JP4425033B2 - Purification system and purification method for pollutants containing dioxins, and preparation for decomposing dioxins - Google Patents

Description

  The present invention uses a pollutant purification system and a purification method for decomposing dioxins in a pollutant containing dioxins in an aqueous medium by a microbial cell fragment or a fraction thereof, and for purifying the pollutant. The present invention relates to a preparation for decomposing dioxins.

  Currently, the operation and management of facilities that generate dioxins such as incinerators are regulated by law, and the generation of dioxins is prevented. However, existing facilities that do not meet the regulation values of the law require work such as dismantling and improvement. In such operations, contaminants containing dioxins are generated in various forms. As a preferred technique for decomposing dioxins in such contaminants to purify the contaminants, a technique for decomposing dioxins using microorganisms is known.

  Examples of techniques for decomposing dioxins using microorganisms include, for example, a primary process in which dioxins in a dioxin-containing liquid are adsorbed or sorbed to a solid, captured and concentrated, and dioxins are concentrated. A secondary step of separating the treatment liquid containing the solid into solid and liquid to obtain separated water from which dioxins are removed and a concentrated slurry; and a tertiary step of biologically treating the concentrated slurry to decompose dioxins in the concentrated slurry. In the biological treatment in the third step, there is known a method for treating a dioxin-containing liquid using a wood decay fungus or / and a production enzyme of the wood decay fungus (for example, see Patent Document 1). .

  In addition, as a technique for decomposing dioxins using microorganisms, for example, an incinerator facility contaminated with dioxins is shut off from the surrounding environment, and the inside surface of the incinerator facility has a dioxin-decomposing microorganism. Dioxin contamination characterized by adhering a composition containing a film-forming immobilizing agent and dismantling the incineration facility, and using wood-rotting fungi such as white-rotting fungi of basidiomycetes as preferred microorganisms A structure demolition method is known (for example, see Patent Document 2).

  In addition, as a microorganism that degrades dioxins, Bacillus midousuji (for example, see Non-patent Document 1) is known, and a contaminant purification technique using Bacillus midusuji is known. (For example, see Patent Document 3).

  However, in the conventional technique, the decomposition activity may be lost for dioxins having 3 or more chlorinations depending on the bacteria used and the production enzymes. This is thought to be due to the steric hindrance caused by chlorine bound to the skeleton structure of dioxins, and the enzymes produced by the bacteria cannot act on the degradation of dioxins such as dechlorination and are deactivated.

At present, the law also stipulates the handling of contaminants containing dioxins. For this reason, it is desired to decompose dioxins at the site where the contaminant is generated without transporting the contaminant to a facility such as a treatment plant. However, when dioxins are decomposed using fungi at the site where the contaminants are generated, an environment where the bacteria can survive in the site where the contaminants are generated or during the purification process of the contaminants (adjustment of temperature, pH, salt concentration and medium) Etc.), and there remains room for study in terms of simplifying the purification system and improving efficiency.
JP 2002-28695 A JP 2001-90353 A Sadayo Hoshina, 3 others, “Dioxin degradation experiments and gene analysis by thermophilic bacteria”, Proceedings of the 10th Annual Conference of the Waste Society of Japan, Waste Society of Japan, October 10, 1999, p. 883-885 JP 2002-301466 A

  The present invention is capable of decomposing dioxins having three or more chlorinations in pollutants, and easily purifies pollutants at a site where dioxins are generated such as cleaning operations and demolition operations of incineration facilities. This is the issue.

  The present inventors have developed Bacillus midousuji (hereinafter referred to as Bacillus midousuji), which has been cultured in the presence of a chlorinated aromatic compound having a substituent having an oxygen atom bonded to an aromatic ring and a chloro group bonded to the aromatic ring. It was found that a crushed cell body containing a cell membrane of “midosuji” (also called “midosuji”) also decomposes dioxins having 3 or more chlorinations, and has led to the present invention.

  That is, the present invention is a system for purifying contaminants by decomposing dioxins in a contaminant containing dioxins, and a substituent having an oxygen atom bonded to an aromatic ring and a chloro group bonded to the aromatic ring. Bacteria crushed or a fraction thereof containing a cell membrane of Bacillus midusuji cultured in the presence of a chlorinated aromatic compound having the following structure (hereinafter referred to as "bacteria crushed or fraction thereof" A pollutant purification system having a reaction tank containing at least a contaminant and an aqueous medium.

  As is apparent from the examination results described later, the disrupted cell body containing a cell membrane of Midosuji cleaves the characteristic ether bond of dioxins. Therefore, according to the said structure, it is possible for the said microbial cell crushed material etc. and the said contaminant to decompose | disassemble dioxins irrespective of the chlorination number in a reaction tank. In addition, according to the above configuration, since the crushed cells are used, it is necessary to maintain facilities for maintaining the survival of microorganisms and the activity of microorganisms and maintaining a strict environment as compared with purification technologies using microorganisms. And not. Therefore, management of purification of contaminants is easy, and contaminants can be easily purified.

  Moreover, it is preferable that the purification system of this invention has a filtration means for isolate | separating an aqueous medium and a solid substance from the accommodation in a reaction tank, and taking out an aqueous medium. According to such a configuration, only the aqueous medium can be taken out from the contents in the reaction tank, and the concentration of contaminants in the contents in the reaction tank can be easily controlled. It is preferable to control the contact between the contaminants and the contaminants and increase the purification efficiency of the contaminants.

  In addition, the purification system of the present invention includes an isolating means for isolating the source of contaminants, and a fluid generating means for generating a fluid containing the contaminants by including the contaminants of the contaminant source in at least water. It is preferable to have fluid conveying means for conveying a fluid containing contaminants toward the reaction tank. According to such a configuration, it is possible to prevent the contaminants from being scattered to the surroundings from the source of the contaminants, and to prevent the contaminants from being scattered during the transportation of the contaminants. It is preferable for purifying contaminants containing dioxins at the site such as soil improvement on the site of the facility.

In the purification system of the present invention, it is preferable that the fluid generating unit is a contaminant cleaning unit that injects at least water to a contaminant generation source to wash away the contaminant. According to such a configuration, when the incinerator is dismantled or the incineration facility is improved, the cleaning wastewater from the contaminants attached to these facilities is used as it is for purification of the contaminants. It is preferable for purifying contaminants on site according to the configuration.

  The present invention also relates to a method of decomposing dioxins in a contaminant containing dioxins to purify the contaminant, and a substituent having an oxygen atom bonded to the aromatic ring and a chloro group bonded to the aromatic ring. The present invention provides a method for purifying contaminants, which comprises mixing a disrupted cell body containing a cell membrane of Bacillus midusuji cultured in the presence of a chlorinated aromatic compound containing the contaminant and an aqueous medium.

  According to the said method, since the said microbial cell crushed material etc. and the said contaminant contact, it is possible to decompose | disassemble dioxins irrespective of chlorination number. Moreover, according to the said method, since the said microbial cell crushed material etc. are used, the installation for maintaining the survival of microorganisms and the activity of microorganisms is not required compared with the purification technique using microorganisms. Therefore, it is easy to manage the conditions for the purification of contaminants, and the contaminants can be easily purified.

  Further, the purification method of the present invention separates the solid and the aqueous medium from the mixture and takes out the aqueous medium from which the solid has been removed, thereby easily controlling the concentration of contaminants in the mixture, This is preferable for improving the purification efficiency of contaminants by controlling the contact between the crushed material and the contaminants.

