SU1752727A1 - Strain of bacteria brivibacterium sp, used for purification of sewage from acrylamide and acrylic acid - Google Patents

Abstract

The use of wastewater treatment from acripamide and acrylic acid. Essence of the strain isolated from the soil by the production of acrylonitrile using propion midz as a growth substrate. l for 28 h with a microbial load of 1 32 g / l of substrate 2 table

Description

This invention relates to applied microbiology and relates to the preparation of a new bacterial strain used for the local treatment of wastewater from acrylamide and acrylic acid.

Acrylmmmd is the basis for the production of polymeric materials with a wide range of applications. Polymers and copolymers of acrylamyl are used in the chemical petroleum and mining industries in the pulp and paper industry. Acrylamide and cellulose products can be used as thickeners in dyes for fabrics. Acrylamide can be used for specific chemical modification of proteins which is important for studying the mechanism of catalytic action of enzymes

Modern technology and production of acripamide does not prevent the plant from being released into the atmosphere and wastewater (insufficient tightness of reactors; open surfaces on filters; low efficiency of ventilation systems; open viewing hatches of reactors). For additional wastewater treatment from akoylsmide, microbiological methods are most effective and cost-effective. detoxification methods

Known strains analogs capable of degradation of acrylamide and acrylic acid

Closest to the proposed strain is the culture of the bacterium Brevibacterium ammoniaqenes I AM 1641, which utilizes for 48 hours 1 g / l acrylamide with microbial load of 1 g / l (raw weight) The same culture destroys 0.2 g / l acrylamide 96% contained in wastewater in 24 hours

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The aim of the invention is to obtain a strain with high biodestructive activity capable of degrading acrylamide and acrylic acid in wastewater at higher concentrations and with greater speed.

The resulting strain Brevibacterium sp. 12 PAs are deposited at the Central Museum of Industrial Microorganisms, All-Union Scientific Research Institute of Genetics, and are characterized by the following morphological, cultural, and physiological properties.

Morphological properties. Young cells of the strain are in the form of medium sized sticks, arranged singly, in pairs and in small chains. Grampositive. Aging culture (2-3 days) is represented by coccobacilli located randomly. Spores and capsules do not form. Mobile, The strain multiplies by simple division, forming short, unbranched sticks.

Cultural properties. Prototroph, aerobic, optimal growth temperature 28 - JSPg :. It grows at 5 - 10 and 40 ° C. The optimum pH for culture development is 7.4 ± 0.2, although growth is also observed when the pH of the medium is 6–9 Colonms on medium-sized peptone agar (2–3 mm), pigmented, and the co-yellow pigment. During cultivation, the strain on special media does not form water-soluble or fluorescent pigments. Colonies of the correct rounded shape, opaque. The rim around the colonies is flat, the colonies are slightly convex. The edge of the colony is smooth, the surface is smooth, glossy. On moc-peptone broth, the growth is in the form of a uniform turbidity of the whole medium and a flocculent sediment on the bottom.

Biochemical properties, Oxydazone-ngative, catalazopositive, Gelatinase, Phenylellandeaminase, lipase, lecithin, arginine hydralase, lysine decarboxylase, ornithine decarboxylase, does not form urease. Tests on the OF medium with glucose, maltose, fructose give a positive reaction, on the medium with arabinose, xylose, mannitol - alkalization, On media containing lactose, sucrose, rhamnose, mainoses, miliosis, raffinose, trehalose, sorbitol, inositol, dulcite, adonit, salicin, inulin - alkalinity. It grows in the presence of 6.5% NaCl, reduces nitrates to nitrites. Does not grow on Mac-Conkey and Ploskarev. On red blood cell hemolysis does not give erythrocytes. Indole and hydrogen sulfide does not form.

The pathogenicity of the strain was determined in white mice by intraperitoneal administration.

by the suspension of bacteria 5,109 cells / ml. The absence of any pathological changes in the internal organs of mice suggests that culture is avirulent.

The strain Brevibacterium sp 13 PA was isolated from the soil from the production of acrylonitrile by the method of cumulative culture using as growth substrate

propionamide.

The essence of the invention is that the proposed strain is capable of destroying 4 g / l acrylamide and 2.5 g / l acrylic acid in a mineral medium for 24 hours (microbial load of 10 cells / ml), using them as the sole source of carbon and nitrogen in case of acrylamide or as a carbon and energy source in case of acrylic acid.

PRI m s p 1. Isolation of strain

Brevibacierium sp.13 PA.

The microorganism-destructor of acrylamide and acrylic whispers isolated from the soil from the production of acrylonitrile by the method of cumulative culture. A weighed sample of 1 g was carefully resuspended in 10 ml of physiological solution and infused for 1 h. The 1 ml supernatant was added in the following composition, g: glucose 0.5; peptone 1.0; CanRO 0.1; 0.1; distilled water 1000 ml; pH 7.0 iO, 2; propionamide - initial concentration 0.1 g / l

Suspension (50 ml) in Erlenmeyer

300 ml flasks are incubated at 28–30 (° C with circular agitation. Daily the culture liquid was half-renewed. The concentration of propionamide in the medium was increased step by step from 0 1 to 20 g / l.

