JP3533296B2 - Organic compound decomposition method - Google Patents

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention uses a microorganism.
The present invention relates to a method for decomposing organic compounds that decompose organic compounds .

[0002]

2. Description of the Related Art It has been known for a long time that microorganisms have the ability to oxidize and decompose various organic compounds, and they have long been represented by brewing, composting raw garbage and sludge, and water treatment. In addition, a technique for oxidizing and decomposing various organic compounds using microorganisms has been developed.

By the way, in recent years, pollution by harmful and hardly decomposable substances, particularly environmental pollution by substances represented by organic chlorine compounds has become a serious problem. For example, large amounts of organochlorine compounds have leaked into soil and groundwater around the sites of petroleum refineries, sites of various manufacturing plants, sites of cleaning plants, etc., resulting in the death of various organisms and abnormal reproduction. Is causing a large-scale environmental problem. Therefore, from the viewpoint of preserving biodiversity and maintaining the ecosystem, it is desired to prevent the spread of environmental pollution due to organochlorine compounds and to establish the technology for repairing and regenerating the polluted environment. Then, as a powerful means for treating harmful persistent substances, a technique utilizing microorganisms that oxidize and decompose organic compounds has been developed.

That is, as a conventional method for repairing and regenerating soil, groundwater, etc. contaminated with organic chlorine compounds,
Physicochemical methods such as vacuum extraction and activated carbon treatment after pumping groundwater have been tried. However, the physicochemical method has many problems such as low operability and low treatment efficiency for a wide range of low-concentration contamination but high cost. However, so-called bioremediation, which is a method for purifying soil and groundwater using microorganisms, has high treatment efficiency even for harmful persistent substances that are present in low concentrations in the environment, and the cost is low. However, since it has advantages such as a small load, it is expected to be a technology that can replace conventional physicochemical methods.

As a method of utilizing microorganisms for the treatment of harmful persistent substances, activated microorganisms that live in the environment such as soil or groundwater to be restored or regenerated are activated and persistent chlorine compounds such as organochlorine compounds Bios that oxidizes and decomposes substances to make them harmless
A method that uses timulation technology, a microorganism that oxidizes and decomposes harmful persistent substances such as organic chlorine compounds to render them harmless from the outside, and actively introduces them into the environment such as soil and groundwater.
Method using augmentation technology, and further, if necessary, transfer soil or groundwater contaminated with harmful persistent substances such as organic chlorine compounds from the landing, and use bioreactor etc. Oxidation of harmful and persistent substances
A method of disassembling has been tried and implemented. Also,
When using microorganisms to treat harmful persistent substances,
Methods for activating microorganisms include optimizing the growth environment of microorganisms and inducers of oxidases in order to increase the activity of oxidases required by microorganisms to oxidize and decompose target organic compounds. , Etc. have been studied and implemented.

For example, when the oxidase is an intracellular enzyme, in order to enhance the metabolic activity of the microorganism, temperature, pH,
For the purpose of optimizing the growth environment such as nutritional conditions, a medium containing nutrients necessary for the growth of microorganisms is added to the environment,
On the other hand, when the oxidase is an extracellular enzyme that oxidizes and decomposes the target organic compound by co-metabolization with each enzyme,
In this method, an inducer for releasing a large amount of the oxidase into the reaction system is searched, and the inducer is put into the environment by an optimum addition method.

[0007]

However, although the method of activating microorganisms by optimizing the entire environment can be applied to a closed system represented by bioreactor, activated sludge method, etc. When applied to microorganisms in a system, there is a problem that it is difficult to activate the microorganisms because it is difficult to completely optimize the entire living environment of the microorganisms.

Further, when a harmful persistent substance such as trichlorethylene (TCE) is oxidized and decomposed by using a microorganism, the decomposition of TCE by an aerobic microorganism is caused by the oxidation of methane oxidase, aromatic oxidase and the like. It is initiated by the initial oxidation of TCE by enzyme co-metabolism (core action), but the inducer that induces oxidases such as methane oxidase and aromatic oxidase is methane, phenol or toluene. Is a substance that has a detrimental effect on the environment, so addition of inducers that induce oxidases such as methane oxidase and aromatic oxidase to the environment causes secondary pollution to the environment and explosion accidents. There is a high possibility that methane oxidase, aromatic oxidase and other oxidase inducing substances cannot be added to the environment. It was.

Furthermore, even when a low-toxic natural organic compound is used as the inducer, COD in groundwater
Since there is a possibility of causing environmental damage such as increase in BOD and BOD, the use of a low-toxic natural organic compound as an inducer has a problem from the viewpoint of environmental protection.

Many species are known as microorganisms that oxidize and decompose hardly-decomposable substances such as organic chlorine compounds to render them harmless. However, as seen in the Bioaugmentation technology, these microorganisms are used in soil, Even if introduced directly into the environment such as groundwater, the number of introduced microorganisms rapidly decreases in the environment due to the competitive relationship with other microorganisms originally distributed in the soil, especially predation by protozoa. As a result, there is a problem in that it is not possible to achieve the expected effect when treating harmful persistent substances.

Further, in order to prevent the reduction of the microorganisms introduced into the environment, a large amount of a medium suitable for the growth of the administered microorganisms has been introduced into the environment. Since the living environment of the microorganisms introduced into the environment naturally depends on the introduced environment, it is difficult to optimize the environment for activating the microorganisms, and it is not economical, and the environment due to the addition of the medium is difficult. However, there was a problem that the second pollution occurs.

[0012] Further, after the microorganisms are loaded on the carrier, the microorganisms are introduced into the environment together with the carrier to protect the microorganisms introduced into the environment from competition with other microorganisms and predation, and to protect the microorganisms introduced into the environment. Attempts have been made to improve the survival rate and to oxidize and decompose persistently degradable substances such as organic chlorine compounds that are harmful to the environment for a long period of time to render them harmless. Although substances such as chlorine compounds are present in low concentrations, they are present in a wide range, so in order to decompose and detoxify substances such as organic chlorine compounds in the environment, a carrier supporting microorganisms should be used in the environment. However, there is a problem that it cannot be applied from the economical point of view and also from the viewpoint of microbial contamination.

The present invention has been made in view of the above-mentioned conventional examples, and can be applied not only to a closed system but also to an open system, and a microorganism can be used without administering a chemical substance such as an inducer for inducing an oxidase to an organic compound. It is an object of the present invention to provide a method for activating a microorganism capable of activating.

The present invention activates microorganisms not only in a closed system but also in an open system without administering a chemical substance such as an inducer for inducing an oxidase to an organic compound to activate an organic compound such as an organic chlorine compound. It is an object of the present invention to provide a method for decomposing an organic compound capable of efficiently oxidizing and decomposing a compound.

Furthermore, the present invention holds microorganisms at a high density and, when introduced into the environment, prevents the reduction of the microorganisms and efficiently oxidizes organic compounds such as organic chlorine compounds. It is an object of the present invention to provide a method for decomposing an organic compound capable of decomposing.