  In the purification method of the present invention, the source of contaminants is isolated, the contaminants generated from the isolated contaminant sources are included in water, and the disrupted cells are mixed with the water containing the contaminants. Prevents the scattering of pollutants from the source of the pollutants to the surroundings, and the scattering of pollutants during the transportation of the pollutants, and purifies the pollutants at the site such as dismantling of the incinerator and soil improvement at the site of the incineration facility Preferred above.

  Further, the purification method of the present invention is a high-pressure water washing method in which water is sprayed at a high pressure to the pollutant source to wash away the pollutant, or water and abrasive grains are jetted at a high pressure to the pollutant source. Mixing crushed cells with water slurry containing pollutants discharged by the wet sandblasting method, which is washed away, can be used to clean the wastewater from pollutants during incinerator demolition and incineration facility improvement work. It is preferable for purifying contaminants on site with simple equipment.

  The present invention purifies pollutants by decomposing dioxins in pollutants containing dioxins. “Dioxins” is a general term for all of polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and coplanar PCBs (polychlorinated biphenyls). In the present invention, unless otherwise specified, one of these compounds. Part or all.

  In the present invention, a microbial cell comprising a cell membrane of Midosuji fungus cultured in the presence of a chlorinated aromatic compound having a substituent having an oxygen atom bonded to the aromatic ring and a chloro group bonded to the aromatic ring Crushed materials are used. Bacillus midousuji is a gram-positive gonococcus, a thermophilic bacterium that requires a temperature of 62 ° C. or higher for growth and can grow even at 90 ° C. Specific examples include Bacillus midousuji SH2A strain, SH2B-J1 strain, and SH2B-J2 strain. Such strains of Midosuji are deposited under the accession numbers: ATCC 55926 and ATCC 202050 at the National Institute of Advanced Industrial Science and Technology.

  The Midosuji fungus for obtaining the crushed cell etc. is cultured in the presence of a chlorinated aromatic compound having a substituent having an oxygen atom bonded to the aromatic ring and a chloro group bonded to the aromatic ring. . The chlorinated aromatic compound used in the present invention is not particularly limited as long as it is an aromatic compound having a substituent having an oxygen atom bonded to the aromatic ring and a chloro group bonded to the aromatic ring. Examples of such compounds include chlorinated aromatic compounds that are less toxic than dioxins, such as dioxins and chlorinated phenols.

  In the culture of Midosuji, one or more of the chlorinated aromatic compounds can be used. The form of the chlorinated aromatic compound at the time of culture of Midosuji is not particularly limited, and may be a purified product or a composition containing the chlorinated aromatic compound. good. Examples of the composition containing the chlorinated aromatic compound include fly ash and contaminated soil contaminated with the chlorinated aromatic compound.

  More specifically, the culture of Midosuji is mixed with a dioxin content such as dioxins or fly ash, or the chlorinated phenol or the like in a medium such as a liquid medium that serves as a nutrient source for Midosuji. Then, oxygen is supplied to the medium by blowing air or the like, and the temperature of the medium is controlled to a temperature of 62 ° C. or higher at which Midosuji can act.

  The concentration of the chlorinated aromatic compound in the culture of Midosuji is 1 ng / L to 0.1 ng / L with respect to the medium from the viewpoint of obtaining a crushed cell body having sufficient dioxin-degrading activity. Is preferred. Further, as a medium for culturing Midosuji, a medium usually used for culturing Bacillus bacteria can be used except that dioxins or contaminants containing dioxins are mixed. As a means for culturing Midosuji in the presence of the chlorinated aromatic compound, for example, a contaminant purification device described in JP-A No. 2002-301466 can be used.

  In the culture of Midosuji, other substances suitable for culture of Midosuji can be added. Examples of such other substances include activators such as calcium ions that activate the growth reaction of Midosuji, dimethyl sulfoxide (hereinafter referred to as “DMSO”) that acts to inhibit lysis of Midosuji by inhibiting protease activity. In order to enhance the contact between yeast, and further, Midosuji and dioxins, a carrier such as zeolite particles carrying one or both of these may be used.

  The crushed cell body containing the cell membrane of Midosuji can be obtained by a method usually used for obtaining a crushed cell body containing the microbial cell membrane. As such a method, for example, a step of crushing bacterial cells by ultrasonic wave, pressing, addition of cell membrane degrading enzyme, etc., and separating a cell membrane fraction and a cytoplasm fraction from the crushed product as necessary And a method including a process. As long as the microbial cell disruption product contains a cell membrane of Midosuji cultivated in the presence of the chlorinated aromatic compound, it may contain a crushed product of other parts such as cytoplasm. .

  Further, the fraction of the crushed cell body contains at least a protein having a dioxin-degrading activity, extracts a protein having a dioxin-degrading activity, or a protein having no dioxin-degrading activity. It is obtained by separating. Such protein extraction and separation may be performed by, for example, separation by precipitation, for example, ammonium sulfate precipitation, separation by chromatography, for example, ion exchange chromatography, affinity adsorption chromatography, gel filtration chromatography, etc. , And any combination of these methods.

  In the present invention, the crushed microbial cell can be used as it is mixed with contaminants, etc., and it can also be used as an inorganic carrier such as zeolite, activated carbon, or hydroxyapatite, which has an adsorption ability for organic compounds such as proteins and dioxins. Alternatively, it can be used by being supported on a carrier such as an acrylamide polymer, an organic polymer carrier such as alginic acid or carrageenan in accordance with a commonly used method. Disrupted cells can decompose dioxins in pollutants at temperatures where Midosuji can act, and can also degrade dioxins in pollutants at temperatures lower than those where Midosuji can act. Is possible. It is preferable to use the crushed cell body in a higher temperature environment of 60 ° C. or higher in order to prevent degradation due to contaminants and bacteria in the aqueous medium.

  The contaminant to be purified in the present invention is not particularly limited as long as it contains dioxins. Examples of such contaminants include fly ash, soil containing dioxins, gravel, slurry, liquid, and the like.

  The aqueous medium used in the present invention is not particularly limited as long as it is water or a fluid medium containing water as a main component. Examples of such an aqueous medium include water containing a culture medium, crushed cells of other parts of Midosuji, other additives such as the carrier and pH buffer, and the like.

  The pollutant purification system of the present invention includes at least a pollutant containing dioxins, such as a disrupted cell body containing a cell membrane of Midosuji fungus cultured in the presence of the chlorinated aromatic compound, and an aqueous medium. Has a contained reaction vessel.

  The said reaction tank will not be specifically limited if it is a tank which can accommodate the said contaminants, such as the said microbial cell crushed material, and an aqueous medium.

  In addition, the pollutant purification system of the present invention preferably includes a filtering means for separating the aqueous medium and the solid from the contents in the reaction tank and taking out the aqueous medium. A filtration means will not be specifically limited if it is a form which can collect the solid substance contained in an aqueous medium. The filtering means is selected according to the amount and size of the solid contained in the aqueous medium. As such a filtration means, for example, a polyethylene porous membrane or a hollow fiber membrane having a pore distribution of 50 μm to 0.03 μm is used, and a cylindrical shape in which both ends of the cylinder are closed by such a membrane is formed. Things. By submerging such filtering means in the contents in the reaction tank, only the aqueous medium is discharged into the cylinder. The aqueous medium inside the cylinder is discharged from the inside of the cylinder continuously by a pump or at an arbitrary time.

  In the case of using a filtration means, solid matter such as the contaminants and the crushed bacterial cells are collected in the filtration means. Therefore, use appropriate means such as a cylinder or a blower on the surface of the filtration means. It is preferable to supply a gas such as air in order to prevent clogging due to solid matter in the contained material.

