JP2000093164A - New microorganism and dye decoloration using the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a microorganism belonging to genus bacillus having dye decoloring ability under an alkaline condition capable of decreasing the degree of pigmentation of a solution colored by dye, especially useful for processing or the like of dye wastewater used under high temperature and high alkaline condition. SOLUTION: This microorganism [e.g.; bacillus-sp. DA1 (deposited in National Institute of Bioscience and Human Technology as 16921) or the like] belongs to genus bacillus, has decoloring ability to dye at >=40 deg.C under alkaline condition, capable of reducing the degree of pigmentation of a solution colored by dye at middle to high temperature under alkaline condition, especially brings a remarkable effects on the processing of the dye wastewater used under high temperature and high alkaline condition. This new microorganism is obtained by adding and culturing samples of nationwide soils, compost, activated sludge or the like, in an agar medium including Reactive back 5 as an azo reactive dye, at 45 deg.C, separating microorganism forming an orange clear zone in the blue medium after 1 to 2 days, and repeating refinements.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a novel dye decolorizing microorganism and a method for decolorizing a dye using the microorganism.

[0002]

2. Description of the Related Art As a method for decolorizing wastewater colored by dyeing or the like, many studies have been made on physical and chemical decolorization methods such as coagulation sedimentation, ozone oxidation, activated carbon adsorption, hypochlorous acid oxidation, and electrolytic oxidation. Although the degree of coloration of wastewater has been regarded as a problem, it has been receiving special attention since the Wakayama City enacted a color removal ordinance in October 1991. This ordinance came into effect in April 1994, but none of the above methods was a technology that could be handled in terms of processing costs, and the regulation values were relaxed in July of the same year. Since then, regulations have not expanded beyond Wakayama City, but it is thought that regulations will expand at a stretch if technologies that have processing costs, space, and stability that exceed practical levels are developed.

[0003] In the biological method, the decolorization reaction generally proceeds under mild conditions, and there are few secondary problems as compared with the physicochemical method.
Also, since it is considered to be advantageous in terms of cost, active research has been made so far, and many microorganisms having decolorizing ability have been provided. Examples include decomposition of basic dyes by Pseudomonas spp. Bacteria (No. 1283), and benzene ring-containing dyes by Nocardia spp. Bacteria (ATCC 19070, 19071). Degradation (JP-A-48-56881), a bacterium belonging to the genus Alcaligenes (No.
Decolorization of monoazo, diazo, triphenylmethane, methine and monoazo polymer dyes under alkaline conditions (JP-A-63-216472).
Degradation and decolorization of phthalocyanine by filamentous fungi belonging to the genus Myrocesium (No. 10728).
No. 8), Bacillus sp.
No. 13), Xanthomonas sp.
No. 13), Achromobacter sp.
No.) Color elimination or reduction of azo dye coloring liquid by each bacterium (Japanese Patent Application Laid-Open No. 8-261), decomposition and decolorization of dye by a fungus of the genus Geotricum (No. 15348). No. 173051) and the like.

On the other hand, there are various types of dyes, which are used in accordance with the material to be dyed. However, those used in large amounts include disperse dyes mainly used for dyeing polyester and dyes for cotton and the like. The reactive dyes used are mentioned. In particular, reactive dyes are water-soluble and pose a problem as a coloring component of wastewater. Further, from the basic skeleton of the chemical structure of the dye, there are mainly an azo type and an anthraquinone type, and the azo type dye is used more frequently. Therefore, the most problematic dye for coloring wastewater can be called an azo-based reactive dye.

[0005] Dyeing with a reactive dye varies depending on the type of dye.
10-11). Therefore, wastewater after dyeing is completed
Although the temperature drops slightly, it is a medium to high temperature and high alkali. If such wastewater can be directly treated, neutralization and cooling are not required, and cost reduction and efficiency can be expected. However, microorganisms that can decolorize under this condition have not been known so far.

[0006]

SUMMARY OF THE INVENTION An object of the present invention is to provide a method for efficiently decolorizing a dye by a microorganism. Furthermore, microorganisms that can decolor azo-based reactive dyes, which are the most problematic as a coloring source of wastewater, under high temperature and high alkali conditions immediately after dyeing, and an efficient decolorization method using this Is to provide.

