JPH1033163A - Baterium capable of decomposing malodorous component - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a bacterium of the genus Pseudomonas capable of efficiently decomposing malodorous components contained in an aqueous waste liquor discharged from a coating plant, etc., especially malodorous alcohols, ketones, ethers, esters and organic acids. SOLUTION: A Pseudomonas N-TMS-5 species (FERM-P-15525) having the ability to efficiently decompose malodorous components is obtained. The strain is capable of surviving even in a saturated aqueous liquor of an aromatic solvent used in a large amount as a coating solvent, e.g. benzene, xylene or toluene and treating a waste liquor containing this kind of coating solvent.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a Pseudomonas which efficiently decomposes malodorous components contained in aqueous effluent discharged from a coating plant and the like, in particular, malodorous alcohols, ketones, ethers, esters and organic acids. A new bacterium of the genus.

[0002]

2. Description of the Related Art In an environment in which various organic solvents are mixed, such as a coating booth pit tank of a coating plant, malodorous components such as malodorous alcohols, ketones, and ethers due to the action of putrefactive bacteria. , Esters, organic acids and the like are generated, which is a serious problem not only in the working environment in the factory but also in the living environment around the factory. For this reason, in the painting booth pit tanks, etc., liquid replacement is performed periodically as a countermeasure against odor, but a huge amount of replacement water of several hundred tons is required, which is extremely impractical in terms of cost and time. In addition to being economical, there is a problem in that a large amount of aqueous wastewater containing malodorous components must be treated.

[0003]

SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned circumstances in the art, and has been made to provide an effective measure for treating an aqueous waste liquid containing a malodorous component.

[0004]

That is, the present invention relates to Pseudomonas N-TMS-5 (Ps
eudomonas sp. N-TMS-5; FERM-P-155
25) and a method for treating malodorous component-containing aqueous waste liquid using the bacteria. In the present specification, the "odorous component" emits an offensive odor among various organic solvents generally used in the field of coatings, for example, alcohols, ethers, esters, ketones, and organic acids. And odorous compounds formed from these organic solvents by the action of spoilage bacteria.

The bacterium according to the present invention is located in Kosai town, Shiga prefecture.
(Wastewater treatment plant in the NEOS Shiga Factory) Separated from activated sludge by the following method. An appropriate amount of the activated sludge is heated under pressure (temperature: 120 ° C., pressure: 1 kgf / cm
² ) Sterilized for 15 minutes in an inorganic salt medium of Stanyl et al. Having the following composition [RY Stanier, NJ Palleroni and M. Doudoro]
ff), J. Gen. Microbiol. 43, p. 159 (1
966)], butanol was added thereto at 1000 pp.
After the addition of the medium, the medium is placed in a culture flask, and 100 r
The culture was subjected to a reciprocal shaking culture treatment at pm for 10 days. Then, the cultured cells were subjected to a subculture treatment using a similar medium several times.
Then, under pressure and heat treatment (temperature: 120 ° C., pressure: 1 kgf /
The above cultured bacterial cell liquid was applied to a standard agar medium sterilized at ² cm ² ), and bacterial colonies that grew predominantly in 1 to 10 days were separated.

Constituent concentration of inorganic salt medium of Stanyl et al. (Mg / 1000 ml) NaHPO ₄ + KH ₂ PO ₄ (1M; pH 6.8) 40 ml (NH ₄ ) ₂ SO ₄ 1000 Nitrilotriacetic acid 200 MgSO ₄ .2H ₂ O 289 _{CaCl 2 · 2H 2 O 66.7 (} NH 4) 6 M o7 O 24 · 4H 2 O 0.185 FeSO 4 · 7H 2 O 1.98 EDTA 2.5 ZnSO 4 · 7H 2 O 10.95 FeSO 4 · 7H ₂ O 5 MnSO ₄ .H ₂ O 1.54 CuSO ₄ .5H ₂ O 0.392 Co (NO ₃ ) ₂ .6H ₂ O 0.248 Na ₂ B ₄ O ₇ .10H ₂ O 0.177

[0007] The mycological properties of the bacteria obtained by the above-mentioned separation method are described in "Classification and identification of microorganisms (2)" [edited by Takeharu Hasegawa,
Published by Gakkai Shuppan Center (1985, revised edition)], No. 99
When examined according to the method described on pages 161 to 161,
It was as follows. Morphological properties a. Shape and size of cells Short rods or rods having a size of 0.5 to 0.7 × 1.0 to 2.0 μm. Filamentous (10-30 μm) in late culture b. Motility flagellar movement c. Gram staining Gram negative d. No sporulation Physiological properties a. Oxygen demand aerobic b. Oxidase positive c. Catalase positive d. OF test oxidative e. Formation of dye Water-soluble fluorescence (light green, King B medium) f. Optimal growth temperature 25 ° C g. Growth pH range 5.0 to 10.0 h. Negative for both growth at 4 and 41 ° C i. Starch degradation positive j. Gelatin degradation negative k. Poly-β-hydroxybutyrate degradation negative l. Arginine degradability positive m. Levan formation negative n. Egg yolk test negative o. Indole production negative p. Hydrogen sulfide production negative q. Nitrate reducing ability negative r. Nitrate respiration negative s. G + C mol% 72.3%