Further, the present invention activates microorganisms retained at a high density and, when the microorganisms are introduced into the environment, prevents the reduction of the microorganisms and efficiently organic compounds such as organic chlorine compounds. It is an object of the present invention to provide a method for decomposing an organic compound capable of oxidizing and decomposing.

Furthermore, the present invention activates microorganisms retained at a high density and, when the microorganisms are introduced into the environment, prevents the reduction of the microorganisms, and efficiently produces organic compounds such as organic chlorine compounds. It is an object of the present invention to provide a carrier that serves as a place for the oxidation and decomposition of

Further, the present invention is capable of activating a microorganism and oxidizing / degrading an organic compound such as an organic chlorine compound without administering an inducer for inducing an oxidase of an organic compound to the microorganism. It is an object of the present invention to provide a decomposition system for organic compounds that can be applied not only to closed systems but also to open systems.

[0019]

The method for activating a microorganism according to the first invention of the present application comprises a first metabolic system for carrying out an assimilation-type reduction reaction and a second metabolic system for carrying out an oxidation reaction. A substance serving as a substrate for the anabolic reduction reaction is administered to a microorganism to activate the first metabolic system, and the second metabolic system is activated based on the activation of the first metabolic system. The feature is to let.

According to the method for activating a microorganism in the first invention of the present application, an assimilation is performed on a microorganism having a first metabolic system for carrying out an assimilation-type reduction reaction and a second metabolic system for carrying out an oxidation reaction. A substance serving as a substrate for the type reduction reaction is administered to activate the first metabolic system. Next, the activated first metabolic system activates the second metabolic system.

The method for decomposing an organic compound according to the second invention of the present application comprises a first metabolic system for carrying out an assimilation-type reduction reaction and a second metabolic system for carrying out an oxidation reaction, Administration of a substance that serves as a substrate for a reduction reaction activates the first metabolic system, and a microorganism and an organic compound in which the second metabolic system is activated based on the activation of the first metabolic system. It is characterized in that the organic compound is decomposed by the microorganisms brought into contact with each other and the second metabolic system is activated.

According to the method for decomposing an organic compound in the second invention of the present application, an assimilation is carried out for a microorganism having a first metabolic system for carrying out an assimilation-type reduction reaction and a second metabolic system for carrying out an oxidation reaction. A substance that serves as a substrate for the type reduction reaction is administered,
The first metabolic system is activated, and the activated first metabolic system activates the second metabolic system. Then, the activated first metabolic system contacts the microorganism whose second metabolic system has been activated with the organic compound, and the organic compound is decomposed by the microorganism.

Further, in the method for decomposing an organic compound according to the third invention of the present application, a microorganism having a metabolic system for carrying out an oxidation reaction is carried on a carrier having a lipophilic polymer, and the carrier carrying the microorganism. And the organic compound are brought into contact with each other, and the organic compound is decomposed by the microorganism having the metabolic system.

According to the method for decomposing an organic compound in the third invention of the present application, a microorganism having a metabolic system for carrying out an oxidation reaction is supported on a carrier having a lipophilic polymer.
Then, the carrier supporting the microorganism is brought into contact with the organic compound, and the organic compound is decomposed by the microorganism.

Further, in the method for decomposing an organic compound according to the fourth aspect of the present invention, a microorganism having a metabolic system for carrying out an oxidation reaction is loaded on a carrier having a porous foam formed by foaming liquefied wood. Then, the carrier supporting the microorganism is brought into contact with the organic compound, and the organic compound is decomposed by the microorganism having the metabolic system.

According to the method for decomposing an organic compound in the fourth invention of the present application, a microorganism having a metabolic system for carrying out an oxidation reaction is supported on a carrier having a porous foam formed by foaming liquefied wood. It Then, the carrier supporting the microorganism is brought into contact with the organic compound, and the organic compound is decomposed by the microorganism.

Further, the carrier according to the fifth invention of the present application is characterized in that it comprises a porous foam formed by foaming liquefied wood.

According to the carrier of the fifth aspect of the present invention, the liquefied wood is foamed to form a porous porous foam having wood constituents.

Further, an organic compound decomposing system according to a sixth aspect of the present invention is a microorganism comprising a first metabolic system for carrying out an assimilation-type reduction reaction and a second metabolic system for carrying out an oxidation reaction, wherein the assimilation is carried out. It is characterized in that it is equipped with a mechanism for bringing into contact a substance and an organic compound which are substrates for the type reduction reaction.

According to the organic compound decomposition system of the sixth aspect of the present invention, a microorganism comprising a first metabolic system for carrying out an assimilation-type reduction reaction and a second metabolic system for carrying out an oxidation reaction, and assimilation-type reduction These three are brought into contact by the mechanism of bringing into contact the substance serving as a substrate of the reaction and the organic compound.

In the first, second and sixth inventions of the present application, the reaction system of the assimilation-type reduction reaction and the oxidation reaction is not limited as long as it is a system in which metabolism by the oxidation reaction is promoted by metabolism by the assimilation-type reduction reaction. There is no particular limitation.

In the first, second and sixth inventions of the present application, the assimilation-type reduction reaction means that the reduced pyridine nucleotide (NADH or NADPH), ferredoxin or the like is used as an electron donor to reduce the substrate and finally It is a reaction that produces a substance that is a raw material for a biosynthetic reaction.Metabolism systems that carry out assimilation-type reduction reactions include metabolism such as nitrogen fixation, assimilation-type nitrate reduction, assimilation-type nitrite reduction, and assimilation-type sulfate reduction The system can be mentioned. For example, in the assimilation-type nitrate reduction, nitrate ions are reduced to nitrite ions by the assimilation-type nitrate reductase, and the nitrite ions are finally reduced to ammonia by the assimilation-type nitrite reductase. At this time, reduced pyridine nucleotides (NADH or NADPH), ferredoxin, etc. are used as electron donors, and themselves shift to the oxidized form. The ammonia produced by the assimilative nitric acid reduction is used, for example, to produce the amino group of glutamic acid.

In the first, second and sixth inventions of the present application, the substance that serves as a substrate for the assimilation-type reduction reaction is not particularly limited as long as it can be a substrate that is required by the metabolic system of the microorganism. For example, various organic compounds, nitrate ions, sulfate ions, nitrite ions and the like can be mentioned. However, when an organic substance is used as a substrate when a substance that serves as a substrate for the assimilation-type reduction reaction is introduced into the environment such as soil or groundwater, the problem of secondary pollution due to the organic substance is likely to occur, as described above. The substrate is preferably an inorganic compound. Further, in consideration of the ease of handling and the like, it is more preferable to use a neutral salt. Furthermore, when a substance that serves as a substrate for the anabolic reduction reaction is administered to microorganisms, it is desirable to administer it in the form of an aqueous solution.

