JP2000233197A - Dye separating method using dye adsorbable microorganism - Google Patents

Abstract

PROBLEM TO BE SOLVED: To separate and remove a dye from a dye-containing waste soln. by using a microorganism to reutilize the recovered dye. SOLUTION: A method for separating a dye from a dye-containing aq. liquid has a process for bringing a microorganism having capacity adsorbing or taking in the dye into contact with the dye-containing aq. liquid and a process for separating the microorganism adsorbing or taking in the dye from the aq. liquid and, as the microorganism usable in this method, Aspergillus fumigatus, Penicillium janthinellum, Agrobacterium radiobacter, Pseudomonas putida and Pseudomonas sp. strain CDC group 1 are designated.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention provides a novel method for treating wastewater containing dyes using microorganisms.

[0002]

2. Description of the Related Art Conventionally, as a method of removing a dye from a waste liquid using a microorganism, Japanese Patent Application Laid-Open No. 48-95063 discloses Sa.
The method of removing heavy metal compounds, dyes and synthetic detergents using ccharomyces yeast is described in JP-A-6-142665, and the use of Pseudomonas bacteria removes persistent chemicals such as dyes, pigments and surfactants. Only the method is disclosed. Here, Pseudomonas cepacia used in the examples of JP-A-6-142665 is a bacterium that has been identified in recent studies as belonging to the genus Burkholderia instead of the genus Pseudomonas (Yabuuchi et al., Microbio
l . Immunol ., 36 (12) 1251-1275, 1992 and Gillis et al., I.
nt. J. Syst. Bacteriol. 45 (2), 274-278, 1995). Accordingly, JP-A-6-142665, paragraph number 0010, Ps
The statement that the genus eudomonas is useful for removing dyes is actually based on the results of an example performed using a bacterium belonging to the genus Burkolderia , and is a statement without any support.

For other microorganisms, there is no prior art which can be used to treat the dye or even recover the dye in a usable form.

[0004]

SUMMARY OF THE INVENTION An object of the present invention is to provide a method for separating and removing a dye from a waste liquid containing the dye by using a microorganism, and for recycling the recovered dye.

[0005]

SUMMARY OF THE INVENTION The present invention comprises the steps of contacting a microorganism capable of adsorbing or incorporating a dye with an aqueous liquid containing the dye and separating the microorganism adsorbing or incorporating the dye from the aqueous liquid. And a method for separating a dye from an aqueous liquid containing the dye.

[0006]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS A microorganism capable of adsorbing or incorporating a dye which can be used in the method of the present invention can survive in the presence of the dye to be treated, and further adsorbs or absorbs the dye to be treated. It can be easily obtained by selecting a microorganism having the ability to take up.

[0007] One example of a screening method for selecting a microorganism that can be used in the present invention is an appropriate concentration, for example, 0.1%.
The microbial isolate is diluted and cultured in a medium containing 01-0.1% of the dye to be treated. The obtained culture is cultured on a plate medium containing the same dye,
Select strains that form distinct colonies. The selected strain is transferred to a dye-containing plate medium, and a strain in which the color of the dye around the colony has disappeared is selected. The selected strain is inoculated into a dye-containing liquid medium, and a strain that loses the color of the dye with culturing is selected as a microorganism capable of adsorbing or incorporating the dye that can be used in the present invention.

[0008] Specific examples of the microorganism that can be used in the present invention include microorganisms belonging to the genus Aspergillus , such as Aspergillus f.
umigatus , especially Aspergillus fumigatus FERM P-17121,
Microorganisms belonging to the genus Penicillium , such as Penicillium janth
inellum, especially Penicillium janthinellum FERM P-1712
2.Bacteria belonging to the genus Agrobacterium , such as Agrobacteriumr
adiobacter, especially Agrobacterium radiobacter FERM P-17
122, and bacteria belonging to the genus Pseudomonas , such as Pseudomo
nas putida, especially Pseudomonas putida FERM P-17123 and Pseudomonas sp .strain CDC group 1, especially Pseudomo
nas sp . strainCDC group 1 FERM P-17124 is exemplified. These microorganisms may be used alone or in combination of one or more.

