RU2041944C1 - Strain of bacteria paracoccusdenitrificans being destructor of aniline, 3-chloraniline,4-chloraniline and 3,4-dichloraniline - Google Patents

Abstract

FIELD: biotechnology. SUBSTANCE: is prepared strain Paracoccus denitrificans. Said strain is destructor of aniline, 3-chloraniline, 4-chloraniline and 3,4-dichloraniline. EFFECT: improves quality of desired product.

Description

 The invention relates to the microbiological industry and relates to a new strain of the destructor 3,4-dichloroaniline (DCA), 3-chloraniline (ZXA), 4-chloraniline (4XA), promising in the work on wastewater treatment from pollution.

A known strain of Pseudomonas putida, which in the presence of aniline mineralizes DHA / 5 mg / l for 40 hours [1]
The closest in technical essence and the achieved positive effect is the strain Pseudomonas diminuta INMI KS-7VKM-148g, using DCA as the sole source of carbon, nitrogen and energy. The maximum concentration at which there is an increase of 50 mg / l. This amount of DCA under aeration disappears in 96 hours. A characteristic feature of the strain is the absolute need for meteonin as a growth factor [2]
The disadvantages of the known strains are a relatively low level of destructive activity, inhibition of microorganisms with an increase in the concentration of DCA in the culture medium above 50 mg / L.

 The disadvantage of the strain P. diminuta INMI KS-7 VKM-148g, in addition to these qualities, is the need to introduce methionine into the culture medium, which under industrial conditions leads to an increase in the cost of the cleaning process.

 The purpose of the invention to obtain a strain of the destructor of aniline and chloralin with high activity and simple nutritional needs.

 The bacterial strain was obtained by isolating the fastest growing colony on an agarized synthetic medium with DCA after prolonged exposure to soil taken from Kuban draftsmen in a medium containing DCA.

 The resulting bacterial strain was deposited in the All-Union Collection of Industrial Microorganisms (VKPM) under the number S-1193.

 The strain P.denitrificans VKPM-S-1193 is characterized by the following cultural-morphological and physiological-biochemical characteristics.

 Spherical cells of 0.5-1 μm in diameter are found in pairs and individually. Fixed, gram-negative. Meat peptone agar (28, 48 h). It forms shiny, mucous, round, white colonies with a diameter of 2-4 mm. When stored in a refrigerator on solid agarized media, mucosal colony increases. Mineral environment with DHA. The colonies are round with smooth edges, shiny, mucous with a diameter of 1-2 mm. Liquid mineral medium with DHA. Forms turbidity, a dense precipitate that easily agitates when shaken. The pigment does not emit on Wednesday.

Aerob, gives a positive catalysis and oxidase reaction, accumulates polyoxybutyric acid in the cells. It grows at 36 ^{° C,} does not grow at 4 ° ^C. The optimum temperature for growth 28-30 ° ^C Optimum pH 6.7-7.2. Starch and gelatin do not hydrolyze. The nitrate is reduced to N. Calculation anaerobically with nitrite as an electron acceptor.

 It is well absorbed and grows on D-glucose, D-fructose, sucrose, trehalose, acetate, propionate, lactate, pyruvate, citrate, succinate, n-hydroxybenzoic acid, benzoic acid.

 It is well stored in tap water throughout the year.

 The strain was identified by the Bergi Identifier (19) as a strain of the species Paracoccus denitrificans.

PRI me R 1. Inoculum is obtained by growing a strain taken from agarized synthetic medium with 4XA. The composition of the medium, g / l: KHPO 7.0; KHPO 3.0; MgSO ₄ 0.1; DHA 0.1-0.12; distilled water.

Cultivation is carried out at 30 ^about C for 48 hours

The prepared seed in the amount of 10 cells / ml is added to 750 ml flasks containing 100 ml of medium of the same composition. Under these conditions, DHA disappears in 96 hours; the release of chlorine ion is 100%
PRI me R 2. Inoculum obtained by the method described in example 1, contribute to 750 ml flasks containing 100 ml of synthetic medium, where DCA is replaced by 4XA 0.5-0.7 g / L. Under these conditions, 4XA disappears in 24 hours, the release of chlorine ion is 100%
PRI me R 3. Inoculum obtained by the method described in example 1, contribute to 750 ml flasks containing 100 ml of synthetic medium, where aniline 5 g / l was taken as a growth substrate. Under these conditions, aniline disappears in 48-60 hours.

 Advantages of P. denitrificans HRH strain compared to P. diminuta INMI KS-7 strain: ability to destroy higher concentrations of DCA (up to 130 mg / l), 4XA and 3XA (600 mg / l), use of aniline as a carbon and nitrogen source in high concentrations (up to 5 g / l), which will allow the proposed strain to be used for the treatment of industrial effluents containing aniline, the absence of dependence on growth factors, which allows saving costs for the cleaning process under production conditions.

 Thus, the strain P. denitrificans KRZ not only surpasses the ability to destroy known strains, but is more technologically advanced in its properties. The claims of the strain Paracoccus denitrificans aniline, 3-chloraniline, 4-chloraniline and 3,4-aniline destructor.

Claims (1)

 The strain of Paracoccus denitrificans VKPMs-1993 is aniline, 3-chloraniline, 4-chloraniline and 3,4-dichloroaniline destructor.