  The contaminant purification system of the present invention preferably includes an isolating means for isolating the contaminant source, and a fluid containing the contaminant by containing at least the contaminant of the contaminant source in water. Fluid generating means for generating and fluid conveying means for conveying a fluid containing contaminants toward the reaction tank. In the present invention, the pollutant generation source refers to an aggregate of contaminants, an adhering contaminant, or the like. Examples of the sources of such contaminants include soil containing dioxins, incinerators and peripheral equipment to which fly ash is attached, equipment to which contaminants are attached as a result of processing of contaminants, and the like. .

  The isolation means is not particularly limited as long as it is a means for isolating the source of contaminants and preventing the contaminants from being scattered around. The isolation means can be selected according to the form of the contaminant source. As such isolation means, for example, an airtight sheet that covers the surface of the contaminated soil, an airtight sheet, and an aggregate covering the arched soil that supports the sheet, and a building that covers the structure. Is mentioned. The aggregate only needs to have sufficient strength to form a building, and a known aggregate such as a steel frame can be used as such an aggregate. Further, the separating means may be a slender bag formed of an airtight sheet and sealed with pressurized air, and the slender bag constitutes the roof or wall of the building or the aggregate. Is possible.

The fluid generating means is not particularly limited as long as it is a means for generating a fluid such as a water slurry containing a contaminant by containing the contaminant of the contaminant generation source in at least water. Examples of such fluid generating means include a tank for mixing contaminated soil and an aqueous medium, and a contaminant cleaning means for injecting at least water to a contaminant generation source to wash away the contaminant. Examples of the contaminant cleaning means include means for injecting water at high pressure and means for introducing abrasive grains into the water to be injected and injecting at high pressure.

  The fluid conveying means is not particularly limited as long as it is a means for conveying a fluid containing contaminants toward the reaction tank. An example of such a fluid conveying means is a pump.

  The contaminant purification system of the present invention may have other means other than the means described above. Such other means include, for example, means suitable for increasing the efficiency of purification of contaminants in the reaction tank, and preventing the scattering of contaminants from the source of contaminants isolated by the isolation means. And a means suitable for purifying the aqueous medium discharged from the reaction tank. Any of these other means can be realized using known techniques.

  As a suitable means for increasing the purification efficiency of the contaminants in the reaction tank, for example, a premix tank for preliminarily mixing the contaminants and the crushed bacterial cells, and the contents contained in the reaction tank are mixed. For example, a temperature adjusting means for adjusting the temperature of the contents in the reaction tank having a heating device, and a pollutant discharging means such as a slurry pump for discharging the pollutants in the reaction tank.

  The said heating apparatus will not be specifically limited if it is an apparatus which can heat the accommodation thing in a reaction tank. As such a heating device, for example, an electric heater laid on the bottom and side surfaces of the tank, a jacket composed of a double structure of the side plate of the tank, a jacket made of pipes or tubes, and hot water circulating through the jacket are generated. And a hot water generator to be used.

  Suitable means for preventing the scattering of contaminants from the source of contaminants isolated by the isolation means include, for example, exhaust means for exhausting air inside the isolation means to the outside, Air purifying means for removing contaminants in the air discharged from the air, differential pressure adjusting means for adjusting the pressure inside the separating means to a negative pressure with respect to the pressure outside the separating means, and the like.

  Means suitable for additionally purifying the aqueous medium discharged from the reaction tank include, for example, wastewater treatment means such as an ultrafilter and an activated sludge tank for treating the aqueous medium taken out from the filtration means. Etc.

  The method for purifying pollutants of the present invention is a method for purifying pollutants by decomposing dioxins in pollutants containing dioxins. In the method for purifying pollutants, Bacillus midusuji cultured in the presence of the chlorinated aromatic compound is used. A crushed microbial cell containing a microbial cell membrane, the contaminant and an aqueous medium are mixed.

  The mixing of the microbial cell crushed material and the like, the contaminants and the aqueous medium is not particularly limited in terms of the timing and order of mixing, but in the case of using a filtering means, before supplying air to the surface of the filtering means. Preferably there is. In the present invention, all of the crushed microbial cells, the contaminants, and the aqueous medium may be mixed simultaneously, or these may be mixed in order. The means for performing the mixing is not particularly limited. For example, in the present invention, a mixing means such as a stirring means may be used, or mixing may be performed using air bubbles supplied to the surface of the filtering means.

The mixture is not particularly limited as long as it is a composition in which dioxins in pollutants are mixed so as to be decomposable by crushed cells. Examples of such a mixture include an aqueous slurry containing a large amount of an aqueous medium and a wet powder material containing a small amount of an aqueous medium. In this invention, it is preferable that the said mixture is a water slurry from a viewpoint of preventing scattering of a contaminant and conveying easily.

  In the purification method of the present invention, preferably, the solid and the aqueous medium are separated from the mixture, and the aqueous medium from which the solid is removed is taken out. The method of taking out the aqueous medium from which the solid matter has been removed is not particularly limited. For example, in the present invention, the aqueous medium may be continuously taken out, the aqueous medium may be taken out intermittently, the aqueous medium may be started from the beginning of the supply of the aqueous medium, or the additional supply of the aqueous medium may be performed. Depending on the case, the aqueous medium may be taken out.

  In the purification method of the present invention, preferably, a pollutant source is isolated, a pollutant generated from the isolated pollutant is included in water, and a microbial cell lysate or the like is mixed with the water containing the pollutant.

  When the contaminant is included in the water, there is no need to use any special means, for example, the wastewater from the source of the contaminant or the wastewater from the work site for dismantling the contaminant can be used. . As a method for obtaining water containing contaminants, for example, a high-pressure water washing method in which water is sprayed to the source of contaminants at a high pressure to wash away the contaminants, or water and abrasive grains are sprayed to the source of contaminants at a high pressure. A wet sandblasting method for washing away contaminants can be used.

In the purification method of the present invention, the end point can be the time when the amount of dioxins in the pollutant falls below the regulation value. The amount of dioxins in pollutants can be measured by the GC / MS method defined in JIS K0311 and JIS K0312. However, the enzyme immunoassay described later can be used for dioxins. This is preferable for rapid quantification. For such an enzyme immunoassay, a commercially available analysis kit can be used. Examples of such an analysis kit include Ecoassay Dioxin (ELISA kit, manufactured by Otsuka Pharmaceutical Co., Ltd.). However, in the enzyme immunoassay, since the composition of the aqueous medium affects the detection result, a calibration curve corresponding to the composition of the aqueous medium is prepared in advance, and this is used to quantify dioxins in pollutants. It is preferable to do.
Hereinafter, the enzyme immunoassay will be described.

<Quantitative method of dioxins suitably used in the present invention>
In this quantification method, a biotinylated dioxin solution with a known concentration labeled with biotin and a sample with an unknown concentration are allowed to act on a light-transmitting stationary phase on which a polyclonal antibody that recognizes dioxins is immobilized. Color the labeled dioxin and measure the concentration of dioxins in the sample using a competitive reaction. In this quantification method, 2,3,7-trichlorodibenzo-p-dioxin (2,3,7-TCDD) labeled with biotin is preferably used as the biotin-labeled dioxin.

