RU2476385C1 - Method of sewage water purification from phenol compounds - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to field of biochemistry. Sewage water with increased content of phenol compounds 300 - 1200 mg/l, content of sulfide-ions to 64.1 mg/l and content of sulfate-ions to 401.4 mg/l and 1843.7 mg/l in case of sparging with air, is passed through column type reactor. Reactor contains adsorbent - petroleum coke, onto which cells of strain of aerobic bacteria Pseudomonas putida 131 All-Russian collection of industrial microorganisms B-10894 are immobilised. Orthophosphoric acid or its salts are preliminarily dissolved in sewage water as biogenic additive in order to obtain pH 7-8, optimal for phenol destruction by bacteria strain.

EFFECT: invention ensures increased quality of purification of sewage water, which contain sulfide-ions and sulfate-ions, from phenol compounds to 0-50 mg/l.

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Description

The invention relates to biotechnology, namely to biochemical wastewater treatment from phenolic compounds.

A known method of purification of wastewater from phenols by extraction in the presence of hydrocarbons [A.S. USSR 806616, application 2357297 from 05.10.76], however, this method leads to secondary pollution of wastewater and has high rates of residual phenol.

A known method of biological treatment of wastewater from phenolic compounds using enzyme preparations isolated from homogenates of plant tissues [A.S. USSR 939407, application 2966341 from 09.06.80]. The disadvantage of this method is the complexity of the preparation of enzyme preparations by washing the plant tissue homogenate with acetone or rubbing in phosphate buffer.

A known biological method of wastewater treatment from phenolic compounds using the enzyme preparation o-diphenyl oxidase immobilized on soot [A.S. USSR 914507, application 2966313 dated 06/11/80], but it only affects the stage of biooxidation of pyrocatechol, which is a metabolite in the intermediate oxidation of phenol by bacterial strains.

A known method of biochemical wastewater treatment from phenols by Botrytis cinerea and Phytoptora infestans fungi [A.S. USSR 912684, application 2856399 from 12.20.79].

A known method of biochemical wastewater treatment from phenolic compounds at pH 6-8.5 for 1-3 days using a wood-destroying fungus - white rot Polyporus versicolor [A.S. USSR 969684, application 3277087 from 04.20.81].

A known method of biochemical purification of pre-diluted wastewater from organic compounds by cultivating Candida yeast microorganisms under aeration conditions followed by separation of the microorganisms [A.S. USSR 975588, application 3006003 from 01.08.80].

It is known the use of strain Pseudomonas aeruginosa No. 228, decomposing phenol [A.S. USSR No. 499227, application No. 203990 from 01.07.74].

A known method of biochemical wastewater treatment of phenols with a 3-4-day inoculum of the lignilithic fungus Panus (Lentinus) tigrinus VKM F-3613D [RF Patent 2345957, application 2007123842/13 of 06.25.2007].

A known method of wastewater treatment from phenolic compounds using a bacterial strain Mycobacterium stegmalis VKM B-1205 [A.S. USSR 1597384, application 4368424 from 10.23.87].

A known method of biochemical wastewater treatment from phenol-containing compounds using a strain of microorganisms Pseudomonas aeruginosa XP-25 and in the presence of spatially-hindered alkyl phenols deposited under the number VKPM B-8613 [RF patent No. 2270807, application No. 2004110627 from 04/07/2004].

The above methods of purification are time-consuming, time-consuming and provide a low degree of wastewater treatment from phenolic compounds, in particular from phenolic compounds contained in sulfide-alkaline effluents of oil refineries and petrochemical plants having a high content of sulfide ions. In addition, individual isolated strains are opportunistic and toxic.

The closest in technical essence and the achieved technical result is a method of biochemical wastewater treatment from phenol-containing compounds using a microorganism strain Pseudomonas aeruginosa XP-25, deposited under the number VKPM B-8613 [RF patent No. 2270805, application No. 2004110625 of 04/07/2004]. However, the known strain decomposes phenol in wastewater from the production of phenolic antioxidants or similar in composition, which differs sharply from the composition of sulfide-alkaline effluents of the petrochemical and oil refining industries.

The objective of the invention is to improve the quality of wastewater treatment, in particular sulphide-alkaline effluents of petrochemical and oil refineries, with a high content of sulphide ions, from phenolic compounds, to increase the safety during strain operation, to expand the arsenal of microorganism strains used for wastewater treatment.