[0007]

SUMMARY OF THE INVENTION The present invention relates to a microorganism belonging to the genus Bacillus and capable of decolorizing a dye under alkaline conditions and / or a temperature of 40 ° C. or higher, and a method for decolorizing a dye using the microorganism.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION The microorganism of the present invention belongs to the genus Bacillus and has the ability to decolorize a dye under alkaline conditions (particularly pH 9 or more) and / or a temperature of 40 ° C. or more.

[0009] The microorganism of the present invention was obtained as follows. Samples of soil, compost, activated sludge, etc. from various parts of the country were analyzed using reactive black 5, which is an azo-based reactive dye, at 0.025%, peptone at 0.5%, and yeast extract at 0.1%.
An agar medium (pH 10.5) containing 2%, 0.1% dipotassium phosphate, 0.02% magnesium sulfate heptahydrate, and 1% sodium carbonate was added and cultured at 45 ° C. After 1-2 days, microorganisms forming an orange clear zone in a blue medium are separated, and purification is repeated.
Bacterial strains of the species were isolated. This strain is 30 times in the same medium as above.
When cultured at 30 ° C or 45 ° C in a medium (pH 7.4) without the addition of sodium carbonate, no clear zone was formed even after 3 days.

The bacteriological properties of the present strain are shown below. (a) Morphological properties (1) Cell shape and size Bacillus (0.8 x 4 μm) (2) Motility (flagellate) Yes (perioflagellate) (3) Spore Yes (spherical) (4) Spore sac One end is bulging

(B) Cultural properties (tested at pH 10.3) (1) Broth agar plate culture The colonies are medium in size, convex, smooth in surface, jagged at the periphery, yellowish white, opaque (2) Broth liquid culture Slight turbidity (3) Broth gelatin puncture culture Liquefaction

(C) Physiological properties (1) Gram stainability + (2) Reduction of nitrate + (3) Denitrification reaction-(4) VP test + (5) Formation of indole-(6) Formation of hydrogen sulfide − (7) Hydrolysis of starch + (8) Hydrolysis of casein + (9) Use of citric acid + (both Simmons and Christensen media) (10) Inorganic nitrogen source + (both ammonia and nitrate) (11) Formation of pigment-(12) Urease + (13) Oxidase + (weak) (14) Catalase + (15) Growth range Growth at 25 ° C to 60 ° C (optimum temperature is 45 ° C) pH 7.5 to 12. Grow at 0 (optimal pH is 9 to 10) (16) Effect of NaCl Grow in the presence of 10% (17) Attitude to oxygen Facultative anaerobic (18) O / F test O / F (19) L-arabinose , D-xylose, D-glucose, D-mannose, D-fructose, D-galactose, mal Over vinegar, sucrose, lactose,
It produces acid but not gas from trehalose, D-mannitol and starch. D-sorbitol, inositol and glycerin do not produce any acid or gas. (20) Nutrition requirements None

[0013] The above results can be obtained by
Of the systematic bacteriology. This strain is Gram-positive, bacillus, catalase and oxidase-positive, has motility, and is a bacterium belonging to the genus Bacillus because it forms spores.
There is no similar species in facultative anaerobic, spore shape, swelling position, assimilation of various sugars, salt concentration tolerance, etc. (Biotechnological Laboratory No. 16921).

The new strain differs from other strains belonging to the genus Bacillus in the following points in bacteriological properties. The most closely related species to this new strain is Bacillus pasteurii.
Different in starch hydrolyzability, growth at 55 ° C.
The next closest is Bacillus licheniformis, which differs in spore shape, spore swelling, and growth below pH 6.8. Bacillus coagulrans grow + at 55 ° C, but differ in spore morphology, liquefaction of gelatin, growth below pH 6.8, and salt concentration tolerance. Bacillus circulans differ in spore morphology, VP test, growth below pH 6.8, salt concentration tolerance, and growth at 55 ° C. Bacillus brevis
Indicates the spore shape, growth under anaerobic conditions, generation of acid from sugar, VP
Tests, growth at pH 6.8 or lower, salt concentration tolerance are different.

In practicing the present invention, the present bacterium may be directly inoculated into a dye-containing solution, or cells previously cultured may be added to the dye-containing solution. The cultivation is not particularly limited, but various carbon sources such as various glucoses, carbon sources such as proteins, peptones, yeast extracts, and amino acids, and various inorganic salts such as ammonium sulfate, sodium nitrate, magnesium sulfate, and ferrous sulfate. And the like can be used. The pH and temperature conditions in the culture are as described above. Either aerobic or anaerobic may be used, but the growth rate is higher under aerobic conditions.