  Specifically, each well of the stationary phase plate was washed three times with 300 μL of a washing solution using an ecoassay dioxin (ELISA kit, manufactured by Otsuka Pharmaceutical Co., Ltd.), and 50 μL of a biotin-labeled dioxin solution with a known concentration and 50 μL of a quantitative target. Are added to each well, and these solutions are stirred in each well by vibration or the like, and the stationary phase plate is sealed and reacted at 4 ° C. for 20 hours. After the reaction, each well is washed three times with a washing solution, 100 μL of enzyme-labeled streptavidin (peroxidase) is added to each well, and the stationary phase plate is sealed and reacted at room temperature for 2 hours. After the reaction, each well is washed three times with a washing solution, 100 μL of enzyme reaction substrate solution is added to each well, the stationary phase plate is sealed, and allowed to react at room temperature for 20 minutes in the dark. The dispensing time of the enzyme reaction substrate solution is adjusted so that the reaction time of each well is exactly 20 minutes. The absorbance of each well of light having a wavelength of 450 nm is measured. According to this quantification method, the amount of dioxins in the sample can be quantified in the measurement range of 4.1 ng / mL to 1,000 ng / mL. However, the reaction temperature and reaction time vary somewhat depending on the type of dioxins.

When samples containing various known concentrations of dioxins are used as the sample to be quantified,
It is possible to create a calibration curve. In addition, if the sample to be quantified contains dioxins with known concentrations and contains various additives with different concentrations, a calibration curve reflecting the influence of these additives on the measurement results is created. Is possible. Further, when a sample having a known concentration containing different types of dioxins is used as the sample to be quantified, the sensitivity to 2,3,7-TCDD for each type of dioxins is required. According to this quantitative method, 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) has almost the same sensitivity as 2,3,7-TCDD. Therefore, the result of this quantification method can be expressed as 2,3,7,8-TCDD equivalent by multiplying the obtained numerical value by the ratio of sensitivity depending on the type of dioxins.

  The purification method of the present invention can be carried out using the above-described pollutant purification system of the present invention. However, depending on the form of the mixture or the source of the contaminants, other various means may be used. It is also possible to do. As another means applied to the purification method of the present invention, for example, a contaminant purification apparatus described in JP-A No. 2002-301466 can be cited.

  In the purification method of the present invention, when the source of contaminants is soil, the above-described reaction tank is not used, the contaminated soil is isolated by the isolating means, and the microbial cell fragments are contaminated in the isolating means. It is also possible to carry out the process by stirring the contaminated soil by stirring means such as a screw type stirrer.

  As is clear from the above description, the pollutant purification system and the purification method of the present invention can be used for cleaning or dismantling work before improvement of facilities that generate dioxins such as waste incineration facilities, or for such facilities. The present invention can be applied to various forms of contaminants generated from soil improvement of the former site.

  The present invention also relates to a preparation for degrading dioxins, comprising decomposing a dioxin comprising a crushed cell material containing a cell membrane of Bacillus midusuji cultured in the presence of the chlorinated aromatic compound or a fraction thereof. A formulation is provided. The preparation for degrading dioxins of the present invention can be prepared by crushing the above-mentioned Midosuji fungus body, or extracting or separating proteins from the crushed cell body obtained by crushing the fungus body.

  The preparation for degrading dioxins of the present invention can be used as it is or in a form in which the crushed microbial cells are supported on the carrier described above. The preparation for decomposing dioxins of the present invention is in various conditions such as the form of contaminants containing dioxins, the type of dioxins in the contaminants, the temperature during the decomposition reaction, and the form of the preparation for decomposing dioxins. An appropriate amount is used for the decomposition of dioxins.

  The present invention has a substituent having an oxygen atom bonded to an aromatic ring and a chloro group bonded to an aromatic ring in purifying the contaminant by decomposing the dioxins in the contaminant containing the dioxins. Dioxins with 3 or more chlorinations in pollutants are mixed with crushed cells containing cell membranes of Bacillus midusuji cultured in the presence of chlorinated aromatic compounds, pollutants and aqueous media. Can be decomposed, and contaminants can be easily purified at the site where dioxins are generated such as cleaning and dismantling of incineration facilities.

  Further, in the present invention, when the solid and the aqueous medium are separated from the mixture and the aqueous medium from which the solid is removed is taken out, the contact between the crushed cells and the contaminant is controlled, This is even more effective in increasing the purification efficiency.

Further, in the present invention, when the pollutant source is isolated, the pollutant generated from the isolated pollutant source is included in the water, and the microbial cell crushed material is mixed in the water containing the pollutant, the pollutant source In addition, it is more effective in purifying contaminants containing dioxins at sites such as dismantling of incinerators and soil improvement at the site of incineration facilities. .

  Further, in the present invention, a high pressure water washing method in which water is sprayed at a high pressure to the pollutant generation source to wash away the pollutant, or a wet type in which water and abrasive grains are jetted to the pollutant generation source at a high pressure to wash away the pollutant. When crushed bacteria are mixed with water slurry containing contaminants discharged by the sandblasting method, the contaminants adhering to these facilities are cleaned during incinerator demolition and incineration facility improvement work. Wastewater is used as it is for purification of contaminants, and it is even more effective in purifying contaminants on site with a simple configuration.

  The present invention also relates to a preparation for degrading dioxins, comprising decomposing a dioxin comprising a crushed cell material containing a cell membrane of Bacillus midusuji cultured in the presence of the chlorinated aromatic compound or a fraction thereof. Because it is a pharmaceutical preparation, it is possible to decompose trioxin or higher dioxins in pollutants, and to easily purify pollutants at sites where dioxins are generated, such as cleaning and dismantling of incineration facilities. It is possible to supply a novel preparation for dioxin decomposition that can be performed.

As a pollutant purification system according to an embodiment of the present invention, a purification system with a wastewater purification treatment capacity of 5 m ³ / day is shown below.

  As shown in FIG. 1, the purification system in the present embodiment includes a dismantling and high-pressure cleaning work site 100, a vehicle and equipment cleaning work site 101, and a contaminant purification device 1.

  The dismantling and high-pressure washing work site 100 includes an isolation building 102 that covers a facility where the dismantling and high-pressure washing is performed, a drainage pit 103 in which wastewater generated in the isolation building 102 collects, and drainage of the drainage pit 103 from the vehicle and equipment washing work site 101 And an exhaust device 105 that discharges the air inside the isolation building 102 to the outside.

  The vehicle / equipment cleaning work site 101 includes an isolation building 106 that covers a facility where the vehicle and the equipment are cleaned, a drainage pit in which wastewater generated in the isolation building 106 and dismantling sent from the pump 104 and wastewater from the high-pressure cleaning work site 100 gather. 107, a pump 108 for sending the waste water from the drain pit 107 to the pollutant purification device 1, and an exhaust device 109 for releasing the air inside the isolation building 106 to the outside.

  The isolation buildings 102 and 106 are semi-columnar buildings having a structure in which an aggregate such as a semicircular steel frame is covered with an airtight sheet made of resin or cloth. As the isolation building, it is possible to use a building in which a roof, an aggregate, a wall, or the like is configured by introducing compressed air into an air-tight bag made of resin or cloth and inflated.

  For example, as shown in FIG. 2, the exhaust devices 105 and 109 are provided with a medium performance filter 114, an automatic valve 115, and a fan 117, and are configured by an air discharge passage 118 for discharging the air inside the isolated building to the outside. ing. The automatic valve 115 is connected to, for example, a differential pressure gauge (not shown) that detects a differential pressure between the atmospheric pressure and the atmospheric pressure in the isolated building, and the atmospheric pressure in the isolated building is negative with respect to the atmospheric pressure according to the detection result of the differential pressure gauge. It is a valve that keeps pressure. The amount of exhaust from the isolated building is controlled by, for example, the output of the fan and the opening of the automatic valve.