The problem is solved by biochemical wastewater treatment using a strain of Pseudomonas putida 131, previously grown on phenol and sorbed on calcined petroleum coke. The strain Pseudomonas putida 131 was isolated from phenol-contaminated soils of the Gazprom Neftekhim Salavat Oil Refinery by cultivating the soil sample in a mineralized medium and then selecting the most active forms and deposited in the All-Russian collection of industrial microorganisms under the number B-10894.

The culture of the strain Pseudomonas putida 131 effectively utilizes phenolic compounds in wastewater, including sulphide-alkaline effluents, under aerobic conditions, using phenolic compounds as a carbon source.

Morphological signs. Rod-shaped cells. Gram-negative.

Cultural signs. On meat-peptone agar (MPA) and selective agar for bacteria of the genus Pseudomonas grows abundantly. MPA has a composition, g / l:

Meat broth 500 g of fresh meat separated from films and fat / liter of water Agar twenty Peptone 10 NaCl 5

On the MPA, it forms round colonies with smooth edges, cream-colored, convex with an eminence in the center, of a uniform consistency, with a diameter of up to 5 mm. On a selective medium for bacteria of the genus Pseudomonas forms cream-colored colonies, shiny, convex, of uniform consistency, with smooth edges and a flat surface, with a diameter of up to 10 mm. The strain grows well in the mineral environment of the composition shown in table 1.

Table 1 The composition of the mineral medium for the cultivation of bacteria Substance Content, g / l KH ₂ PO ₄ 0.16 Na ₂ HPO ₄ 0.55 MgSO ₄ 0.20 (NH ₄ ) ₂ SO ₄ 0.50 FeSO ₄ 0.01 CaCl ₂ 0.01 phenol 0.5 distilled water up to 1 l

In this case, a yellow pigment with a greenish tint is formed.

Physiological signs. Aerob, optimal growth temperature 25 ÷ 30 ° C, lack of destruction of phenolic compounds at 15 ° C.

Biochemical signs. The culture is oxide-positive, utilizes glucose, sorbitol, mannitol, inositol, has catalase and arginine dehydrogenase activity, gives a positive reaction to β-galactosidase, does not utilize citrate, malonate, lactose, sucrose, is not capable of decarboxylation of lysine and ornithine, does not form does not have urease activity. The strain does not form poly-β hydroxybutyrate, is not capable of denitrification.

When culturing in the mineral medium of the composition shown in table 1, on a shaker at a speed of 180 rpm and a temperature of 26 ° C, the Pseudomonas putida 131 strain completely destroys phenol at a concentration of 500 mg / l in 12 hours.

The culture can be stored in a selective medium for bacteria of the genus

Pseudomonas and MPA, at pH 7.0 at 4 ÷ 8 ° C for 3 months.

Cultivation in the laboratory. For the destruction of phenolic compounds, the culture is washed off the surface of the mowed agar medium in a test tube with a small volume of 0.5 ÷ 1.0 ml of sterile tap water or a 0.9% solution of sodium chloride and introduced into 3 ÷ 5 ml of sterile meat and peptone broth of the following composition, g / l:

Meat broth 500 g fresh meat, separated from films and fat / liter of water Peptone 10 Nad 5

Grown for 12 ÷ 18 hours at a temperature of 25 ÷ 30 ° C. The resulting culture infects the mineral medium described above, in which, when cultured in flasks on a rocking chair, the complete destruction of phenol takes place in 12 hours.

The strain belongs to the group of non-virulent, nontoxigenic and nontoxic according to the conclusion of the Ufa Scientific Research Institute of Occupational Medicine and Human Ecology of Rospotrebnadzor.

The method is as follows. The wastewater to be treated is fed into a bioreactor, which is a column-type tank filled with a microorganism strain Pseudomonas putida 131, immobilized onto an oil-calcined coke, biofilter 1. To prevent coke entrainment, a grid is installed in the lower and upper parts of the reactor. When the wastewater contaminated with phenolic compounds passes through the reactor through the reactor, the phenolic compounds are destroyed by a strain of microorganisms uniformly distributed in the reactor volume in the presence of air oxygen supplied through an air distributor 2. To preserve the activity of the microorganism strain, a biogenic additive is introduced into the bioreactor, previously dissolved in wastewater. Gases formed as a result of the vital activity of bacteria are discharged from above.