At this time, it is preferable that the concentration of the substance that serves as a substrate for the assimilation-type reduction reaction is a concentration that most activates the metabolic system of the microorganism to which it is administered. Since this concentration varies depending on the species of the microorganism, it should be appropriately determined depending on the target microorganism.However, in the case of nitrate, it is generally about 0.001 to 0.2 in the environment where the microorganism is present.
%, In the case of sulfate salts, in the presence of microorganisms
It is desirable to set it to about 0.001 to 0.1%.

In the first to fourth and sixth inventions of the present application, the oxidation reaction means dehydrogenation for removing electrons from the substrate, and the metabolic system for carrying out the oxidation reaction is pyruvin produced by aerobic bacteria. Examples thereof include metabolic systems such as acid utilization pathway and β-ketoadipate pathway that oxidizes aromatic compounds. For example, in the β-ketoadipic acid pathway, toluene is metabolized to β-ketoadipic acid via benzoic acid and catechol. In the metabolic system that executes the oxidation reaction, it was found that there is an internal pathway that uses reduced pyridine nucleotides (NADH or NADPH), ferredoxin, etc. as electron donors and transfers these to the oxidized form. ing.

In the first to fourth and sixth aspects of the present invention, the substance serving as a substrate for the oxidation reaction is not particularly limited as long as it is a substance required by the metabolic system of the microorganism and can serve as a substrate. Examples of the substance that serves as a substrate for such an oxidation reaction include malic acid, α-ketoglutaric acid, and various organic chlorine compounds. As various organic chlorine compounds, trichloroethylene, tetrachloroethylene, cis-dichloroethylene, trans-
Examples thereof include dichloroethylene, 1,1-dichloroethylene, monochloroethylene, vinyl chloride, and CB. In the first to fourth and sixth inventions of the present application, the enzyme that catalyzes the reaction to oxidize the substrate differs depending on the substrate, but when the substrate is an organic chlorine compound, for example,
Toluene oxidase, methane oxidase, ammonia oxidase and biphenyl oxidase are required.

In the first, second and sixth inventions of the present application, the microorganism is not particularly limited as long as it has both a metabolic system for executing the above-mentioned assimilation-type reduction reaction and oxidation reaction. As microorganisms, various bacteria,
Examples thereof include filamentous fungi or transforming fungi. Also,
In the third and fourth inventions of the present application, the microorganism is not particularly limited as long as it has both a metabolic system that executes the above-described oxidation reaction, and such microorganisms include various bacteria, filamentous fungi or A morphobacterium and the like can be mentioned. In addition, in the third and fourth inventions of the present application, the microorganism may perform the assimilation-type reduction reaction.

In the first to fourth and sixth inventions of the present application, when the substance serving as a substrate for the oxidation reaction is an organochlorine compound, the substance is an aerobic or facultative anaerobic bacterium, and the organochlorine compound, methane Various bacteria having a metabolic system that oxidizes, aromatic hydrocarbons, and ammonia can be preferably applied. Such bacteria usually have at least one enzyme selected from the group consisting of toluene oxidase, methane oxidase, ammonia oxidase and biphenyl oxidase. In the first to fourth and sixth inventions of the present application, among the bacteria specifically exemplified in Tables 1 and 2, the species satisfying the above requirements can be particularly preferably used.

[0039]

[Table 1]

[Table 2] Incidentally, in the first to fourth and sixth inventions of the present application, as the microorganisms, those already isolated, those newly screened from the soil or the like according to the purpose can be appropriately used, and a mixture of a plurality of types of microorganisms can be used. It may be a system. However,
When the substance used as the substrate for the oxidation reaction is an organochlorine compound, YMCT-001 strain (reposited by the Institute of Biotechnology, Institute of Biotechnology, FERM BP-52, which has nitrate reduction ability as a microorganism)
The application of 82) is more preferable because the oxidation / decomposition of the organochlorine compound is promptly executed.

Here, in the first, second and sixth inventions of the present application, a supplementary explanation will be given on the mechanism by which microorganisms are activated.

In the first, second and sixth inventions of the present application, it is not required that the enzyme itself which catalyzes the oxidation reaction is induced in the microorganism, and the metabolic system for carrying out the anabolic reduction reaction possessed by the microorganism is provided. It is characterized by activating a metabolic system that carries out an oxidative reaction of a microorganism by administering a activating substance to the microorganism.

Conventionally, attempts have been made to promote the oxidation reaction by adding an inorganic compound such as hydrogen peroxide or potassium permanganate to the field of the oxidation reaction of an organic compound by a microorganism. The method of adding an inorganic compound such as manganese potassium activates the metabolic system of the microorganism and does not oxidize or decompose the substrate by the metabolic system of the microorganism, but the oxidizing power of the inorganic compound such as hydrogen peroxide or potassium permanganate. Oxidize the substrate directly by the above, and even if the oxidation / degradation of the substrate is promoted, it does not result from acting on the metabolic system of the microorganism to activate this metabolic system.

The present inventors, when utilizing the oxidation reaction of an organic compound by a microorganism, by administering to the microorganism an inorganic compound that activates a metabolic system possessed by the microorganism for executing the assimilation-type reduction reaction, It was found that the metabolic system possessed by the microorganisms, which carries out the oxidation reaction, is activated, and the present invention has been completed.

At present, by administering to a microorganism an inorganic compound that activates a metabolic system that carries out an anabolic reduction reaction,
The details of the mechanism of activation of the metabolic system that the microorganism carries out the oxidation reaction have not been clarified, and it is under intensive study, but from the findings to date, the inventors have shown the following: I am considering.

That is, 1. Generated by a metabolic system that executes an anabolic reduction reaction because a reaction in which a oxidase oxidizes a substrate in a metabolic system that performs an oxidative reaction of a microorganism and a metabolic system that executes an anabolic reduction reaction of a microorganism are simultaneously driven. The resulting oxidizing power is utilized in the metabolic system that executes the oxidation reaction, and the metabolic system that executes the oxidation reaction of the microorganism is activated.

2. As described above, reduced pyridine nucleotides (NADH or NADPH) are consumed in metabolic systems that carry out oxidative reactions of microorganisms, while reduced pyridine nucleotides are also used in metabolic systems that carry out anabolic reduction reactions. Is consumed, the biosynthesis rate of reduced pyridine nucleotides is increased by activating the metabolic system that carries out anabolic reduction reactions, and a large amount of reduced pyridine nucleotides are produced. The metabolic system, which carries out the oxidative reaction of the microorganisms with consumption, is activated.

Based on the mechanism described above, it is speculated that when an inorganic compound that activates a metabolic system that carries out an anabolic reduction reaction is administered to a microorganism, the metabolic system that the microorganism has and carries out an oxidation reaction is activated. ing.