The temperature for the decolorization treatment using the present invention is not particularly limited.
C. or less is preferred. Considering utilization at an optimum treatment rate and prevention of contamination by various bacteria, 40 to 55 ° C is more preferable. Also p
Regarding H, the pH is preferably 7.5 or more and 11.0 or less, and more preferably 8.5 or more and 10.5 or less.

The material to be treated is not particularly limited, and examples thereof include an aqueous solution containing a dye that is decolorized by the fungus, factory wastewater, and sewage. Considering that the bacterium has a decolorizing ability particularly under alkaline, medium and high temperature conditions, it has a remarkable effect on the decolorization of the dyeing waste liquid by the reactive dye which is dyed under the same conditions.

Although this strain is facultatively anaerobic, it does not decolorize under aerobic conditions, and its decolorization speed is slightly reduced even under completely anaerobic conditions. Therefore, it is most preferable to perform the reaction in a moderately anaerobic state, and it is not necessary to supply oxygen as in the aerobic condition.
Economically advantageous.

Hereinafter, the present invention will be described specifically with reference to examples.

[0020]

[Example 1] Peptone 0.5%, yeast extract 0.2%, dipotassium phosphate 0.1%, magnesium sulfate heptahydrate 0.02%, sodium carbonate 1
% Of reactive black 5 in a basic medium containing 0.025%
%, Solid medium supplemented with 1.5% agar (pH 10.5)
Was prepared, and a microbial cell suspension of Bacillus sp. DA1 was dripped on the surface thereof and cultured at 45 ° C. One day later, in the dark blue medium, an orange clear zone (visible on the opposite side or see through) was formed around the point where the cell suspension was dropped.

Example 2 1% soluble starch, 0.5% peptone, 0.5% yeast extract, 0.1% dipotassium phosphate, 0.0% magnesium sulfate heptahydrate
Bacillus s in a liquid medium containing 2% and 1% sodium carbonate
p. DA1 was inoculated and cultured aerobically at 45 ° C. overnight. The cells obtained by centrifuging 15 ml of this culture were suspended in a basic medium (described in Example 1) containing 0.025% of various dyes, and allowed to stand at 45 ° C. In each dye-containing medium, the case where no cells were added was used as a control. Eight hours later, the supernatant at the maximum absorption wavelength of each centrifuged supernatant was measured for the centrifuged supernatant.
The decolorization rate was expressed as follows. The results are summarized in Table 1. Disperse dyes (Disperse yellow 3 and Disperse blue 3 in the table)
Acetone was added to a final concentration of 50% to prepare a centrifuged supernatant.

[0022]

[Table 1]

[0023]

The use of the novel microorganisms has made it possible to reduce the degree of coloring of a solution colored with a dye under a medium-high-temperature, alkaline condition. In particular, it has a remarkable effect on wastewater treatment of dyeing using a reactive dye under high temperature and high alkali conditions.

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Claims (10)

[Claims] 1. A novel microorganism belonging to the genus Bacillus and capable of decolorizing a dye under alkaline conditions. 2. A novel microorganism belonging to the genus Bacillus and capable of decolorizing a dye at a temperature of 40 ° C. or higher. 3. A novel microorganism belonging to the genus Bacillus and having a decolorizing ability of a reactive dye under alkaline conditions and at a temperature of 40 ° C. or higher. 4. A new strain belonging to the genus Bacillus which differs from other strains in mycological properties in facultative anaerobic, spore form, swelling position, assimilation of various sugars and salt concentration tolerance. Bacillus sp. DAI. 5. A biotechnological laboratory having the ability to decolorize dyes.
Bacillus sp. Strain DA1 deposited as No. 6921. 6. A method for decolorizing a dye, comprising treating the microorganism according to claim 1 under alkaline conditions. 7. A method for decolorizing a dye, comprising treating the microorganism according to claim 2 at 40 ° C. or higher. 8. A method for decolorizing a dye, comprising treating the microorganism according to claim 3 at 40 ° C. or higher under alkaline conditions. 9. A method for decolorizing a dye, comprising using a new strain of Bacillus sp. DAI. 10. The decolorizing method according to claim 9, wherein the new strain Bacillus sp. DAI is a Bacillus sp. DAI strain deposited as Seiko No. 16921.