SU686459A1 - Method of producing biomass of microorganisms - Google Patents

Abstract

THE METHOD OF OBTAINING BIOMASSYMICROORGANISMS by growing the bacteria of the genus Methylomonas producing it on a nutrient medium containing methane as a carbon source, the source: nitrogen, necessary mineral salts in the presence of trace elements in the form of iron and copper ions during aeration of the environment, characterized in that, in order to increase biomass yield, from bacteria of the genus Methylomonas, Methylomonas methanica strain 12 is used, and cultivation is carried out on a nutrient medium containing calcium, zinc, manganese, molybdenum and icobalt ions as additional and source of micronutrients.

Description

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about:

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The invention relates to the microbiological industry, namely to a method for producing biomass of microorganisms using methane as a source of carbon and energy.,.

Methane is one of the cheapest and most affordable carbon sources for the synthesis of microbial biomass which can be used for feeding purposes, in various areas of industry, for reducing the methane content of coal seams, etc. The essential advantage of the process of obtaining biomass using methane-acidifying microorganisms is the absence of a stage of purification of the final product from the unutilized carbon substrate.

A known method for producing biomass of microorganisms by growing bacteria of the genus Methylomonas producing them on a nutrient medium containing methane as a carbon source, nitrogen source, essential mineral salts in the presence of trace elements in the form of iron and copper ions during aeration of the medium.

The known method provides the maximum productivity of the process of continuous cultivation of bacteria of 0.645 g / l — with a biomass concentration of 4.3 g / l and a dilution rate of 0.15 h.

The disadvantage of this method is the low biomass yield.

The purpose of the method is to increase the biomass yield of bacteria of the genus Methylomonas

The goal is achieved by the use of Methylomonas methanica strain 12 from bacteria of the genus Methylomonas, and cultivation is carried out on a nutrient medium containing calcium, zinc, manganese, molybdenum and cobalt ions as additional sources of microelements.

The strain Methylomonas methanica 12, isolated from a sample of lake sludge from the vicinity of the town of Pushchino, is stored in the crop collection of the laboratory of biogeochemistry of the Institute of Biochemistry and Physiology, Microorganism of the Academy of Sciences of the USSR.

Characterization of Methylomonas methanica strain 12

1. Morphological and cultural properties:

86459

-palocarpous bacteria, size 0.5-0.7 1.0-2.5 microns, motile, due to the presence of flagella;

-Game-negative;

J - possesses: type 1 intracytoplasmic membranes of the structure in the form of stacks of closed vesicles;

- the river socket does not form;

- sometimes form immature cysts to type Azotobacter;

- do not possess a capsule;

2. Physical and biochemical properties:

-strict aerobes;

- growing temperature range 2015

- pH range 5.5-9.0;

- use methane or methanol as a source of carbon and energy;

20 - use carbon dioxide as a carbon source; :

- do not grow on environments with formaldehyde, formate, carbon monoxide, ethane, propane, butane,

25 alcohols (normal and isomers), methylated amines, methyl urea, malate, acetate, succinate, glucose, on MPA, MPB, wort, glucose-potato agar;

30 - use nitrate or ammonium salts as a source of nitrogen;

- do not use nitrites, methylated amines and methyl urea in

ee as a nitrogen source;

-assimilate with carbon of methane or methanol at the level of formaldehyde in the ribulose-phosphate cycle;

- have the activity of rat dehydrogenase (NAD and NADP-specific), glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase;

- do not possess the activity of oxy45 pyruvate reductase, serine glyoxylate transaminase and d-ketoglutarate dehydrogenase.

Example 1. The cultivation of Methylomonas methanica strain 12 50 is carried out on an aqueous nutrient medium of the following composition, g / l:

0.4

2H20

0.4

0.8

-3

80-10

40-107НГО

4.0-10-3 5Nur

2.0-10-3

7HjO

SnSO /

10-3 5HjO

Mp1g

1.2 10-5

NaiMoO ,, 2НгО 1,2

0.4-10

0.2 ML / L

HiSQj, END.

pH of the medium 2.6

The medium is sterilized by autoclaving at a temperature of 1 hour.

A 200 ml suspension of Methylomonas methanica strain 12, grown in flasks on a shaker at 30 ° C, was added to the fermenter Biotec FL103 manufactured by LKB, Sweden, and 400 MP of the medium and 400 ml of distilled water were added. The concentration of cells in the menter is 0.1 g / l. The temperature in the fermenter is maintained; the rotational speed of the stirrer is 1500 rpm, pH 6.2. The supply of nitrogen and the stabilization of the pH are ensured by the automatic titration of the medium with a 5% solution of Naj, OH.

The culture fluid is sparged with a gas mixture consisting of: natural gas and air at a ratio of 1: 4 at a rate of 1 l / l-min.

For 24 hours, the cultivation is carried out in a batch mode, then the process of continuous cultivation begins.

The concentration of biomass under established conditions, corresponding to different dilution rates, is shown in the table.

With continuous cultivation of Methylomonas methanica strain 12, the maximum productivity of the process is reached, equal to 1.26 g / l "h" with a biomass concentration of 9.0 g / l and a dilution rate of 0.14 hH

Thus, the biomass yield is twice the yield obtained using the method described in the prototype of the proposed invention.

Biomass concentration

g / l 11.4 10.2 9.0

6.5

7,8

Claims (1)

METHOD FOR PRODUCING MICROORGANISM BIOMASS by growing bacteria of the genus Methylomonas producing it on a nutrient medium containing methane as a carbon source, source: nitrogen, necessary mineral compounds in the form of trace elements in the form of iron and copper ions during aeration of the medium, characterized in that, in order to increase yield of biomass from bacteria of the genus Methylomonas use Methylomonas methanica strain 12, and the cultivation is carried out on a nutrient medium containing calcium, zinc, manganese, molybdenum and cobalt ions as additional micronutrient Source. S $ ω s: Q0 Σ © <686459 2