The taxonomic position of the bacterium according to the present invention is based on the above-mentioned mycological properties, and is based on the "Birzy Manual of Deterministic Bacteriology" (8th edition, 1st edition).
974), the bacterium was found to be a novel strain belonging to the genus Pseudomonas. The present inventor has designated this bacterium as Pseudomonas NT
MS-5 species (Pseudomonas sp. N-TMS-5), and the PERM-P
-15525. The present invention includes all mutants obtained by mutating the above-mentioned strains naturally or by artificial means, which have the capability of degrading odor components.

[0009]

BEST MODE FOR CARRYING OUT THE INVENTION The method for culturing the above-mentioned strains according to the present invention is not particularly limited, and a medium generally used for culturing strains in the art may be appropriately selected and carried out. The method comprises the use of the above-mentioned Stanyl et al. Inorganic salt medium as a carbon source as a malodorous component such as butanol or a lower alcohol or polyhydric alcohol such as methanol or ethanol (for example, ethylene glycol,
Propylene glycol, methyl cellosolve and ethyl cellosolve) in an amount of 10 to 10,000 ppm, preferably 1 to 10 ppm.
This is a method using a medium added at a concentration of 00 to 1000 ppm. A preferred culture method is a liquid reciprocal shaking culture,
The culture temperature is 15 to 30 ° C., preferably 23 to 27 ° C., and the pH of the medium is 5 to 5 using a sodium hydroxide solution or the like.
It is generally adjusted to 9, preferably 6.5 to 7.5.

The bacterial strain according to the present invention can survive even in a saturated aqueous solution of an aromatic solvent (for example, benzene, xylene, toluene, etc.) which is used in large quantities as a coating solvent. It can be used without problem for the treatment of the aqueous effluent containing the malodorous component.

A general method for treating an aqueous effluent containing a malodorous component using the strain according to the present invention is as follows: a culture solution containing the strain cultivated as described above or a powder obtained by freeze-drying the strain is treated with the aqueous solution to be treated. This is a method of adding to the waste solution and performing aeration.The strain is immobilized on a suitable carrier (for example, activated carbon, porous silica, resin, κ-carrageenan, calcium alginate, etc.), and the carrier is treated with the aqueous solution. Waste liquid and air may be supplied. The treatment of aqueous odor containing odorous components
Usually, under aerobic conditions at a pH of 5 to 9 (preferably 6.5 to 7.5) and a temperature of 15 to 30 ° C (preferably 23 to 27 ° C), the malodorous components are sufficiently assimilated. Until it is done. In this case, an inorganic nitrogen source (for example, ammonium nitrate and the like), a phosphate source (for example, potassium phosphate and the like), inorganic salts (for example, magnesium salt, iron salt and the like) and / or organic growth factors (for example, yeast extract, peptone and the like) May be appropriately added to the aqueous waste liquid to be treated to promote the growth of the bacterial strain and promote the decomposition of the malodorous components. The amount of the strain used depends mainly on the concentration of the malodorous components in the aqueous wastewater to be treated,
Although not particularly limited, for example, when the concentration of the
1 × 10 ³ -1 ×
It is 10 ⁹ cells / ml, preferably 1 × 10 ⁴ to 1 × 10 ⁶ cells / ml. Under the above conditions, the malodorous component in the aqueous waste liquid to be treated can be completely decomposed in 1 to 7 days.

The present invention is mainly directed to aqueous waste liquids such as a coating booth pit tank of a coating plant, but is also applicable to an activated sludge treatment of an aqueous liquid containing malodorous components. In addition, components that inhibit the treatment or reduce the efficiency of the treatment (for example, a preservative, a rust inhibitor, an antibacterial agent, and the like) may be appropriately removed before performing the microorganism treatment.

[0013]

The present invention will be described below with reference to examples. Example 1 An Erlenmeyer flask (100 ml) was charged with Stanyl's inorganic salt medium (20 ml), stoppered with foamed silicone rubber,
This was subjected to a sterilization treatment under pressure and heating (120 ° C .: 1 kgf / cm ² ). After sterilization, the flask was opened under sterile conditions, and any of the following malodorous components (1000 ppm) was added to the medium as the sole carbon source, and Pseudomonas N-TMS-5 species cultured as described above. Was inoculated (approximately 1 × 10 ⁴ cells / ml), a foamed silicone rubber stopper was attached to the flask, and this was subjected to shaking culture treatment at 25 ° C. for 4 days. Alcohols: propanol, n-butanol, iso-butanol, n-pentanol ketones: diisopropyl ketone, diisobutyl ketone ethers: di-n-butyl ether esters・ Butyl acetate Organic acids ・ ・ ・ ・ ・ Acetic acid, propionic acid, butyric acid, formic acid When the analysis of the malodorous component in the culture solution was performed by gas chromatography under the following conditions, the malodorous component was not detected. No odor was found. In addition,
The effect of decomposing and eliminating formic acid was measured at a wavelength of 660 using a spectrophotometer.
This was confirmed by a significant increase in absorbance at nm (> 0.1). Column PEG-20M 3mmφ × 1m manufactured by Nippon Chromatography Co., Ltd. Liquid phase: Polyethylene glycol 20M Carrier: Chromosorb W (HP) Impregnation rate: 10% Mobile phase Nitrogen gas (40 ml / min) Initial column temperature 50 ° C Final column temperature 170 ° C rise Temperature condition 6 ℃ / min Injection temperature 180 ℃ Detector temperature 180 ℃ Detector Flame ionization detector (FID) Injection volume 2μl