In a particular case, the wastewater supplied for treatment contains sulfide ions up to 64.1 mg / l. As a biogenic additive, orthophosphoric acid or salts of orthophosphoric acid can be used.

The cells of the microorganism strain are immobilized onto an oil calcined coke based on a heavy pyrolysis resin by a funnel application method, which consists in pumping (with recycling) a suspension of cells through a funnel filled with an adsorbent carrier. The size of the coke fraction is 5 ÷ 10 mm. For immobilization, the adsorbent and cell suspension were used in a ratio of 800 ml of carrier to 3 l of cell suspension. The adsorbent was previously washed with water and sterilized at 1 atm. within 30 minutes The most optimal pH value for the destruction of phenol with an immobilized strain is 7–8; therefore, the sulfide – alkaline runoff was preliminarily neutralized with acid.

The method is illustrated in the drawing, which shows a bioreactor, which is a column type tank, inside of which there are a biofilter 1 and an air distributor 2.

The method is illustrated by examples.

Example 1. In sample No. 1 of sulfide-alkaline runoff of JSC Gazprom Neftekhim Salavat (composition is shown in table 2), pre-neutralized with sulfuric acid to a pH of 7.5, with a concentration of phenolic compounds of 300 mg / l, salt was added as a biogenic additive phosphoric acid K ₂ HPO ₄ at a final concentration of 80 mg / L. Next, the sample was passed through the bioreactor shown in the drawing, filled with a microbial strain of Pseudomonas putida 131 immobilized on an oil calcined coke, while sparging with air at a flow rate of 2 l / h for 4 hours. The results are shown in table 2.

table 2 The composition of sample No. 1 before and after cleaning Indicators Sulfide-alkaline runoff, mg / l Source Purified pH 7.5 8.9 Mass concentration of sulfide ions, mg / l 28.6 8.8 Mass concentration of sulfate ions, mg / l 401.4 189.1 Mass concentration of phenolic compounds, mg / l 300 0.8

Example 2. Sample No. 2 sulfide-alkaline runoff of JSC Gazprom Neftekhim Salavat with a sulfide ion content of 64.1 mg / l (composition is shown in table 3), pre-neutralized with phosphoric acid, with a concentration of phenolic compounds of 800 mg / l, was passed through the bioreactor, as shown in example 1, for 15 hours. The results are shown in table 3. Thus, the invention is shown in the range of content in the wastewater sulfide ions to 64.1 mg / L.

Table 3 The composition of sample No. 2 before and after cleaning Indicators Sulfide-alkaline runoff, mg / l Source Purified pH 7.5 8.8 Mass concentration of sulfide ions, mg / l 64.1 25.6 Mass concentration of sulfate ions, mg / l 1843.7 544.8 Mass concentration of phenolic compounds, mg / l 800 fifty

Example 3. Sample No. 3 of the mineral medium (the composition is shown in table 4), with a concentration of phenolic compounds of 300 mg / l, was passed through a bioreactor, as shown in the previous examples. After 4 hours, the concentration of phenolic compounds is less than 50 mg / L.

Table 4 The composition of the mineral medium containing phenolic compounds Substance Content, mg / L KH ₂ PO ₄ 160 Na ₂ HPO ₄ 550 MgSO ₄ 200 (NH ₄ ) ₂ SO ₄ 500 FeSO ₄ 010 CaCl ₂ 010 Phenolic compounds 300

Example 4. In samples of sulfide-alkaline runoff, previously neutralized with hydrochloric acid to pH 7.5, with a content of phenolic compounds of 500 mg / l, sodium chloride, sulfuric and phosphoric acid salts NaCl, Na ₂ SO ₄ , Na ₂ were added as a biogenic additive HPO ₄ , KH ₂ PO ₄ . One sample was neutralized with phosphoric acid and no salt was added to it. Purification from phenolic compounds was carried out as shown in the previous examples. The results are shown in table 5.