In the second invention of the present application, the method of contacting the organic compound with the microorganism in which the metabolic system that executes the oxidation reaction is activated by activating the metabolic system that executes the assimilation-type reduction reaction is not particularly limited. It is not done. As such a method, a method using a bioreactor, a method of directly contacting a microorganism with an organic compound in the environment, or the like can be used. When using a bioreactor,
A pre-activated microorganism and an organic compound may be brought into contact with each other in the bioreactor, or a microorganism and an organic compound may be brought into contact with each other in the bioreactor in advance, and then a metabolic system for executing an assimilation-type reduction reaction in the bioreactor. You may make it add the substance which activates. However, in order to promptly induce the oxidation reaction, it is preferable to bring the previously activated microorganism and the organic compound into contact with each other in the bioreactor. The environment inside the bioreactor is maintained at a state where the reaction rate of the oxidation reaction by microorganisms is the highest by adding substances that activate the metabolic system that executes anabolic reduction reaction into the bioreactor as necessary. To be done. When using a method of directly contacting a microorganism with an organic compound in the environment, a microorganism originally inhabiting the environment may be activated to bring the microorganism into contact with the organic compound in the environment. Alternatively, a microorganism known to oxidize and decompose an organic compound in the environment may be previously cultured, and the cultured microorganism may be added to the environment. When the cultured microorganisms are put into the environment, it is preferable to put the previously activated microorganisms into the environment as described above. In addition, if necessary, the environment in which the microorganisms were introduced has a high reaction rate of the oxidation reaction by the microorganisms, for example, by adding a substance that activates the metabolic system that executes the anabolic reduction reaction into the environment. Try to keep it as it is.

In addition, it is preferable to support the microorganisms on the carrier, whichever method is used, that is, a method using a bioreactor or a method of directly contacting the microorganisms with an organic compound in the environment. By supporting the microorganisms on a carrier, it becomes possible to maintain the microorganisms at a high density and protect the microorganisms from the competition for survival with other organisms, so the oxidation and decomposition of organic compounds should be carried out efficiently. You can Such a carrier can be appropriately changed depending on the microorganism species, but by using the carrier described in the third, fourth and fifth inventions of the present application, the oxidation / decomposition of the organic compound can be carried out more efficiently. Therefore, it is preferable.

In the second invention of the present application, the organic compound to be brought into contact with the microorganism may be in the form of the organic compound itself, dissolved in a solvent, or present in soil, water or air. At this time, when an organic compound such as an organic chlorine compound is in the form of existing in soil, water or air, the soil activated by the above activation method is contaminated with harmful persistent substances. The soil and the like can be purified by the microorganisms by bringing them into contact with the like. The term "soil" is a concept that includes all of the environment below the surface of the earth, and refers to soil, soil water, groundwater, and all that include them.

When treating soil contaminated with organic chlorine compounds, etc., the excavated contaminated soil is sized to form a slurry, and as described above, the microorganisms activated using the bioreactor and the contaminated soil are separated. A method of contacting and oxidizing and decomposing organic chlorine compounds, etc., or pumping groundwater contaminated with organic chlorine compounds, etc. and contacting the microorganisms activated in the bioreactor with groundwater to oxidize organic chlorine compounds, etc. It is possible to use a method of decomposing or introducing a microorganism and a substance that activates a metabolic system that carries out anabolic reduction reaction into the soil, and oxidizing and decomposing organic chlorine compounds etc. in situ. it can. A substance that activates the metabolic system that executes the assimilation-type reduction reaction can minimize the loss of time and energy due to stirring work, etc., compared to the case of administering it to a microorganism in a solid state. It is desirable to contact the microorganism in form.

Further, in the third invention of the present application, the carrier holding the microorganism is brought into contact with the organic compound, but if the microorganism is held in the carrier, the degree of accumulation of the microorganism is improved and the microorganism is kept in soil or the like. When introduced into the environment of, it is possible to protect the microorganisms introduced into the environment from survival competition and predation with the microorganisms that originally live in the environment, improving the survival of the microorganisms in the environment, It becomes possible to maintain the oxidation / decomposability of organic compounds for a long period of time.

Further, in the third invention of the present application, since the lipophilic polymer is used as the carrier for holding the microorganism, it is possible to hold and concentrate the organic compound such as the organic chlorine compound in the vicinity of the microorganism. .

Here, the lipophilic polymer is a polymer having the ability to absorb and retain an oily substance several times to several tens of times its own weight. The lipophilic polymer is a polymer having a lipophilic group having a high affinity for an oily substance and having a low cross-linking degree using a lipophilic monomer as a basic unit.

As the lipophilic polymer used in the third invention of the present application, a polymer having high gel strength and excellent salt resistance is preferable in consideration of the use environment in which it is used by being put into the environment such as soil. For example, an acrylic acid-based polymer such as a highly oil-absorbing polymer or a norbornene ring-opening polymer may be mentioned.
These can be used alone or in combination of two or more. Lipophilic polymers are powdery, powdery,
Various forms such as fine powder, pearl, beads, flakes or blocks can be used.
At this time, the particle size of the lipophilic polymer is 0.1 μm.
It is preferably about several mm. The particle size is defined as the diameter of the smallest sphere that can contain the polymer therein.

The carrier used in the third invention of the present application may include a porous body capable of retaining microorganisms.
As such a porous body, for example, one capable of forming a microhabitat of microorganisms is preferable. The micro habitat is a minute residence of a microorganism in a pore of about several μm and has a function of protecting the microorganism from a harsh environment. For example, even if the external environment is in a dry state that affects the survival of the microorganisms, the water supply to the microorganisms is maintained because the capillary water is retained in the microhabitat. In addition, the microorganisms in the microhabitat are protected from the predation of protozoa in the environment such as soil. Therefore, it becomes possible to improve the viability of microorganisms by artificially forming a microhabitat with a carrier containing a porous body.

The porous body can be selected in various forms such as granular or layered form. Examples thereof include soil particles having an aggregate structure such as ceramics, glass, calcium silicate, silica, alumina and Kanuma soil. It is possible to use one kind or a combination of two or more kinds of porous materials made of inorganic materials and organic materials such as activated carbon, urethane foam, photocurable resin, anion exchange resin, cellulose, lignin, chitin and chitosan. As these porous bodies, inexpensive ones are desirable. Further, in consideration of the use environment in which it is also used by being used in the environment such as soil, it is desirable that the dispersibility, mobility, etc. in the environment such as soil is not impaired. Particles of about mm are preferable. Further, it is desirable to have a structure suitable for retaining and growing microorganisms, for example, several μm to several tens μ.
A porous body having a pore size of m is desirable.

As a method for producing a carrier containing a porous body, for example, fine powder of the lipophilic polymer and the porous body may be mixed and gently stirred to be mixed. When a lipophilic polymer and a porous body are mixed and put into an environment such as soil, in many cases, the porous body keeps contact with the environment such as soil and the lipophilic polymer, depending on the feeding conditions. Both are in contact with each other.

Further, when it is required to bring the lipophilic polymer and the porous body closer to each other more strictly, for example, by mixing fine powder of the lipophilic polymer and the porous body and heating the both, Can be used, or when the porous body is urethane foam, a method of mixing urethane with fine powder of a lipophilic resin and then foaming urethane can be used. Then, the microorganisms can be introduced into the pores existing in the porous body by adding a suspension of the microorganisms to these carriers and subjecting them to a decompression treatment.

