SU1076446A1 - Strain pseudomonas putida 36 for use in purification of water and soil from petroleum and petroleum products - Google Patents

Abstract

The strain Pseudomonas putida 36. (deposited in the Central Museum of Industrial Microorganisms of the Institute of Scientific Research Institute of Genetics, collection number CMPM B-2443), used to purify water and soil from oil and oil products. (L / S1 Od 4 4 yes f 8 Time, ffH

Description

This invention relates to the microbiological industry and relates to a dance used in the biological treatment of water from oil products and soil from residual soil.

A known strain of pseudomonas aeruginosa RAO, containing plasmidases CAM / OCT, NAH, SAL, controlling the utilization of hydrocarbons, used, l microbiological degradation of petroleum and petroleum products l;

The disadvantage of the multi-plasmid tamm Pseudomonas aeruginosa RAW is the low rate of recovery of crude oil.

Also known strain Pseudomonas aeruginosa 121, capable of utilizing octane 2.

The disadvantages of this strain are the low utilization rate and the narrow spectrum of selective action.

The closest in terms of the achieved result to the proposed is Tam Pseudomonas aeruginosa 123, carrying the EAST, CAM NAH plasmid, used in the biological purification of water from oil and oil products

The residue of a known strain is a narrow spectrum of selective action; it resorbs only the oil film., it is not able to dispose of residual soil contamination. The purpose of the invention is. obtaining a strain capable of carrying out degradation of oil, residual oiling, which can be used in the purification of water and soil from pollution by oil products.

The resulting strain Pseudbmonas putida 36 was isolated from an organic based, spent (oil) drilling mud stored in a sludge-pit open pit bore (well 13100, bush 1030, Belozersk oil field of the Tyumen region, October 13, 1980) and deposited in VNIIgenetics , collection number CMPM B-2443.

The obtained strain of Pseudomonas putida 36 (collection of the museum of microbiology at the laboratory of the Ministry of Medicine and Social Sciences of Russia) has the following characteristic.

Morphological and cultural characteristics. The size of the cells of a one-day culture of 1.7-2.4x0.3-0.5 microns. The predominant form of cells is straight, rod-shaped. Gram-negative. . On m of septon agar, colonies are convex, round, colorless. Ryzer 3-7 mm.

M no-septone broth - a turbidity is formed with gas evolution. Potatoes plentiful growth, a stroke convex, continuous, with formation of slime. Gelatin does not dilute. Milk coagulates, acid.

Physiological signs. Azrob. 5 Grows at 28-42 ° С, does not grow at 4 ° С.

The ratio is to carbohydrates. Uses lactose, glucose and maltose gas. Does not absorb arabinose. 0 Relation to hydrocarbons. Uses naphthalene, camphor, octane, hecaiH, benzene, toluene, xylene, paraffin, asphaltenes. Paraffinic oils assimilate more actively than aromatic ones.

Attitude to food sources. Assimilates nitrate nitrogen.

The drawing shows a graph showing the biomass growth curves of the Q strains of P. putida 36 and P. aeruginosa 123 on crude oil.

Example. The strain Pseudomonas putida 36 is grown on the mineral medium of the following composition, g: 5 KjHPO ,, 10.0

KN ROts1,0

2.0

NH xNO, "2

1000.0

 A ml flask is added with 50 ml of mineral medium, 2.5 ml of a hydrocarbon feed source (crude oil) and 1.5-10 cells. The cultivation is performed on a rocking chair at. Samples are taken daily, the number of viable cells 5 is determined by seeding the dilutions on MI1A medium. The counting of the colonies is carried out after 24 hours. The oxidative activity of the strain is determined by the amount of carbon dioxide emitted. 0 In the graph, the curves of oxidative activity of the strains Pseudomonas putiida 36 and Pseudcanonas aeruginosa 123 on crude oil demonstrate different. Ability of these strains to degrade crude oil.

Curve a shows the change in cell density (the number of bacterial cells in 1 ml of the strain medium) of the strain Pseudomonas aeruginosa 123 with time at an oil concentration of up to 5% in the medium.

Curve 5 shows the biomass growth of the strain P. putida Zb. The maximum density (1.510 cells / ml) strain P, putida 36 reaches 56 days, and the strain P „aeruginosa 123 reaches a maximum density of 1.5 aokl / ml 2 -4 days

The oxidative activity of the proposed and known strains on petroleum is presented in the table.

264.0 240.5 74.5 68.4

145.2 187.0 33.7 37.1

As can be seen from the table, at a practically equal growth rate, the oxidative activity of the strain P.putida 36 on crude oil significantly exceeds the oxidative capacity of the strain P. aeruginosa 123. Thus, the strain P. putida 36 isolates

oxidation of paraffinic and aromatic oils, respectively, 264, and 240.5 COg mg per 100 g of soil, and the strain P. aeruginoJsa 123-145.2 and 187.0 mg COg, respectively.

These tables show that for 30 days the amount of oil utilized by the strain p. putida 36,

. reaches 68.4-74.5% of the original, and P. P. aeruginosa 123 33.737, 1%.

Thus, the advantage of the proposed strain P. putida 36 is the ability to grow even in a depleted medium (water with mineralization not higher than 10% can be used), the ability to utilize paraffin, naphthalene, octane hexane, benzene, toluene, xylene, and: sphaltenes, as well as the ability to degrade oil and oil products at high speed with oil content up to 10% by volume of contaminated

substrate (soil, water), which allows its use in the biological purification of water from oil and soil from residual oil content.

Claims (1)

Thus, the advantage of the proposed strain / R. Ribtba 36 is the ability to grow even in a depleted medium (produced water with a salinity not higher than 10% can be used), the ability to utilize paraffin, naphthalene, octane, hexane, benzene, toluene, xylene, asphaltenes, and also the ability to degrade oil and oil products at high speed with oil content up to 10% of the volume of contaminated substrate (soil, water), which allows its use in the biological purification of water from oil and soil from residual earth formation.