosposium, Flavobacterium gasptypicum.

The SOURCE of nitrogen can serve as ammonium or other nitrogen salt, TaKjj ammonia water, which can be.

use both as a source of nitrogen and as a titrating agent to stabilize the pH in the process. Phosphoric acid or phosphoric acid salts are used as a source of phosphorus, and potassium, magnesium sources, and trace elements such as iron, zinc copper, m-arganese, and others are served as salts in the form of salts as mineral sources of microorganisms. maintain the concentration of copper ions in the culture fluid in the range of 0.05-2, preferably 0.1-1.0 mg / l, creating conditions for the growth of the most active producer of Methanococcus capsulatus, the methane-oxidizing bacteria and the oppression of the development of bacteria Utniks. Growing methanogenic mixed cultures is carried out at a pH of 5.06, 5, temperature ZO-BO C and flow rates of 0.25-0.33 hours. The final culture is dominated by bacteria Methylococcus capsulatus whose content is 85-95% .

Cultivating is preferably carried out under increased pressure.

Example. Mixed cultures of methane-acidifying microorganisms, sotots of the bacteria Methylococcus capsulatus and Methvlosinus sporium were grown on nutrient medium of the following composition, g / l:

HjPO (80%)

17.2 4 5

MgSO.-THnO KCl

Oh, Ob

ZnSO -lEqO MpZO. And HNO 0.38 0.25,: 0.21

FeS047H20 0., 009 IalMoO —H o

0,0095

Ammonia water was supplied as a nitrogen source and titration agent. In this case, a concentrated solution of CuS045H2O was fed separately to maintain the concentration of ions in the culture media to be 0.05 mg / l. The process was carried out at a temperature of 33-40 C, pH 5.4-5.5 and continuous supply of the gas mixture, with natural gas in the amount of 10 l / l of medium / h and air in the amount of 30 l / l of medium / h.

Upon reaching a biomass concentration of 20 g / lasv, a continuous cultivation process was started with a dilution factor of d 0.25 h. The concentration of copper in the culture medium was regularly checked and corrected.

Bacteria Methyloc occu. capsulatus in mixed culture was 85% at a constant biomass concentration of 20 g / l DIA. The resulting biomass was separated by separation from the culture fluid and dried. EXAMPLE 2 A mixed culture of methanol acid to bacteria dichloride, consisting of bacteria Methylococcus sarsulatus, Methy3.os inus .sporium, Methylosinus trichosporium, was grown in non-sterile conditions at pH 5.8-6.0 and temperature 43- 45 ° C, the number and composition of the supplied gas mixture and nutrient medium are the same as in example 1. The concentration of copper ions in the culture medium was maintained at 2 mg / l. Upon reaching a biomass concentration of 10 g DIA / L, the pressure in the fermenter was gradually increased to 0.7 MPa. With increasing pressure, the supply of gaseous and liquid sources of microorganisms was increased. The residual concentration of honey in the culture fluid was regularly checked. When the concentration of copper decreased below 2.0 mg / l, a solution of copper salts was added. With “an increase in the copper concentration above 2 mg / l, the supply of the copper salt solution was reduced or stopped for a while. The concentration of biomass at a specified flow rate of 0.27 was equal to 37 g / l DIA, and the degree of dominance of Methylococcu capsulatus in the mixed culture was 85%. The resulting biomass was separated by separating from the culture fluid and dried. PRI me R 3. A mixed methane-culture culture was grown, including the following bacteria: Methylococcus capsulatus, Methylosinus sporium, Methylosinus trichosporium, Flavobacterium “asotypicum. The process was carried out at a pH of 5.3-5.4 and a temperature of 40-42 0. Copper concentration of 0.1 mg / l was maintained in the culture liquid. The composition of the supplied liquid nutrient medium and gae mixture is the same as in Example 1. Fermentation was carried out under a pressure of 0.9 MPa. The process indicators were as follows: 3–3 a biomass concentration of 50 g / l DIA, the degree of dominance of Methylococcus capsulatus bacteria in a mixed culture was 95%. The obtained biomass was separated by separation from the culture liquid and dried. PRI me R 4. The mixed methane-acid culture was grown by the analogous culture in example 3. The process was carried out at pH 5.5-5.6 and temperature 42-43 ° C. The concentration of copper in the culture liquid was 1 m. The process was conducted under a pressure of 0.9 MPa. The composition of the liquid nutrient medium, the gas mixture supplied to the apparatus as in the previous examples. Process indicators were as follows: d .Of33 h, biomass concentration 45 g / l DIA, the degree of dominance of Methylococcus capij-alatus bacteria in a mixed culture was 95%. The resulting mass was dehydrated and dried. Example5. A mixed methanoacid was grown for a whole culture similar to that in Example 3. The process was carried out at a pH of 5.6-5.7 and a temperature of 43-44c. Copper concentration in the culture liquid was maintained at about 5 mg / l. The process was conducted under pressure of 0.9 MPa. The composition of the liquid nutrient medium and the gas mixture supplied to the apparatus, as in the previous examples. The process indicators were as follows: d 0.3 The concentration of biomass was 50 g / l DIA, the degree of predominance of Methylococcus caosulatus bacteria in the mixed culture was 95%. The ddJ biomass was separated by separa-j from the culture fluid and dried ... EXAMPLE 6 (comparison with prototype). A mixed methacid culture consisting of bacteria jMethylococcus capsulatus, Methylosinus sporium, Methylosinus trichosporium. Flavobacteriura gasotyp / cum, was grown on a nutrient medium similar to the medium in Example 1. Ammonia water was supplied as a source of nitrogen and a titrating agent. The concentrated solution of 5H 2 O was fed separately in the amounts necessary to maintain the concentration of copper ions in the culture medium equal to 1.5 mg / l. The process was carried out at a temperature of 42-45 ° C, pH 5.6-57 and continuous supply of a gas mixture consisting of methane natural gas in an amount of 10 l / l of medium / h and air in an amount of 30 l / l of medium / h. When the biomass concentration reached 10–15 g / l DIA in a continuous process with a dilution factor d 0, the pressure in the fermenter was gradually increased to 0.35 MPa. With increasing pressure, the supply of gaseous and liquid microbial power sources increased proportionally. The residual concentration of copper in the culture medium was regularly monitored. When the pressure in the fermenter was 3.5 kg / cm, the process parameters were as follows: d 0, 20 h, the biomass concentration was 30 g / l DIA, the productivity of the process was 6 g / l / h. The content of bacteria Methylococcus capsulatus In a mixed culture; LO was stable and was 90%. / The proposed method allows to simplify the process of obtaining biomass.

908085

methanol-oxidizing bacteria in non-sterile, high protein content, good conditions due to the maintenance and productivity in the process of cultivation of a constant process due to the possible concentration of copper ions, to obtain the cultivation of the biomass, which is dominated; with a high rate of Rosbacterin Methylococcus capsulatus.

Claims (1)

METHOD FOR PRODUCING BIOMASS, involving the cultivation of methane-oxidizing bacteria, including Methylococcus capsulatus, in non-sterile conditions on a nutrient medium containing sources of nitrogen, phosphorus, mineral salts and methane as a carbon source, while maintaining a constant concentration of one of the elements of the nutrient medium during cultivation, characterized in that in order to simplify the process and increase productivity, the concentration of copper ions from 0.05 to 2 mg / L is maintained during the cultivation process.