The dye that can be treated by the method of the present invention is not particularly limited, and examples thereof include a reactive dye, a vat dye, and a disperse dye. In particular, according to the present invention, those in which the reactive group is a heterocyclic system, such as a dye having a monochlorotriazine moiety, or those in which the reactive group is an acid derivative, such as a reactive dye having a sulfatoethyl sulfone moiety, are preferred. Can be processed.

The aqueous liquid that can be treated by the method of the present invention may contain, in addition to dyes, anything that does not inhibit the growth of microorganisms. Also, dilution if necessary,
Adjust pH and supplement nutrients of microorganisms. Examples of the nutrient include a carbon source, a nitrogen source, and an inorganic substance of a microorganism.

In the method of the present invention, the method for bringing the microorganisms into contact with the aqueous liquid is not particularly limited, but the microorganisms grown in a suitable medium are added to the aqueous liquid, and the microorganism-adsorbed carrier is added. Is added to the aqueous liquid. The medium and culture conditions for growing the microorganisms may be appropriately selected as appropriate for each microorganism. Media suitable for various microorganisms and media for culturing filamentous fungi, bacteria and the like are well known to those skilled in the art.

In the method of the present invention, the amount of microorganisms to be added to the aqueous liquid may be appropriately selected, but is preferably 0.01 g to 1000 g, preferably about 1 g per liter of the liquid to be treated.
Of microorganisms. Conditions for contact with microorganisms are as follows:
The microorganism to be used, the composition of the liquid, the properties of the dye to be treated, etc. may be appropriately selected, but typically,
The aqueous liquid to which the microorganism has been added is heated, for example, to a temperature of 10 to 50
2 ° C., preferably 20-37 ° C. with slow stirring.
The treatment may be performed for 10 to 10 days, more preferably 3 to 5 days.

After completion of the treatment, microorganisms that have absorbed or taken up the dye from the liquid to be treated are separated. The microorganisms may be separated by any method usually used for separating microorganisms from a culture solution, such as centrifugation, filtration, membrane separation, and precipitation.

The present invention further provides a method for recovering a dye adsorbed on or taken up by a microorganism. In order to recover the dye, microorganisms having adsorbed or taken up the dye may be separated from the liquid, cells may be destroyed, and the released dye may be separated from the cell residue. As a method for destroying cells, a method well known to those skilled in the art may be used, for example, physical methods such as ultrasonic waves and osmotic shock, and chemical methods such as lysis of cells using detergents and enzymes. An exemplary method is illustrated.

After disrupting the cells, the resulting dye and cell debris are separated. Separation may be performed by any conventionally known method such as centrifugation, filtration, membrane separation, and the like. The recovered dye can be appropriately diluted as it is and used for dyeing, but may be used after purification if necessary.

The dye recovered by the method of the present invention can be appropriately diluted and reused in the subsequent dyeing, like the original dye.

[0017]

Example 1 Screening Microorganisms that can be used in the present invention were selected by screening as follows. The medium used is Table 1:

[Table 1] As shown in FIG.

1 ml of activated sludge and soil samples collected from the wastewater treatment facility were suspended in 9 ml of sterilized saline, and 50 μl of the suspension was placed in a 50 ml test tube in a medium A (10 ml). ) And inoculated at 30 ° C. in the dark under 250
The cells were cultured with shaking at rpm. 100 μl of culture solution every week
Then, the cells were subcultured into 10 ml of a fresh medium and cultured for a total of 3 weeks. The obtained culture solution was inoculated with a platinum loop onto a plate medium of Medium A, and cultured statically at 30 ° C. in the dark.