The contaminant purification apparatus 1 includes a mesh screen 2 for removing solid particles having a relatively large particle size such as abrasive grains and debris from the waste water sent from the pump 108, and a solid material having a large particle size by the mesh screen 2. The raw water tank 3 for storing the waste water from which the water has been removed, the reaction tank 5 for storing the waste water of the raw water tank 3 having a jacket through which the heat medium circulates and being sent via the pump 4, A blower 6 for supplying air, a submerged membrane 7 for taking out the waste water from which the solid matter having a small particle diameter has been removed from the waste water contained in the reaction tank 5, a pump 8, a flow meter 9 and a drain port 10. Air is supplied to the activated sludge tank 12, the activated sludge tank 12 containing the solid matter filtered by the ultra filter 11, and the activated sludge tank 12. Blower 13 and reaction tank 5 jacket A heating device 14 that is connected to the heater and heats the heat medium in the jacket, a pump 15 that sucks solids accumulated at the bottom of the reaction tank 5 and sends it to the activated sludge tank 12, and a reaction tank 5 that has a predetermined water level or higher. And an overflow line 16 for returning the wastewater to the raw water tank 3.

The raw water tank 3 is a tank having a diameter of 2.2 m, a height of 2 m, and a volume of 5 m ³ . The pumps 4 and 8 are pumps having a liquid feeding capacity of 5 m ³ / h. The reaction tank 5 is a tank having a diameter of 3 m, a height of 2 m, and a volume of 10 m ³ . The blower 6 is a blower having an air blowing capacity of 25 m ³ / h. As the heat medium circulating through the jacket of the reaction tank 5, for example, hot water is used as a heat medium capable of arbitrarily adjusting the temperature of the wastewater accommodated in the reaction tank 5.

  The blower 6 supplies air to the bottom of the reaction tank 5 immediately below the submerged membrane 7. The submerged membrane 7 is provided at a position where the reaction tank 5 is submerged when the waste water is accommodated. The submerged membrane 7 is a kind of microfilter, and is a hollow porous membrane having a peripheral surface and end surfaces formed of a polyethylene porous membrane having a pore distribution of 50 to 0.03 μm or a hydrophilic hollow fiber membrane. It is a cylinder.

The ultrafilter 11 is constituted by a polyacrylonitrile copolymer filter having a pore distribution of 0.3 μm to 0.001 μm, for example. The activated sludge tank 12 contains aerobic microorganisms that decompose organic substances such as proteins. The activated sludge tank 12 is a tank having a diameter of 2.2 m, a height of 2 m, and a volume of 5 m ³ . The blower 13 is a blower having a blowing capacity of 1 m ³ / h.

  Next, a method for purifying contaminants containing dioxins generated at the site of dismantling and cleaning work using the above-described purification system will be described. First, preparation of a microbial cell crushed material containing a cell membrane of Midosuji used for purification of contaminants will be described. The microbial cell disruption product is obtained by culturing a large amount of Midosuji in a factory or the like, and crushing and separating the obtained Midosuji fungus.

  For example, an apparatus shown in FIG. 4 is used to prepare the crushed cell body. This apparatus includes a culture tank 20 that stores liquid medium, fly ash, strains, and the like, a draft tube 21 that is submerged in the contents in the culture tank 20, a blower 22 that supplies air to the draft tube 21, and a culture tank 20 The heating medium 23 for heating the heating medium circulating in the jacket provided in the chamber and controlling the temperature of the contents in the culture tank 20 to a temperature at which Midosuji can act, and the exhaust from the culture tank 20 are circulated. A cooler 24 cooled by the refrigerant, a cooling device 25 that cools the refrigerant circulating in the cooler 24, a gas-liquid separator 26 that separates gas and mist from the exhaust gas cooled by the cooler 24, and gas-liquid separation A removal device 27 for removing dioxins in the exhaust gas separated by the vessel 26 from the exhaust gas, an ultracentrifuge 28 for separating a microbial cell lysate including a cell membrane from a microbial cell lysate of Midosuji, Minutes with liquid separator 26 It is constituted by a two-way valve 29 for opening and closing the line to return the liquid in the culture tank 20.

The culture tank 20 is a tank having a diameter of 0.5 m, a height of 1 m, and a volume of 0.1 m ³ . The draft tube 21 is a tube that mixes the contents in the culture tank 20 and air and forms convection of bubbles in the contents in the culture tank 20, promotes gas-liquid contact in the culture tank 20, and cultures It is installed for the purpose of maintaining the maximum dissolved oxygen concentration of the contents in the tank 20 and promoting the stirring of the liquid medium. The blower 22 is a blower having a blowing capacity of 6 m ³ / h. The heating device 23 is a heating device having a heating capacity of 12 kW. The cooler 24 is configured by a plate heat exchanger. The gas-liquid separator 26 is a cyclone type gas-liquid separator. The removal device 27 is an adsorber filled with activated carbon.

First, 0.1 m ³ of water is put into the culture tank 20. Next, air is supplied from the blower 22 to the culture tank 20 through the draft tube 21. The water in the culture tank 20 is heated by the heating device 23 to 65 ° C., which is the optimum temperature for Midosuji. The temperature of the water in the culture tank 20 is 65 ° C. as a control target value, and the flow rate of the heat medium circulating through the jacket is determined by the PID operation or the two-position operation based on the deviation between the measured value of the temperature of the stored item and the control target value. Controlled by.

  When the temperature of the water in the culture tank 20 is maintained at 65 ° C., water vapor and mist generated by the disappearance of bubbles are discharged from the culture tank 20 along with the exhaust. The cooling device 25 cools the refrigerant circulating in the cooler 24 to about 15 ° C., and the discharged water vapor is cooled by the cooler 24 and returned to the culture tank 20 as water. The mist is separated from the exhaust gas by the gas-liquid separator 26 and returned to the culture tank 20 as water. The water accumulated in the gas-liquid separator 26 is returned to the culture tank 20 by opening the two-way valve 29 in conjunction with the stop of the blower 22. When the blower 22 is stopped, that is, the microbial activity in the culture tank 20 is stopped and the two-way valve 29 is opened, the water and liquid medium when the water is returned from the gas-liquid separator 26 to the culture tank 20. Thus, the backflow of the contents in the culture tank 20 is prevented.

  The exhaust gas from which moisture has been removed to some extent by the gas-liquid separator 26 is sent to the exclusion device 27. When the exhaust device contains a small amount of organic substances such as dioxins, the excluding device 27 adsorbs these organic substances and releases the exhaust gas from which these organic substances have been removed to the atmosphere.

When the water in the culture tank 20 is maintained at 65 ° C., the supply of air by the blower 22 is temporarily stopped, and a soybean medium, soy protein, molasses or the like is added to the culture tank 20 to adjust the liquid medium. Next, fly ash as a supply source of dioxins, fresh culture strain of Midosuji, DMSO, Ca ²⁺ , yeast, zeolite, pH adjuster and the like are put into the culture tank 20 and the operation of the blower 22 is resumed. In this example, the Bacillus midusuji SH2B-J2 strain is used as a fresh cultured strain of Midosuji.

  In addition, since the effect of the growth of Midosuji on the concentration of the medium is negligible at a concentration of 3% by mass or more in the culture tank 20 of the medium that is a nutrient source for Midosuji, the medium in the culture medium 20 of the liquid medium The concentration of is 3% by mass or more.

  Further, if the concentration of calcium ions in the culture tank 20 that promotes the energy metabolism of the Midosuji fungus and activates the growth reaction is 50 mmol / L or more, the effect of the growth activity of the Midosuji fungus on the calcium ion concentration is negligible. Therefore, the concentration of calcium ions in the culture tank 20 is set to 50 mmol / L or more.

  In addition, since the concentration of DMSO in the culture tank 20 acting to inhibit the lysis of Midosuji by inhibiting protease activity is 30 mL / L or more, growth inhibition of Midosuji is observed. The concentration is 30 mL / L (3%).