Table 5 The influence of various salts on the activity of the destruction of phenolic compounds using a strain of microorganisms Pseudomonas putida 131 Type and amount of salt, mg / l The content of phenolic compounds, mg / l After 12 hours After 18 h After 24 h NaCl - 4650 354.0 142.2 0.5 Na ₂ SO ₄ - 1650 385.7 324.8 0.4 Na ₂ HPO ₄ - 650 0.3 0.3 0.2 KH ₂ PO ₄ - 650 0.3 0.2 0.1 Sample neutralized with phosphoric acid 100.0 0.3 0.3

Example 5. Sample No. 5 sulfide-alkaline runoff of JSC Gazprom Neftekhim Salavat (composition is shown in table 6) was diluted 2 times in a conical flask with non-sterile tap water to a final volume equal to 30% of the volume of the flask. Then, K ₂ HPO ₄ salt was added at a final concentration of 300 mg / L and 10% by volume of the culture of the Pseudomonas putida 131 strain grown, as described above, on a phenol mineral medium. Then the bacteria were cultured on a rocking chair for 12 h at a temperature of 25 ° C and a rocking speed of 180 rpm. The results are shown in table 6.

Table 6 An example of the destruction of phenolic compounds in sulfide-alkaline stock strain Pseudomonas putida 131 Indicators Sulphide-alkaline stock Source After destruction pH of the medium 7.5 8.9 Mass concentration of sulfide ions, mg / l 29.8 9.6 Mass concentration of phenolic compounds, mg / l 204.6 Lack of

Example 6. Sample No. 6 sulfide-alkaline runoff of JSC Gazprom Neftekhim Salavat, containing phenolic compounds at a concentration of 1200 mg / l, was diluted twice with tap water and the pH was adjusted to 7.5 with concentrated sulfuric acid. Then, Na ₂ SO ₄ salt was introduced so that its final concentration was 4650 mg / L (in terms of sulfate ion 3143 mg / L), and the total mineralization was 5400 mg / L. Bacteria of the strain Pseudomonas putida 131 were cultivated on a rocking chair, as described in example 5. The results are shown in table 7.

Table 7 The destruction of phenolic compounds by bacteria Pseudomonas putida 131 in a sulfide-alkaline effluent with a high content of sulfate ions Option The content of phenolic compounds, mg / l, through 12 hours 18 hours 24 hours Pseudomonas putida 131 459.0 437.8 0.6 Bacteria free 598.7 563.9 524.8

Example 7. For an example of the destruction of phenolic compounds, the mineral medium of the composition shown in Table 8 was used. The bacteria of the strain Pseudomonas putida 131 were cultured on a rocking chair, as described in Example 5. After 40 hours of cultivation on a mineral medium, phenol compounds were not determined.

Table 8 The composition of the mineral medium containing phenolic compounds Substance Content, mg / L KH ₂ PO ₄ 160 Na ₂ HPO ₄ 550 MgSO ₄ 200 (NH ₄ ) ₂ SO ₄ 500 FeSO ₄ 010 CaCl ₂ 010 Phenol 352 P-cresol 029 O-cresol 017 Distilled water up to 1 l

Effective destruction of phenolic compounds in various media suggests the possibility of rapid adaptation of the strain to changing environmental conditions, i.e. confirms the high adaptive ability of the strain Pseudomonas putida 131.

The content of phenolic compounds was determined by high performance liquid chromatography.

Thus, the use of the strain Pseudomonas putida 131 provides an increase in the quality of wastewater treatment from phenolic compounds. Sewage treated with a strain can be used for further treatment with domestic wastewater.

Claims (5)

1. The method of purification of wastewater from phenolic compounds using a strain of aerobic bacteria and biogenic additives, characterized in that the strain Pseudomonas putida 131 VKPM B-10894 is used as a strain of aerobic bacteria. 2. A method of treating wastewater from phenolic compounds according to claim 1, characterized in that the wastewater contains sulfide ions up to 64.1 mg / L. 3. The method of treating wastewater from phenolic compounds according to claim 1, characterized in that the wastewater contains sulfate ions in an amount of 401.4 or 1843.7 mg / L. 4. The method of wastewater treatment from phenolic compounds according to claim 1, characterized in that orthophosphoric acid is used as a biogenic additive. 5. The method of wastewater treatment from phenolic compounds according to claim 1, characterized in that salts of phosphoric acid are used as a biogenic additive.

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