During the month of adaptation, peptone and glucose were completely removed from the medium. After two months of passaging on propionamide, the culture liquid was sown on plates with a dense nutrient medium, g / l: MgSO / i 0.1; CgNRO 0.1, gpitserin 5; acrylamide 5; ai ar-agar 1.5; pH 7.0 ± 0 2.

As a result, the culture of Brevibacterium sp. 13 PA, recycling

acrylamide, acrylic acid as the sole source of carbon and nitrogen, or carbon and energy. The destructive activity of the strain was tested in a liquid synthetic medium,

Acrylamide and acrylic acid were quantitatively determined by liquid chromatography on an LCM-80 chromatograph, with a flame ionization detector. Roop ex-400 was used as the stationary phase. Time

analysis of acrylamide 8 min, acrylic acid 3 min.

PRI mme R 2. The use of a strain of Brevibacterium sp 13 PA as a destructor of acrylamide.

The strain, previously induced during 24 hours on a nutrient medium with propionamide as an inductor (5 g / l), was cultured in Erlenmeyer flasks on a liquid synthetic medium of composition, g / l, MgSCM 0.2; K2HP04 0.1; distilled water 1000 ml; pH 7.2 - 0.2, in a volume of 50 ml. Microbial load of 10, 10, 10 cells / mp. As a source of carbon and nitrogen, ecrylamide was introduced at a concentration of 2.5 and 5.0 g / l. The flasks were placed on a shaker (120 v / mich) and incubated at 28-30 ° C. The acrylamide concentration was determined by GLC.

PRI me R 3 Use of a strain of Brevibacterium sp 13 PA as a destructor of acrylic acid.

The strain was cultivated as in Example 2. Acrylic acid at a concentration of 1 and 2.5 g / l was adjusted to pH 7.0 with a 10% aqueous solution of ammonia, which also serves as a source of nitrogen. Microbial load of 10 cells / ml. No prior induction of the strain was required in the event of destruction of acrylic acid.

Example 4 Test of Brevibacterium sp, 13 PA for wastewater treatment.

Wastewater from the pilot production of acrylamide and liaacrylamide was inoculated with a culture of Brevibacterium sp. 13 PA (microbial load 10 cells / ml). Exposure time 24 hours, temperature 28 ° C, occasional stirring. The bacterial mass was separated by centrifugation for 40 minutes at 7000 rpm. The concentration of acrylamide and acrylic acid was determined as in Example 1.

The results of testing the biodestructive activity of the proposed strain, as well as the known, are given in Tables 1 and 2.

The proposed strain significantly exceeds the known for its destructive characteristics (the rate of substrate utilization and the maximum utilized substrate concentrations) The tested culture destroys 5 g / l of acrylamide in 28 hours. The presence of acrylic acid in the medium does not affect the destructive activity of the proposed strain with respect to acrylamide. After complete disappearance from the environment, acrylamide begins

0 the process of utilization of acrylic acid, formed equivalent to the quantities of destroyed acrylamide. In addition to otogg, the strain can utilize akrmlov,., Acid as an independent single carbon source with its maximum content in an environment of 2.5 g / l in 24 hours.

Testing of the strain in real industrial effluent showed its viability and high enzymatic

0 activity, It is established that in 24 hours the culture of Brevibacterium sp. 13 PA destroys 4.37 g / l of acrylamide contained in wastewater. Acrylic acid acts as a metabolite of the process.

5 is further utilized by the culture in 24 hours, protrusion as a source of carbon and energy.

Tests conducted on 5 samples of wastewater. The presence of acrylonitrile 0.43% in the effluents, zinc salts (11) 1.82 mg / l, copper (II) 10.0 mg / l, iron (III), serbital and toluene does not reduce the biodestructive activity of the strain.

The destructive activity of the strain is carried out in a wide range of pH 6-9, therefore the need to regulate the pH of the effluent is eliminated.

Thus, the proposed strain Brevibacterium sp. 13 PA has significant destructive activity, has been tested in industrial wastewater and can be recommended for use for local wastewater treatment from acrylamide and acrylic acid.

Claims (1)

5 claims The strain of bacteria Brevibacterium sp. VKPM B-4987 used to treat acrylamide and acrylic acid from wastewater. T g b l and c a 1 Strain Initial substrate concentration i / l Brevlbacterium sp 13 PA B -4987 Brevibacterium dmmoniagenes AM 1641 (known) Microbial load 1 QЈ-cy2ЈMyj5Cii.Jl / iL T eblitsa 2 Disposal time, h 1.32 28 48