Twenty colonies forming a clear colony were selected, subcultured on a plate medium of medium B, and further cultured under the same conditions. Media A and B are both reactive blue dyes, C.I.
I. It contains 0.2 g / L of Reactive Blue 19, which is a medium that can be clearly recognized as deep blue by the naked eye. C. I. Reactive Blue 19 is a dye having a sulfatoethyl sulfone moiety as a reactive group. Two
Five days later, the strain around the colony has become pale blue,
Alternatively, decolorized strains were visually selected and 15 strains were selected. The selected strains were inoculated into the liquid medium of Medium B in Table 1, and after culturing for 2 to 5 days, five strains were selected by visual observation where the blue color of the medium had disappeared or became pale.

The obtained strains were designated KB006 and KB, respectively.
007, KB009, KB010 and KB011, were identified as follows based on the results of microbiological observation of each strain.

KB006: A velvet-shaped late-spreading colony is formed on a Tzapek Dox medium agar medium. The colony surface initially appeared white and later turned dark green. The back of the colony was white for both strains. The conidiophores form an enlarged apical sac at the apical end and line up parallel to the conidia. The cell wall became thicker, the conidiophores formed without branching from the specialized podocytes, and released the pigmented conidia from the single-row phialide. Conidia diameter is 2.5-3.
0 μm, colonies on Tzapek Dox agar
After culturing at 25 ° C, the diameter became 7 cm in 10 days. The growth of the obtained colonies was confirmed even at 40 ° C.

From the results of the above microbiological observations, the strain was identified as Asperg
illus fumigatus was identified. This strain was deposited at the Research Institute of Microbial Industry and Technology, Ministry of International Trade and Industry (Deposit date: December 24, 1998, accession number FERM P-17121).

KB007: A colony having a powdery surface layer was formed on a Tzapek Dox agar medium. The colony surface initially appeared white and became light-grey-green over time, but the periphery of the colony remained white. The back of the colony was white.

The conidiophores diverged and exhibited subsequence with metulas supporting phialide. The phialides are arranged on a spiral and the conidia are in series and do not branch. The conidium pattern and the whole conidium showed a broom shape. The conidia had an elliptical shape, had a protruding end, and had a length of 3.0 to 3.5 μm. Colonies on Tzapek Dox agar grew to 6 cm after 10 days of culture.

From the results of the above microbiological observations, the strain was
llium janthinellum was identified. This strain was deposited with the Research Institute of Microbial Industry and Technology of the Ministry of International Trade and Industry (Deposit date: December 24, 1998, accession number FERM P-17122).

KB009, KB010, KB011 Bacteriological observations are as follows:

[Table 2]

[0027]

[Table 3] From the observation results shown in the table, KB009 is Agrobacterium
radiobacter , KB010 is Pseudomonas putida , KB
011 was identified as Pseudomonas sp. Strain CDC group 1. Each strain was deposited at the Research Institute of Microbial Technology, the Ministry of International Trade and Industry. FERM P-17123 (KB0
09), FERM P-17124 (KB010) and FERM P-17
A deposit number of 125 (KB011) was obtained.

[0028]

Example 2 Reactive dye of medium B I. A medium was prepared in which the concentration of Reactive Blue 19 was 0.01%, and the absorption spectrum at 350 to 800 nm was measured using a spectrophotometer. 10 ml of this medium was dispensed into a 50-ml test tube, and one loop of the KB006 strain or KB007 strain was inoculated and cultured with shaking at 30 ° C. for 2 days. After the culture, each culture was filtered with a No. 2 filter paper, and the absorption spectrum of the obtained filtrate at 350 to 800 nm was measured. The measurement was performed with a double beam spectrophotometer 220 (manufactured by Hitachi, Ltd.). FIGS. 1 and 2 show the results of absorption spectrum measurement before and after the treatment with each microorganism. Before the treatment, the absorption observed at around 590 nm almost disappeared after the treatment.