In general, organic compounds such as organic chlorine compounds are present in a low concentration over a wide range in the environment such as soil, especially in groundwater in a saturated layer. According to the third invention of the present application, the lipophilic polymer makes it possible to absorb and retain an organic compound such as an organic chlorine compound in the vicinity of a microorganism even in soil. Therefore, the contact time between the microorganism and the organic compound such as the organic chlorine compound becomes long, and the organic compound can be efficiently oxidized and decomposed.

That is, according to the third invention of the present application, unlike the conventional bioremediation method, unlike the method of collecting microorganisms in the vicinity of the organic compound diffused in the environment such as soil, the absorption of the lipophilic polymer is carried out. The organic compound can be concentrated in the vicinity of the microorganism by moving the organic compound in the environment using force as a power. In addition, when organic compounds such as organochlorine compounds are partially present in high concentrations, the organic compounds are absorbed and retained in the lipophilic polymer and gradually released, so that the microorganisms become high-concentration organic compounds. It is possible to avoid the death of microorganisms, the decrease in activity, etc. due to the contact. In addition, some microorganisms that oxidize and decompose organic compounds such as organic chlorine compounds require the coexistence of a substance called an inducer in order to induce an enzyme that acts as a catalyst in the metabolic system. For example, a group of bacteria known as trichlorethylene-decomposing bacteria often requires an aromatic hydrocarbon compound such as phenol or guar ruene as an inducer when decomposing trichlorethylene. In this case, in order to maintain the oxidation / decomposition of the organic compound such as the organic chlorine compound, not only the microorganism remains, but the inducer is maintained in the vicinity of the microorganism, for example, in the environment such as soil for a long time, and , It is necessary to continue to be supplied.

The indexer is often used as a nutrient source for microorganisms that are not particularly involved in the desired oxidation reaction, for example, microorganisms that naturally inhabit the soil. It is also difficult to make it exist locally in the vicinity. Therefore, when the lipophilic polymer is impregnated with the inducer, the inducer is constantly supplied only to the microorganisms that carry out the desired oxidation reaction, so that the oxidation / degradability of organic compounds such as organochlorine compounds by the microorganisms is maintained. .

Generally, lipophilic polymers are mainly used for the purpose of removing harmful substances such as organic chlorine compounds from water. However, such usage is
It simply moves harmful substances such as organochlorine compounds from the target such as water. In particular, when cleaning soil contaminated with harmful substances such as organochlorine compounds, even if the lipophilic polymer is put into the soil to absorb harmful substances such as organochlorine compounds, the lipophilic polymer is absorbed. If harmful substances such as organochlorine compounds are not oxidized or decomposed in the soil, the lipophilic polymer must be recovered again. In reality, it is difficult to recover the lipophilic polymer from the soil. It does not fundamentally solve the demand for oxidizing and decomposing harmful substances such as chlorine compounds in soil.

The third invention of the present application is generally applicable to any microbial species, and the microbial species, lipophilic polymer and porous body are appropriately selected in accordance with the organic compound to be oxidized and decomposed. can do. Further, as a method of contacting an organic compound with a carrier holding a microorganism in which a metabolic system that executes an oxidation reaction is activated by activating a metabolic system that executes an assimilation-type reduction reaction, the second invention of the present application In addition, the form of the organic compound to be brought into contact with the microorganism may be the organic compound itself, may be in a state of being dissolved in a solvent, or may be a form existing in soil, water or the atmosphere. Good. Further, when treating soil contaminated with an organic chlorine compound or the like, the method described in the second aspect of the present invention can be used.

By using the third invention of the present application in an environment such as soil, it is possible to oxidize and decompose organic compounds existing in the environment. When the third invention of the present application is used in an environment such as soil, the method of introducing the carrier holding the microorganisms into the environment can be carried out by a conventional method such as spraying treatment or mixing treatment with soil or the like. As the method of administration to the saturated layer, a method of providing an injection well and introducing / dispersing the carrier holding the microorganisms therein, a method of suspending in water and adding / dispersing the suspension, and the like are selected. be able to.

In the fourth invention of the present application, the carrier holding the microorganisms is brought into contact with the organic compound. However, as described above, holding the microorganisms in the carrier improves the degree of accumulation of the microorganisms, and When introduced into the environment such as soil, the microorganisms introduced into the environment can be protected from survival competition with microorganisms that originally live in the environment and predation, and the survival of the microorganisms in the environment It is possible to improve the temperature and maintain the oxidation / decomposition ability of organic compounds for a long time.

Further, in the fourth invention of the present application, since a carrier having a porous foam formed by foaming liquefied wood is used as the carrier, a substance such as an aromatic compound is further added from the outside. It is possible to supply the assimilatory substance to the microorganisms without using it. Therefore, when the present invention is carried out in the environment, it is possible to supply the assimilable substance to the microorganisms without introducing a large amount of the aromatic compound into the environment. It is also possible to protect the introduced microorganisms from survival competition and predation with microorganisms that originally live in the environment, improve survival of microorganisms in the environment, and maintain the oxidation and degradability of organic compounds for a long time. It becomes possible.

In the fourth and fifth inventions of the present application, the porous foam formed by foaming liquefied wood means that wood and polyhydric alcohol such as caprolactone polyester polyol are heated together with a liquefaction catalyst such as sulfuric acid. Isocyanate (MDI) was added to the liquefied wood obtained, and a foaming agent,
As a neutralizing agent, a catalyst and a foam stabilizer, water, imidazole, amine-based substances such as tetramethylhexamethylenediamine and pentamethyldiethylenediamine, and silicone-based substances such as silicone polymers are applied and foamed to form a porous structure. It is a pawn. Therefore, the constituents of the porous body include a large number of natural organic compounds derived from wood or having an aromatic system or a double bond, and when microorganisms that assimilate aromatic compounds are retained on this carrier. Is
Since the above-mentioned natural organic compound is gradually eluted from the carrier itself, the assimilation substance is supplied to the microorganism. as a result,
Microorganisms are always supplied with assimilating substances, and when introduced into the environment, they can protect the microorganisms from the competition for survival with the microorganisms that originally live in the environment and predation. It is possible to improve the property and maintain the oxidation / decomposability of organic compounds for a long period of time.

Since the porous foam formed by foaming liquefied wood has high biodegradability, even when it is thrown into the environment such as soil or ground water, it accumulates in the environment and causes secondary pollution. Very unlikely. Further, the porous foam formed by foaming the liquefied wood can have a form as required, but the pore diameter is from several μm to a maximum of several hundred μm.
Therefore, it becomes possible to retain the microorganism at a higher concentration. Furthermore, the carrier provided with the porous foam formed by foaming liquefied wood can form the mikrohabitat of microorganisms, and thus it becomes possible to improve the viability of microorganisms in the environment. Of.