Comparative Example 1 The same treatment as in Example 1 was carried out except that no strain was inoculated. As a result, no decomposing / eliminating effect was found on any offensive odor components.

Example 2 About 200 ppm of butyric acid as a malodorous component obtained by filtering solids such as paint residue from an aqueous waste liquid collected from a coating booth pit tank.
Was inoculated with a strain according to the present invention (about 1 × 10 ⁴ cells / ml) and shaken at 25 ° C. When the treatment liquid after the above treatment was performed for 4 days was analyzed by gas chromatography described in Example 1, butyric acid was not detected and no malodor was recognized.

Example 3 0.1% (v / v) of methanol, ethanol, ethylene glycol, propylene glycol, methyl cellosolve or ethyl cellosolve as a carbon source instead of malodorous components.
The culture treatment was performed in the same manner as in Example 1 except for adding. The increase in turbidity of the culture with the growth of the strain was measured using a spectrophotometer. In each case, the absorbance at 660 nm showed a remarkable increase of 0.1 or more.

Example 4 Toluene or xylene was added at 0.1% (v / v) after inoculation of the strain.
v) Culture treatment was carried out for 7 days in the same manner as in Example 1 except for adding. When the culture solution was analyzed by gas chromatography described in Example 1, no malodorous components were detected, and no malodor was recognized.

Example 5 200 ml of an inorganic salt medium of Stanyl et al.
And porous sintered glass filter media (Siporax
35 g (made by SCHOTT GERMANY CO., LTD.) Was put in the container, and the container was plugged with foamed silicone rubber.
^It was subjected to the sterilization treatment in ² ). After sterilization, the flask was opened aseptically and 0.1% (V / V) was added to the medium as a carbon source.
Butanol was added, and Pseudomonas N-TMS-5 strains cultured as described above were inoculated.
(About 1 × 10 ⁴ cells / ml), a foamed silicone rubber stopper was attached to the flask, and this was subjected to a shaking culture treatment at 25 ° C. for 7 days. After the cultivation, the filter medium to which the strain was adhered was removed to an inner diameter of 3.5 c.
m, 13 cm long, packed in a 100 ml column,
The plate was washed sufficiently with a sterilized inorganic salt medium of Stanyl et al. 300 ml of the odor sample described in Example 2 was circulated at a flow rate of 10 ml / min through the thus-formed cell-immobilized column, and the column was ventilated at 0.51 / min. Was performed. When the treated liquid after one day of the above treatment was analyzed by gas chromatography described in Example 1, butyric acid was not detected and no bad odor was recognized.

Comparative Example 2 The same treatment as in Example 5 was carried out except that the strain was not inoculated. However, even after 10 days, butyric acid was not lost, and an odor remained.

[0020]

EFFECT OF THE INVENTION By utilizing the bacteria of the genus Pseudomonas according to the present invention, malodor components, especially malodor components such as hydrocarbons, alcohols, ketones, ethers, esters and organic acids are contained. Aqueous effluents can be efficiently treated at low cost in a relatively short period of time, greatly improving the working environment in factories and the living environment around the factories, which has been a problem for waste odors. it can. The present invention is mainly directed to an aqueous waste liquid discharged from a coating plant or the like, but is also applicable to an activated sludge treatment of an aqueous waste liquid containing a malodorous component.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁶ Identification number Agency reference number FI Technical indication location // (C12N 1/20 C12R 1:38) (72) Inventor Masaaki Yamamoto Nakamura-ku, Nagoya-shi, Aichi 3-25-9 Meinoeki Horiuchi Building Neos Nagoya Sales Office

Claims (4)

[Claims] 1. Pseudomonas sp. N-TMS-5 having an odor component resolution.
FERM-P-15525). 2. The bacterium according to claim 1, wherein the malodorous components are malodorous alcohols, ketones, ethers, esters and organic acids. 3. An aqueous waste liquid containing an offensive odor component.
Or a method for treating the aqueous waste liquid, the method comprising treating with the bacterium according to 2. 4. The method according to claim 3, wherein the malodorous component is malodorous alcohols, ketones, ethers, esters and / or organic acids.