  Hereinafter, the conditions for culturing the SH2B-J2 strain in the presence of dioxins (fly ash) in the culture tank 20 by similarly examining the concentration and various conditions of the various additives in the culture tank 20 and the effects thereof. Is as follows.

<Culture conditions>
Optimal temperature 65 ℃
pH control range 6.0-7.5
Initial dose of Midosuji 1 × 10 ⁷ / mL
Liquid medium concentration 3% by mass or more Ca ²⁺ addition concentration 50 mmol / L or more DMSO addition concentration 30 mL / L (3%)
Zeolite addition concentration 50g / L or more Culture time 3-4 hours Fly ash addition concentration 1g / L

  The aforementioned culture apparatus is operated for 3 to 4 hours to culture Midosuji. The dissolved oxygen concentration in the container in the culture tank 20 and the sugar concentration in the container are measured from the beginning of the culture, and the progress of the culture of Midosuji is determined from these measurement results. As for the dissolved oxygen concentration, when the culture of Midosuji is proceeding normally, a phenomenon is observed in which the dissolved oxygen concentration extremely decreases in the later stage of the logarithmic growth phase. In addition, when the culture of Midosuji is progressing normally, a phenomenon is observed in which the sugar concentration decreases logarithmically with the growth of Midosuji.

  When the end point of the culture of Midosuji is determined from the phenomenon described above, the cells of Midosuji are destroyed. In order to destroy the cells of Midosuji, the cells are destroyed by putting a cell membrane degrading enzyme into the culture tank 20 or by a cell crusher that crushes the cells by ultrasonic vibration or the like. The obtained crushed cells are crushed by the ultracentrifuge 28 from the lower layer, such as fly ash and culture medium, cytoplasmic proteins that are crushed cells containing cytoplasm, and microbial cells. It is separated into cell membrane protein which is a product.

  Next, purification of contaminants containing dioxins discharged with the dismantling and washing of the facility using the above-described contaminant purification apparatus 1 will be described using the obtained crushed bacterial cells.

  In the isolation building 102 of the dismantling and cleaning work site 100, for example, an incinerator that is a source of contaminants containing dioxins is cleaned by a high pressure water washing method or a wet sandblasting method, or the incinerator after washing is wire sawed. Dismantling work is carried out by the construction method, and wastewater containing pollutants containing dioxins is discharged. Waste water from the dismantling and cleaning work site 100 is collected in the drain pit 104 and sent to the vehicle and equipment cleaning work site 101.

  In the isolation building 106 of the vehicle / equipment cleaning work site 101, for example, dismantling, a work machine such as a belt conveyor used at the cleaning work site 100 and a work vehicle such as a bulldozer are washed with water and contain dioxins. Wastewater containing pollutants is discharged. The wastewater from the vehicle / equipment cleaning work site 101 is collected in the drainage pit 108 together with the wastewater from the dismantling / cleaning work site 100 and sent to the contaminant purification apparatus 1.

  The facilities to be cleaned and dismantled at the site of dismantling and cleaning work are isolated from the surroundings by the isolation building 102, and the facilities for cleaning at the vehicle and equipment cleaning work site are isolated from the surroundings by the isolation building 106. ing. Further, the air pressure in the isolation buildings 102 and 106 is maintained at a negative pressure with respect to the atmospheric pressure by the exhaust devices 105 and 109. Accordingly, contaminants such as fly ash containing dioxins are prevented from being released from the isolation buildings 102 and 106 into the atmosphere.

Drainage sent from the vehicle / equipment cleaning work site 101 is supplied to the mesh screen 2. The mesh screen 2 removes solids having a relatively large particle size such as abrasive grains used in the wet sandblasting method, debris discharged during the dismantling operation, and the like from the supplied waste water. The waste water from which such solid matter has been removed is sent to the raw water tank 3. The water level in the raw water tank 3 is measured by an electrode type level meter, and is maintained between a low level and a high level by operating the pump 108. In addition, when the sent waste water is a waste water having physical properties that cause protein denaturation, the physical properties of the waste water may be adjusted in the raw water tank 3 by adding appropriate additives.

  The wastewater sent to the raw water tank 3 is sent to the reaction tank 5 by the pump 4. The water level in the reaction tank 5 is measured by an electrode type level meter, and the waste water supplied excessively to the reaction tank 5 is returned to the raw water tank 3 through the overflow line 16.

  The waste water accommodated in the reaction tank 5 is heated to an appropriate temperature by a jacket to which hot water heated by the heating device 14 is supplied. The reaction vessel 5 is supplied with crushed cell bodies containing a cell membrane of Midosuji cultivated in the presence of the chlorinated aromatic compounds such as dioxins and chlorinated phenols. Air is supplied from the blower 6 to the reaction tank 5. In this manner, in the reaction tank 5, the dioxins are decomposed by the crushed cell bodies.

  In addition, other cell disruptions such as cell disruptions containing cytoplasm obtained during the separation of cell disruptions containing cell membranes, and contaminants such as culture media and fly ash, Other crushed cells obtained by separating the crushed cell material including the microbial cell membrane by supplying the crushed cell material to the reaction tank 5 or supplying other crushed cell materials and impurities to the raw water tank 3 Etc. may be used for the decomposition treatment of dioxins.

  The mixture of the waste water in the reaction tank 5 and the crushed cells is stirred by the air supplied from the blower 6. The air supplied from the blower 6 rises along the surface of the submerged film 7 as fine bubbles. As the decomposition of dioxins by the crushed cells progresses, the mixture becomes brown and increases its color.

  In the reaction tank 5, after carrying out the decomposition reaction of the dioxins with the crushed cell bodies for 18 hours or more, the amount of the dioxins in the mixture in the reaction tank 5 is set to an amount equivalent to 2,3,7,8-TCDD. Measured by immunoassay, confirms that the amount of dioxins sufficiently satisfies the wastewater standard value, and operates the pump 8 to remove inorganic and organic solids of 0.4 μm or more from the mixture by the submerged membrane 7. Filter and discharge to the sewage from the drain 10 as purified water.

  In the filtration of the mixture by the submerged membrane 7, the solid matter in the mixture is filtered into the submerged membrane 7. On the other hand, the blower 6 supplies air from directly under the submerged membrane 7. Since the air bubbles to be moved move upward along the surface of the submerged film 7, clogging of the submerged film 7 is prevented.

  In the case where maintenance of the submerged membrane 7 is performed, or when further clean drainage is required, the drainage sent to the drainage port 10 is sent to the ultrafilter 11. The wastewater sent to the ultrafilter 11 permeates through the membrane while flowing along the surface of the filtration membrane of the ultrafilter 11, and wastewater from which solid matter having a smaller particle size is removed is obtained. The obtained waste water is discharged into sewage as purified water.

  Solid matter (sludge) deposited on the bottom of the reaction tank 5 is supplied to the activated sludge tank 12 by the operation of the pump 15. Further, the solid separated by the ultrafilter 11 is also supplied to the activated sludge tank 12. Air is supplied from the blower 13 into the activated sludge tank 12, and an aerobic environment is maintained in the activated sludge tank 12. In the activated sludge tank 12, organic contaminants such as proteins and culture media are decomposed by the activity of microorganisms. The amount of accumulated sludge in the activated sludge tank 12 is monitored, and the accumulated sludge is appropriately carried out from the activated sludge tank 12. The carried out sludge is regenerated as, for example, eco-cement.

According to the present embodiment, a crushed cell body containing a cell membrane of Bacillus midusuji cultured in the presence of the chlorinated aromatic compound, a contaminant containing dioxins, and an aqueous medium are mixed. . This crushed cell body is considered to be difficult for microorganisms having dioxin-degrading activity, such as white-rot fungi, because it breaks the ether bond that connects two benzene rings, which is a characteristic structure of dioxins. It is possible to decompose dioxins with 3 or more chlorinations.