Further, the carrier may be formed by combining a porous foam formed by foaming liquefied wood with another material. As the other material, various forms such as granular or layered can be selected. For example, ceramics,
Inorganic materials such as glass, calcium silicate, silica, alumina and soil particles with aggregate structure such as Kanuma soil, activated carbon, urethane foam, photocurable resin, anion exchange resin, cellulose, lignin, chitin and chitosan It is possible to use one kind or two or more kinds of porous materials made of materials in combination. As these porous bodies, inexpensive ones are desirable. Further, in consideration of the use environment in which it is also used by being used in the environment such as soil, it is desirable that the dispersibility, mobility, etc. in the environment such as soil is not impaired. Particles of about mm are preferable. Further, it is desirable to have a structure suitable for retaining and growing microorganisms, for example, several μm to several tens μ.
A porous body having a pore size of m is desirable.

As a method for producing a carrier compounded with another substance, when another substance is combined with an inorganic porous material or an organic porous material, for example, a porous foam made of liquefied wood of an appropriate size is used. The body and other materials are mixed in an appropriate ratio and heated with stirring to heat-bond. Then, a microorganism can be introduced into the pores existing in the carrier by adding a suspension of the microorganism to this carrier and subjecting it to a pressure reduction treatment. When the other substance is a hydrophilic gel, for example, the hydrophilic gel in which microorganisms are suspended may be impregnated in a porous foam made of liquefied wood and then solidified.

Further, when an inorganic porous body or an organic porous body is used as another substance, the porous foam made of liquefied wood and the inorganic porous body or the organic porous body are more strictly integrated. When it is required to do so, when the liquefied wood is foamed, an appropriate amount of inorganic porous material or organic porous material is added and gently stirred and then foamed. In this way, it becomes possible to form a layer of a porous foam made of liquefied wood on the surface of the inorganic porous material or the organic porous material. Then, as described above, the microorganisms can be introduced into the pores existing in the carrier by adding a suspension of the microorganisms to this carrier and subjecting it to a decompression treatment.

In the fourth and fifth inventions of the present application, when bringing the groundwater pumped up and the excavated soil into contact with the carrier, the particle size of the carrier should be taken into consideration in terms of the efficiency of oxidation and decomposition of organic compounds. And decide. However, when it is directly put into the environment with soil, it is desirable that it does not impair the movement and dispersibility in the environment, for example, several hundred μm
It is desirable that the particle size be about several mm.

In the fourth invention of the present application, as the microorganisms, those already isolated and those newly screened according to the purpose can be appropriately used, and a mixed system of microorganisms composed of a plurality of species can be used. In the fourth invention of the present application, a carrier having a porous foam formed by foaming liquefied wood that retains microorganisms is directly placed in an environment such as soil or groundwater containing an organic compound such as an organic chlorine compound. Purification of the environment such as soil and groundwater by oxidizing and decomposing organic compounds such as organochlorine compounds without throwing in large amounts of substances that have a high impact on the environment such as phenol and toluene by throwing in or contacting them. Can be carried out.

The fourth invention of the present application is generally applicable to any microbial species, and the microbial species and carrier can be appropriately selected according to the organic compound to be oxidized / decomposed. Further, as a method of contacting an organic compound with a carrier holding a microorganism in which a metabolic system that executes an oxidation reaction is activated by activating a metabolic system that executes an assimilation-type reduction reaction, the second invention of the present application The method described in 1 can be used, and as the form of the organic compound to be brought into contact with the microorganism, the organic compound itself may be used,
It may be in a state of being dissolved in a solvent, or may be in a form existing in soil, water or the atmosphere. Further, when treating soil contaminated with an organic chlorine compound or the like, the method described in the second aspect of the present invention can be used. Also,
When bringing the carrier into contact with groundwater, a method such as adding the carrier to the pumped groundwater can be used, but if the carrier is added / suspended to the pumped groundwater and then recirculated into the groundwater again, , -Soil and groundwater can be treated every time, which is preferable.

The sixth invention of the present application is a microorganism comprising a first metabolic system for carrying out an assimilation-type reduction reaction and a second metabolic system for carrying out an oxidation reaction, a substance serving as a substrate for the assimilation-type reduction reaction, and an organic substance. It has a mechanism for bringing the compounds into contact.

In the sixth invention of the present application, a microorganism comprising a first metabolic system for carrying out an assimilation-type reduction reaction and a second metabolic system for carrying out an oxidation reaction, a substance serving as a substrate for the assimilation-type reduction reaction, and an organic substance The mechanism for bringing the compound into contact is not particularly limited as long as it can realize the contact of these three members. As such a mechanism, a bioreactor or a mechanism for directly contacting the three in an environment can be used.

When a bioreactor is applied, the microorganism previously activated with a substance serving as a substrate for the assimilation-type reduction reaction may be brought into contact with the organic compound in the bioreactor, or the microorganism and the organic compound may be contacted with each other. After contacting in advance in the reactor, a substance that activates the metabolic system that executes the assimilation-type reduction reaction may be added into the bioreactor. However, in order to promptly induce the oxidation reaction, it is preferable to bring the previously activated microorganism and the organic compound into contact with each other in the bioreactor.

Regarding the environment in the bioreactor, the reaction rate of the oxidation reaction by the microorganism is maximized by, for example, adding a substance that activates the metabolic system that executes the assimilation-type reduction reaction into the bioreactor. Maintained in a state.
When using a method of directly contacting a microorganism with an organic compound in the environment, a microorganism that originally inhabits the environment is activated by administering a substance that activates a metabolic system that carries out an anabolic reduction reaction. , This microorganism may be brought into contact with an organic compound in the environment, or a microorganism known to oxidize and decompose an organic compound in the environment may be pre-cultured and assimilated with the microorganism cultured in the environment. A substance that activates the metabolic system that executes the type reduction reaction may be added. When the microorganisms cultured in the environment are introduced, it is preferable to introduce the microorganisms previously activated by a substance that activates the metabolic system that executes the assimilation-type reduction reaction into the environment as described above. In addition, if necessary, a substance that activates the metabolic system that executes the assimilation-type reduction reaction may be added to the environment at any time to keep the three in contact with each other, so that the reaction rate of the oxidative reaction by microorganisms may be small. Try to keep it high.

As the mechanism for bringing the three into contact with each other, whichever mechanism is used, which is in direct contact with a bioreactor or a microorganism, an organic compound and a substance that activates a metabolic system that carries out the assimilation-type reduction reaction in the environment is used. Also, it is preferable to support the microorganisms on the carrier. By supporting the microorganisms on a carrier, it becomes possible to maintain the microorganisms at a high density and protect the microorganisms from the competition for survival with other organisms, so the oxidation and decomposition of organic compounds should be carried out efficiently. You can Such a carrier can be appropriately changed depending on the microorganism species, but by using the carrier described in the third, fourth and fifth inventions of the present application, the oxidation / decomposition of the organic compound can be carried out more efficiently. Therefore, it is preferable.

In the sixth invention of the present application, the organic compound to be contacted with the microorganism may be in the form of the organic compound itself, dissolved in a solvent, or present in soil, water or air. At this time, when the organic compound such as an organic chlorine compound is in a form existing in soil, water or air, it is activated by contact with a substance that activates a metabolic system that executes an assimilation-type reduction reaction. By contacting the microorganisms with the soil or the like contaminated with harmful persistent substances, the microorganisms can purify the soil or the like.