[0029]

Example 3 The ability to treat dyes of each color was examined using the five isolated strains. All dyes were purchased from Sumitomo Chemical Co., Ltd. Dyes used: 1 Yellow: Sumifix Supra Yellow 3RF 2 Black: Sumifix Black B3 Red: Sumifix Supra Scarlet 2GF 4 Blue: Sumifix Supra Blue BRF These are all reactive dyes. There is a monochlorotriazine moiety as a reactive group other than the black dye of 2,
The black dye has a sulfatoethyl sulfone moiety as a reactive group.

Medium used Yeast extract 1 g / L Malt extract 10 g / L Glucose 4 g / L pH 7

Preparation of microorganisms: One loopful of each isolated strain was added to 5 ml of medium, and the mixture was added at 25 ° C. and 200 rpm.
For 1 day with shaking.

Preparation of dye-containing medium: Each dye was added to the medium at a concentration of 0.5%.
It was added so as to be 02%, and this was dispensed into a 500 ml Sakaguchi flask by 200 ml at a time. The dispensed medium was subjected to autoclave sterilization and subjected to the following tests.

Dye removal test: 1 ml of bacterial suspension after culture
Was added to the above medium, and cultured with shaking at 25 ° C. and 200 rpm for 1 week. After the culture is completed, the culture solution is set at 1000 rpm.
And the supernatant was filtered through a 0.2 μm membrane filter. The obtained filtrate was visually observed, and the filtrate was diluted with distilled water to 1/5. With respect to the diluted solution, an absorption spectrum of visible light of 350 to 900 nm was measured. As a control, each dye was added at 0.
The absorption spectrum of a medium that had been treated in the same manner except that the cells were added to a medium containing 02% was measured.

The absorption spectra of the dyes of each color are shown in FIGS. As a result of visual observation, when treated with KB007, the colors of all dyes disappeared, and the treatment liquid became almost colorless and transparent. The treatment liquid with KB006 changed color and turned brown. This is a component of the dye used, ie KB0
It is considered that the components easily absorbed by 06 were preferentially removed. In KB009 and KB010, the color of the original dye remained, but the color was fading. K
In B011, almost the same color as the control remained after one week of treatment. The results of these visual observations are shown in FIGS.
The data almost agreed with the visible light spectrum data described in No. 6.

[0035]

Example 4 Each of dyes 1 to 4 used in Example 3 and
C. I. Reactive Blue 19 (5 blue) was added to each.
1.5 g / L of agar was added to a medium containing 02% (w / v) to prepare a solid medium. Each of the above five strains was inoculated on each solid medium, and cultured at 25 ° C. in the dark for 10 days. The color of the cells grown on the solid medium and the color of the solid medium around the cells were visually observed. Table 4 shows the results.

[0036]

[Table 4]

[0037]

Example 5 KB007 strain was cultured in the same dye-containing medium as in Example 3 under the same conditions as in Example 2 for 10 days.
Filtration was performed with a No. 2 filter paper, and the cells on which the dye was adsorbed or incorporated were collected. Each cell was suspended in 10 ml of distilled water and treated with an ultrasonic cleaner for eyeglasses for 5 minutes. The obtained treatment liquid was filtered through No. 2 filter paper to obtain a filtrate.

The obtained filtrate was diluted to 0.02% with distilled water, and 100 ml of each was diluted with a 10 cm × 10 cm cloth (A).
I2139 bleaching; Kurashiki Spinning Co., Ltd.) at 90 ° C.
Stained at 100 rpm for 4 hours. Each of the obtained fabrics had a vivid color similar to that obtained by dyeing with a normal dye. The hue of each cloth was measured with a spectrolino manufactured by Gretag. Table 5 shows the results.

[0039]

[Table 5] When the respective dyes were used as such and dyed as above, these values were as follows:

[0040]

[Table 6] Each value was almost the same between the recovered dye and the original dye, indicating that the dye recovered by the method of the present invention can be dyed as usual.

[Brief description of the drawings]

FIG. 1 shows a reactive dye with KB006 in Example 2.
C. I. 5 is a chart showing absorption spectra before and after the treatment of an aqueous liquid containing Reactive Blue 19.