  Moreover, according to this Embodiment, since the said microbial cell crushed material is used for decomposition | disassembly of dioxins, it is hard to be influenced by purification environments, such as temperature and salt concentration, compared with the purification method using microorganisms. For example, the crushed microbial cells can decompose dioxins at a temperature lower than the temperature at which Midosuji can act, and can also decompose dioxins even in a high salt concentration environment. Therefore, it is possible to easily purify contaminants at the site where dioxins are generated such as cleaning work and dismantling work of incineration facilities.

  Moreover, according to this Embodiment, the said microbial cell crushed material which acts on decomposition | disassembly of dioxins is used for decomposition | disassembly of dioxins. In the case of using microorganisms, the survival and growth of microorganisms affect the purification of contaminants, and dioxins may not be quantitatively decomposed with respect to the amount of added microorganisms. Dioxins can be decomposed quantitatively depending on the amount of crushed cells, and contaminants can be easily purified.

  Further, according to the present embodiment, since the aqueous medium from which the solid matter is removed by the submerged film 7 is taken out, the slurry concentration in the reaction tank 5 can be easily controlled, and contaminants can be efficiently removed. Can be purified.

  Further, according to the present embodiment, the source of the contaminant is isolated from the surrounding environment by the isolation building, and the contaminant is generated in the water inside the isolation building, and this water slurry is contaminated. Since it can be used for purification of substances, a water slurry of contaminants can be easily obtained.

  Further, according to the present embodiment, the source of pollutants is isolated from the surrounding environment by the isolation building, and the generated pollutants are transported in a state of being contained in water. Release to the can be prevented.

  Further, according to the present embodiment, since the atmospheric pressure inside the isolation building is controlled to a negative pressure with respect to the atmospheric pressure, for example, when it is slightly opened to the outside, such as entering and exiting workers, etc. However, the air inside the isolation building does not leak to the outside, and scattering of contaminants to the surrounding environment can be prevented.

  Moreover, according to this Embodiment, since it has the heating apparatus 14, the temperature of the things in the reaction tank 5 can be adjusted suitably at arbitrary timings. Therefore, it is possible to heat the waste water in the reaction tank 5 before putting the crushed cell material into the reaction tank 5 to sterilize the germs in the waste water. Decomposition can be prevented.

  Moreover, according to this Embodiment, since it has the heating apparatus 14, from the viewpoint of improving the dioxin decomposition | disassembly activity of a microbial cell crushed material, the reaction tank 5 at the time of decomposition | disassembly reaction according to the kind etc. of the dioxin of decomposition | disassembly object It is possible to control the temperature of the contained material, and the purified product can be efficiently decomposed.

  In addition, among the microbial cell disruptions of Midosuji fungus cells cultured in the presence of the chlorinated aromatic compound such as dioxins, which part of the microbial cell fragment decomposes dioxins, and dioxins Examinations made by the present inventors regarding which part of the body is decomposed are shown below.

<Examination of degradation sites of dioxins and sites of cells of Midosuji that degrades dioxins>
In order to verify which part of the crushed cell of Midosuji fungus body decomposes dioxins and which part of dioxins is decomposed, the characteristic of dioxins A dioxin-like fluorescent substrate having a unique structure and a crushed product of Midosuji fungus having different culture conditions were reacted under different conditions to detect the presence or absence of a dioxin degradation product under each condition. This method is described in a report by Masaya Nakamura, “Screening for Novel Dioxin Degrading Microorganisms” (Abstracts of Japanese Agricultural Chemistry Conference, P.153, 2001) and Japanese Patent Application Laid-Open No. 2002-348291.

  First, a dioxin-like fluorescent substrate (structural formula A below) having a structure similar to that of dioxins was synthesized. This dioxin-like fluorescent substrate A includes a benzene ring of monochlorobenzene and a fluorescent substance, 6,7-dihydroxy-4-methylchromen-2-one (the following structural formula (C), hereinafter referred to as “esculetin-like”). It is a compound synthesized by connecting the benzene ring of a compound)) with an ether bond, and has a structure having two ether bonds that connect two benzene rings, which is characteristic of dioxins.

  When one of the two ether bonds that bind monochlorobenzene and esculetin C is cleaved, the dioxin-like fluorescent substrate A forms a fluorescent substance B represented by the following structural formula (B), and the two ethers When two of the bonds are cleaved, esculetin-like compound C is formed. When these substances A to C are irradiated with light having a wavelength of 360 nm, as shown in FIG. 5, the dioxin-like fluorescent substrate A hardly emits fluorescence, but the fluorescent substance B and the esculetin-like compound C are excited, It emits strong fluorescence with a wavelength of 450 nm.

  As the Midosuji, a freshly cultured strain obtained by collecting a glycerin-preserving strain on a soybean-Casein Digest Broth agar plate medium with a platinum loop and culturing at 65 ° C. for 3 hours was used. As the glycerin-preserving strain, the first SH2B-J2 strain was isolated from Osaka City in 1995, suspended in glycerin and stored frozen at -85 ° C. The strain was then suspended again in glycerin and cryopreserved at -85 ° C.

  One platinum loop of the fresh cultured strain was collected with a small quantitative platinum loop, and the whole amount was mixed with 15 mL of liquid medium in a 50 mL conical tube. As the liquid medium, 3% by mass of Soybean-Casein Digest Broth and 0.3% by mass of yeast (hereinafter, the liquid medium having this composition is referred to as “liquid medium A”) was used. The conical tube was radially attached to the disk of a disk-type rotary culture apparatus (manufactured by TAITEC), the conical tube was continuously turned over, and a fresh culture solution was mixed and adjusted. The mixing and adjustment of the fresh culture solution by the apparatus was performed under the condition of 2 hours at a rotation speed of 20 rpm in an atmosphere of 65 ° C.

  The obtained 15 mL fresh culture solution and 500 mL of liquid medium A were placed in a 3 L flask, and Midosuji was cultured using a shaker (manufactured by TAITEC). The culture of Midosuji was carried out under the conditions of “8-shaped” rotation in the horizontal direction for 3 hours in a 65 ° C. atmosphere at a rotation speed of 190 rpm. In addition, in the culture of Midosuji, two types of the cultured bacterial solution to which 1 mg / mL fly ash was added and the cultured bacterial solution to which no fly ash was added were used. Fly ash was collected in an incinerator, and this fly ash contains 1 ng-TEQ / g dioxins.

  The obtained culture solution was immediately ice-cooled and centrifuged with a centrifuge (manufactured by BECKMANN). This centrifugation was performed at 4300 G for 20 minutes in an atmosphere of 4 ° C. The obtained bacterial cell pellet was resuspended in an ice-cooled liquid medium (hereinafter referred to as “liquid medium B”), which is a liquid medium obtained by diluting liquid medium A four times, and 50 mL of conical. Housed in a tube.

  The obtained suspension was centrifuged using a centrifuge (manufactured by Sakuma) to obtain bacterial cell sedimentation. This centrifugation was performed in an atmosphere of 4 ° C. under conditions of 10 minutes at a rotation speed of 6,000 rpm.

  The obtained bacterial cell sedimentation was resuspended in 15 mL of liquid medium B, and the resulting suspension was clarified using the ultrasonic crusher (manufactured by Nippon Seiki Seisakusho Co., Ltd.). It was crushed. This ultrasonic crushing was performed four times while cooling with ice at 20 kHz and 150 W for 2 minutes. The crushed suspension was centrifuged using a centrifuge (manufactured by Sakuma) to obtain untreated bacterial cell pellets and supernatant. This centrifugation was performed in a 4 ° C. atmosphere at 6,000 rpm for 10 minutes.