When treating soil contaminated with organochlorine compounds, etc., the excavated contaminated soil is sized to form a slurry, and as described above, the three are brought into contact with each other by the bioreactor to remove the organochlorine compounds and the like. A method of oxidizing and decomposing, pumping groundwater contaminated with organochlorine compounds, etc., while activating the microorganism by contacting the substance that activates the metabolic system that executes the assimilation-type reduction reaction in the bioreactor with the microorganism In the soil, a method of oxidizing and decomposing organic chlorine compounds or the like by contacting these or a microorganism, and a substance that activates a metabolic system that executes an assimilation-type reduction reaction are introduced into the soil,
A method of oxidizing and decomposing an organic chlorine compound or the like by bringing the three into contact with each other in situ can be used. A substance that activates the metabolic system that executes the assimilation-type reduction reaction can minimize the loss of time and energy due to stirring work, etc., compared to the case of administering it to a microorganism in a solid state. It is desirable to contact the microorganism in form.

All the first to sixth inventions of the present application are
It can be preferably used for the oxidation / decomposition of environmental pollutants typified by organic chlorine compounds, etc., but is also applied to systems that produce useful substances by oxidizing / decomposing other organic compounds as necessary. It goes without saying that it is possible.

[0084]

BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of the present invention will be described in detail below with reference to the drawings. In addition, in each drawing, the same components are denoted by the same reference numerals, and overlapping description for each drawing is omitted.

An example of the constitution of an organic compound decomposing system according to the present invention is shown in FIGS.

In FIG. 1, 1 is a pump for pumping groundwater from the groundwater layer 2 to the ground, and 3 is the groundwater layer 2.
A pumping well 4 that serves as a flow path for pumping more groundwater, an addition device 4 for adding to the groundwater pumped through the pumping well 3 a substance that activates a metabolic system that carries out an anabolic reduction reaction of microorganisms, and a 5 Introducing device for introducing a predetermined microorganism into groundwater pumped through the pumping well 3, 6 is an adding device 4
And an injection well that recirculates into the environment groundwater in which the substance that activates the metabolic system that carries out the assimilation-type reduction reaction of the microorganism and the microorganism is administered by the introduction device 5, and 7
It is a connecting pipe that connects the pumping well 3 and the injection well 6.
In addition, 8 and 9 have shown the groundwater level and the impermeable layer, respectively.

In the organic compound decomposition system shown in FIG. 1, groundwater is pumped from the groundwater layer 2 through the pumping well 3 by using the pumping pump 1 as a driving force. The pumped-up groundwater passes through an addition device 4 and an introduction device 5 through a connecting pipe 7, and in the process, a substance that activates a metabolic system that carries out an assimilation-type reduction reaction of microorganisms and microorganisms are added to the groundwater. Then, the groundwater into which the substance that activates the metabolic system that carries out the assimilation-type reduction reaction of microorganisms and the microorganisms are input is returned to the environment through the injection well 6.

The microorganisms held in the introduction device 6 can be activated in advance with a substance that activates the metabolic system that executes the assimilation-type reduction reaction. Further, the addition device 4 and the introduction device 5 operate independently of each other, and if necessary, an optimal amount of a substance that activates a metabolic system that executes the assimilation-type reduction reaction and a microorganism are added.

Further, in FIG. 2, 10 is a bioreactor which serves as a reaction site for the oxidation / decomposition of an organic compound by a microorganism, and 11 is a bioreactor 10 and activates a metabolic system for carrying out anabolic reduction reaction of the microorganism. An addition device for supplying a substance to be supplied, 12 is an introduction pipe for introducing groundwater pumped from the groundwater layer 2 into the bioreactor 10 through the pumping well 3, and 13 is injection of groundwater in which the oxidation / decomposition of the organic compound is completed by the bioreactor 10. It is a return pipe for returning to the well 6.

In the organic compound decomposition system shown in FIG. 2, groundwater is pumped from the groundwater layer 2 through the pumping well 3 using the pumping pump 1 as a driving force. The pumped ground water is supplied to the bioreactor 10 through the introduction pipe 12. The bioreactor 10 is appropriately supplied with a substance that activates the metabolic system that executes the assimilation-type reduction reaction of microorganisms from the addition device 11, and the activated microorganisms can oxidize and decompose organic compounds in groundwater. To be executed.
Then, the groundwater in which the oxidation / decomposition of the organic compound has been performed inside the bioreactor 10 is injected into the injection well 6 through the reflux pipe.
To the environment through the injection well 6.

In the decomposition system of the organic compounds shown in FIGS. 1 and 2, the morphology of the microorganisms is a solid (including a slurry) supported on a carrier, a lyophilized powder, a mixture thereof, or the microorganism itself. A turbid suspension or the like can be selected according to the topography and purpose of the environment where the system is installed. Further, in the present system, by adopting the above-mentioned mechanism, the microorganism activates the metabolic system that executes the assimilation-type reduction reaction, and efficiently oxidizes and decomposes organic compounds without imposing an environmental burden. It becomes possible to execute.

Next, a preferred embodiment of the present invention will be described based on experimental results.

Example 1 In 0.1M phosphate buffer,
After dissolving sodium nitrate so that the final concentration was 0.015M, 25 ml of the solution was stored in a vial. Next, 25 ml of YMCT-001 strain having an OD 660 of about 1 was suspended in this solution, trichlorethylene (TCE) was added to 20 ppm, and the vial was sealed with a butyl rubber septum. From the results of various redox tests, it has been clarified that the YMCT-001 strain reduces nitric acid.

Then, the YMCT-001 strain was shake-cultured by shaking the vial at 25 ° C. and 100 rpm, and the TCE concentration in the vial was measured with time. The TCE concentration was measured by the headspace method by measuring the TCE concentration in the gas phase with a PID (photoionization detector) using a gas chromatograph. The results are shown in FIG.

When the concentration of nitrate ion in the solution in the vial was measured at the end of the test, it was lowered to 9.6 × 10 ^-4 M.

Example 2 A test was conducted under exactly the same conditions as in Example 1 except that cis-dichloroethylene was added in place of trichloroethylene so that the concentration was 1 ppm.

Table 3 shows the residual concentration of cis-dichloroethylene after 4 days from the start of the test.

Example 3 Except that trans-dichloroethylene was added instead of cis-dichloroethylene.
The test was conducted under the same conditions as in Example 2.

Table 3 shows the residual concentration of trans-dichloroethylene after 4 days from the start of the test.

Example 4 A test was conducted under exactly the same conditions as in Example 2 except that 1,1-dichloroethylene was added instead of cis-dichloroethylene.

Table 3 shows the residual concentration of 1,1-dichloroethylene after 4 days from the start of the test.

Example 5 Example 2 was repeated except that monochloroethylene was added instead of cis-dichloroethylene.
The test was conducted under exactly the same conditions as above.