FIG. 2. In Example 2, KB007 is a reactive dye.
C. I. 5 is a chart showing absorption spectra before and after the treatment of an aqueous liquid containing Reactive Blue 19.

FIG. 3 is a chart showing an absorption spectrum in the case where an aqueous liquid containing S. fixus Supra Yellow 3RF is treated using five types of microorganisms in Example 3.

FIG. 4 is a chart showing an absorption spectrum in the case where an aqueous liquid containing Sumifix Black B is treated using five types of microorganisms in Example 3.

FIG. 5 is a chart showing an absorption spectrum in the case of treating an aqueous liquid containing Sumifix Supra Scarlet 2GF using five types of microorganisms in Example 3.

FIG. 6 is a chart showing an absorption spectrum in the case where an aqueous liquid containing Sumifix Supra Blue BRF is treated using five types of microorganisms in Example 3.

 ────────────────────────────────────────────────── ─── Continued on the front page (72) Inventor Ryohei Yamamoto 14-5 Shimogita-cho, Neyagawa-shi, Osaka Kurashiki Textile Co., Ltd. F-term (reference) 4B065 AA11X AA44X AA60X AA67X AC12 AC20 BA22 CA55 CA56 4D040 DD01 DD11

Claims (14)

[Claims] 1. A method comprising: contacting a microorganism of the genus Aspergillus having the ability to adsorb or take up a dye with an aqueous liquid containing the dye; and separating the microorganism having adsorbed or taken up the dye from the aqueous liquid. A method for separating a dye from an aqueous liquid containing the dye. 2. The method according to claim 1, wherein the microorganism is Aspergillus fumigatus . 3. The method according to claim 1, wherein the microorganism is Aspergillus fumigatus FERM P.
The method of claim 2, wherein the method is -17121. 4. A method comprising: contacting a microorganism of the genus Penicillium having the ability to adsorb or take up a dye with an aqueous liquid containing the dye; and separating the microorganism having adsorbed or taken up the dye from the aqueous liquid. A method for separating a dye from an aqueous liquid containing the dye. 5. The method according to claim 4, wherein the microorganism is Penicillium janthinellum . 6. The method according to claim 6, wherein the microorganism is Penicillium janthinellum FER.
6. The method of claim 5, which is MP-17122. 7. A method comprising: contacting an Agrobacterium microorganism having the ability to adsorb or take up a dye with an aqueous liquid containing the dye; and separating the microorganism having adsorbed or taken up the dye from the aqueous liquid. A method for separating a dye from an aqueous liquid containing the dye. 8. The method according to claim 8, wherein the microorganism is Agrobacterium radiobacter.
The method of claim 7, wherein 9. The method according to claim 9, wherein the microorganism is Agrobacterium radiobacter FE.
9. The method of claim 8, which is RM P-17123. 10. A method comprising: contacting a microorganism of the genus Pseudomonas having the ability to adsorb or take up a dye with an aqueous liquid containing the dye; and separating the microorganism having adsorbed or taken up the dye from the aqueous liquid. A method for separating a dye from an aqueous liquid containing the dye. 11. The method according to claim 11, wherein the microorganism is Pseudomonas putida or
Claim 1 which is Pseudomonas sp. Strain CDC group 1.
0. The method of claim 0. The microorganism may be Pseudomonas putida FERM P-
17124 and Pseudomonas sp .strain CDC group 1 FERM
12. The method of claim 11, wherein the method is selected from the group consisting of P-17125. 13. A strain capable of growing on a medium containing a reactive dye or a vat dye and having the ability to adsorb or take up the reactive dye or the vat dye is brought into contact with an aqueous waste solution containing the dye, and A method for separating a dye from an aqueous liquid containing a dye, comprising a step of separating the adsorbed or incorporated microorganisms from the aqueous liquid. 14. The method according to claim 1, further comprising the step of destroying the separated microorganisms and separating the adsorbed or incorporated dye from the microorganisms.