  The obtained supernatant was collected 4 mL each in a test tube for an ultracentrifuge and centrifuged using an ultracentrifuge (manufactured by HITACHI). This centrifugation was performed at 150,000 G for 30 minutes in an atmosphere of 4 ° C. Bacterial cell debris containing cell membranes (hereinafter also referred to as “cell membrane fraction”) is collected from the upper part of the obtained excavation, and the cell debris containing cytoplasm from the lower layer of the obtained excavation ( Hereinafter, it was also referred to as “cytoplasmic fraction”).

  In a 7 mL glass test tube with a screw, 200 μL of the cell membrane fraction or cytoplasmic fraction and 700 μL of liquid medium B are added, and 100 μg of dioxin-like fluorescent substrate A is added to 100 μL of dimethyl sulfoxide (DMSO). DMSO was added so that the final concentration in the reaction solution or the reaction solution was 5% by mass, and allowed to stand at a predetermined temperature for a predetermined time to obtain a reaction solution.

After standing, 12N 50 μL hydrochloric acid was added to the reaction solution, 1 mL of ethyl acetate was added, and the mixture was stirred well and centrifuged. This centrifugation was performed under the condition of 20 minutes at 3,000 rpm. The resulting supernatant ethyl acetate layer was recovered. This operation was repeated three times and evaporated to dryness. The obtained dried product was redissolved with ethyl acetate, 15 μL of this redissolved solution was collected with a glass capillary, spotted on a silica gel plate for thin layer chromatography, and developed with a developing solvent. As the developing solvent, a solvent in which chloroform, ethyl acetate, and formic acid were mixed at a ratio of 10: 8: 1 was used.

  The results of thin layer chromatography are shown in FIG. The conditions of the ethyl acetate solution spotted on each lane of the silica plate shown in FIG. 6 are shown in Table 1 below.

  In FIG. 6, the spot (O) shown at the bottom is the spot position of the redissolved solution, and the large spot (A) shown at the top is the developed dioxin-like fluorescent substrate A. Only in lane 4, spot (X) was observed between spot O and spot A.

  In addition, FIG. 7 shows the results of thin-layer chromatography when the reaction conditions of the cell membrane fraction of Midosuji cultivated with fly ash added and the dioxin-like fluorescent substrate A were changed. Lane 11 in FIG. 7 is a developed redissolved solution when reacted at 18 ° C. for 20 minutes, and lane 12 is developed developed redissolved solution when reacted at 65 ° C. for 18 hours. Is. FIG. 7 shows that a spot X having a higher luminance than that of the lane 11 is observed in the lane 12.

  From the above, it has been clarified that the substance shown in the spot X is a substance having higher polarity than the dioxin-like fluorescent substrate A from the position of the spot X and the conditions of thin layer chromatography. In addition, since the substance shown in the spot X emits strong fluorescence, it is considered that the substance is at least one or both of the fluorescent substance B and the esculetin-like compound C, or a similar compound thereof. Further, since the substance shown in the spot X is detected more in the cell membrane fraction of Midosuji cultivated at 65 ° C., it is generated by the activity of Midosuji cultivated under the condition where fly ash is added. It is considered that the substance acted on.

  Further, when the spot X was measured by the GC / MS method, it was found that the substance shown in the spot X is a substance having a naphthoquinone-like structure and a substance having a sulfate group structure. Therefore, it was revealed that the substance shown in the spot X is a compound similar to the esculetin-like compound C.

  From these results, the cell membrane fraction of Midosuji cultivated in the presence of the chlorinated aromatic compounds such as dioxins is an ether bond connecting two benzene rings, which is characteristic of the structure of dioxins. It became clear to cut.

Further, the cleavage of the ether bond is more remarkable as the activity of Midosuji is more active. Therefore, it is considered that the ether bond is cleaved by an enzyme produced by Midosuji. According to the result of the GC / MS method, since a sulfate group is introduced along with the cleavage of the ether bond, the cleavage of the ether bond is produced by a biological defense reaction in order to render the foreign matter harmless to microorganisms. It is considered that the enzyme is glutathione-s-transferase or arylsulfotransferase.

  In the reaction between the cell membrane fraction and the dioxin-like fluorescent substance A, the reaction solution is generally brown, and the hue increases with the reaction. It is thought to be in progress.

It is a figure showing the whole pollutant purification system composition which is one embodiment of the present invention. It is a figure which shows roughly the structure of the exhaust apparatus used for the purification | cleaning system shown in FIG. It is a figure which shows schematically the structure of the purification apparatus 1 used for the purification system shown in FIG. Cell membrane of Midosuji fungus cultured in the presence of a chlorinated aromatic compound having a substituent having an oxygen atom bonded to an aromatic ring and a chloro group bonded to an aromatic ring, used in the present embodiment It is a figure which shows roughly the structure of the apparatus which prepares the microbial cell crushed material containing this. It is a figure which shows the intensity | strength of each fluorescence with a wavelength of 450 nm when the compound used in examination of the decomposition | disassembly site | part of a dioxin and the kind of microbial cell disruption was excited by the light of a wavelength of 360 nm. In examination of the decomposition | disassembly site | part of a dioxin and the kind of microbial cell crushed material, it is a figure which shows the spot when the reaction liquid obtained on various conditions is spotted and developed on the silica plate. It is a figure which shows a spot when the reaction liquid which changed reaction conditions in the conditions in which the spot X is obtained in the examination of the decomposition | disassembly site | part of a dioxin, and the kind of microbial cell debris is spotted and developed on the silica plate .

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Pollutant purification device 2 Mesh screen 3 Raw water tank 4, 8, 15 Pump 5 Reaction tank 6, 13, 22 Blower 7 Submerged membrane 9 Flowmeter 10 Drain port 11 Ultrafilter 12 Activated sludge tank 14, 23 Heating Equipment 16 Overflow line 20 Culture tank 21 Draft tube 24 Cooler 25 Cooling device 26 Gas-liquid separator 27 Removal device 28 Ultracentrifugal separator 29 Two-way valve 100 Dismantling and cleaning work site 101 Vehicle and equipment cleaning work site 102 and 106 Isolation Building 103, 107 Drain pit 104, 108 Pump 105, 109 Exhaust device 114 Medium performance filter 115 Automatic valve 117 Fan 118 Air discharge passage

Claims (4)

In a method of decomposing dioxins in a contaminant containing dioxins to purify the contaminant,
A cell disruption containing a cell membrane of Bacillus midusuji cultured in the presence of a chlorinated aromatic compound having a substituent having an oxygen atom bonded to the aromatic ring and a chloro group bonded to the aromatic ring, or method for purifying contaminants, wherein Rukoto said and said contaminants and an aqueous medium and the cell membrane fraction containing the fungus membrane, mixed decompose at temperatures lower than 62 ° C..   The method for purifying contaminants according to claim 1, wherein the solid and the aqueous medium are separated from the mixture, and the aqueous medium from which the solid is removed is taken out.   The pollutant source is isolated, the pollutant generated from the isolated pollutant is included in water, and the crushed cell material or a fraction thereof is mixed with water containing the contaminant. Item 3. A method for purifying contaminants according to Item 1 or 2.   Discharge by high pressure water washing method to inject water at high pressure to the pollutant source to wash away the pollutant, or wet sandblast method to inject water and abrasive grains to the pollutant source at a high pressure to wash away the contaminant The method for purifying contaminants according to claim 3, wherein the crushed microbial cell product or a fraction thereof is mixed in a water slurry containing contaminants.

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