Table 3 shows the remaining monochloroethylene concentration after 4 days from the start of the test. (Example 6) Example 2 was repeated except that PCB (Kanegafuchi Chemical Co., Ltd .: Kanechlor KC-300) was added instead of cis-dichloroethylene.
The test was conducted under exactly the same conditions as above.

In Table 3, P after 4 days from the start of the test
The remaining concentration of CB is shown.

Example 7 A test was conducted under exactly the same conditions as in Example 2 except that tetrachloroethylene was added instead of cis-dichloroethylene.

Table 3 shows the residual concentration of tetrachlorethylene after 4 days from the start of the test.

Example 8 A test was conducted under exactly the same conditions as in Example 1 except that iron sulfate was added instead of sodium nitrate. A similar test was conducted. From the results of various oxidation-reduction tests, it has been clarified that the YMCT-001 strain carries out a slight sulfuric acid reduction.

The results are shown in FIG.

(Example 9) After putting the soil in the entire container having an internal volume of about 0.4 m ³ , TCE was added so that the TCE concentration in the soil was about 1 ppm on average. Next, sodium nitrate was dissolved in 0.1M phosphate buffer so that the final concentration would be 0.015M, and OD 660 reached about 1 in 100 l of this solution.
After suspending 50 liters of CT-001 strain, this suspension was added to the soil. Then, while keeping this container at 25 ° C., the TCE concentration in the container was measured with time in the same manner as in Example 1.

The results are shown in FIG.

Comparative Example 1 A test was conducted under the same conditions as in Example 1 except that ammonium phosphate was added instead of sodium nitrate.

The results are shown in FIG.

At the end of the test, the ammonium ion concentration in the solution in the vial was measured and found to be 0.015M.
Therefore, no change in the concentration was detected as compared with the start of the test.

(Comparative Example 2) A test was carried out under the same conditions as in Comparative Example 1 except that cis-dichloroethylene was added so as to be 1 ppm instead of trichlorethylene.

Table 3 shows the residual concentration of cis-dichloroethylene 4 days after the start of the test.

COMPARATIVE EXAMPLE 3 Except that trans-dichloroethylene was added instead of cis-dichloroethylene.
The test was performed under the same conditions as in Comparative Example 2.

Table 3 shows the residual concentration of trans-dichloroethylene after 4 days from the start of the test.

Comparative Example 4 A test was conducted under the same conditions as in Comparative Example 2 except that 1,1-dichloroethylene was added instead of cis-dichloroethylene.

Table 3 shows the residual concentration of 1,1-dichloroethylene after 4 days from the start of the test.

Comparative Example 5 Comparative Example 2 except that monochloroethylene was added instead of cis-dichloroethylene.
The test was conducted under exactly the same conditions as above.

Table 3 shows the residual monochloroethylene concentration after 4 days from the start of the test. (Comparative Example 6) Comparative Example 2 except that PCB (Kanegafuchi Chemical Co., Ltd .: Kanechlor KC-300) was added instead of cis-dichloroethylene.
The test was conducted under exactly the same conditions as above.

In Table 3, P after 4 days from the start of the test
The remaining concentration of CB is shown.

(Comparative Example 7) A test was conducted under the same conditions as in Comparative Example 2 except that tetrachloroethylene was added instead of cis-dichloroethylene.

Table 3 shows the residual concentration of tetrachloroethylene after 4 days from the start of the test.

[0125]

[Table 3] (Comparative Example 8) In Example 9, an equal amount of the suspension solution was used.
The TCE concentration in the container was measured with time in the same manner as in Example 9 except that 0.1M phosphate buffer was used.

The results are shown in FIG.

Comparative Example 9 The TCE concentration in the container was measured with time in the same manner as in Example 9 except that ammonium phosphate was added instead of sodium nitrate.

The results are shown in FIG.

As is clear from Examples 1 and 8 and Comparative Example 1, oxidation of an organochlorine compound in a microorganism is carried out by bringing the microorganism into contact with a substance that activates a metabolic system that carries out an anabolic reduction reaction of the microorganism.・ It can be understood that the decomposition is promoted and the organic chlorine compounds are efficiently decomposed.

As is clear from Examples 2 to 7 and Comparative Examples 2 to 7, by contacting a microorganism with a substance that activates the metabolic system that carries out the assimilation-type reduction reaction of the microorganism, a substance other than TCE It can be understood that many organochlorine compounds are efficiently decomposed. Further, Example 9,
As is clear from Comparative Examples 8 and 9, by contacting a microorganism with a substance that activates a metabolic system that carries out an anabolic reduction reaction of the microorganism, the microorganism promotes the oxidation / decomposition of the organochlorine compound in the soil. It can be understood that the organic chlorine compounds in the soil are efficiently decomposed.

Example 10 Acrylic acid crosslinked lipophilic polymer fine powder was charged with a ceramic porous body whose surface was heated, and the surface temperature was about 180 to 200 ° C. and the acrylic acid crosslinked lipophilic polymer. The fine powder of 1. was brought into contact with the powder to obtain a carrier in which the lipophilic polymer was thermally fused to the ceramic porous body.

On the other hand, YMCT-001 strain was cultured in LB medium to give OD 6
After 60 reached about 1.0, 100 g of the carrier was added to this solution with gentle stirring, and this was depressurized in a pressure bottle to introduce bacteria into the micropores of the carrier. The particle size of the lipophilic polymer was several hundred μm.

Next, this carrier was added to and mixed with 1000 g of unsterilized brown forest soil stored in a beaker having an internal volume of 500 ml, and then isolated from the soil and cultured in a Chalkley's salt medium to infect bacteria. capped with the protozoa (Colpoda sp.) suspension 40 ml (protozoa number 2 × 10 ⁴ cells) was added to filter paper. Then, 10 beakers treated in the same manner were prepared and allowed to stand in a constant temperature culture bath at 25 ° C.

Next, the number of YMCT-001 strains in the soil was measured for 10 days after the start of culture. The method for measuring the number of bacteria is
It was determined by the plate dilution method using a selective medium of YMCT-001 strain. Results are shown in FIG. In addition, YM in FIG.
The number of bacteria of CT-001 strain is an average value of 10 beakers.

(Example 11) 50 g of unsterilized brown forest soil whose TCE concentration was adjusted to 200 ppm was placed in a vial having an internal volume of 130 ml, and prepared in the same manner as in Example 10 to prepare a carrier holding bacteria. It was added to a vial and mixed with unsterilized brown forest soil. Then, the vial bottle was capped with a Teflon-coated butyl rubber cap, and further sealed with an aluminum cap. And the vial treated in the same way
20 pieces were prepared and left still in a constant temperature culture bath at 25 ° C.

Next, the TCE concentration in the gas phase in the vial after the start of culture was measured by the gas chromatograph headspace method. The number of YMCT-001 strains in the soil was measured by the same method as in Example 10. The results are shown in FIGS. 6 and 7.

(Embodiment 12) In the same manner as in Embodiment 10,
A carrier having a lipophilic polymer retaining YMCT-001 strain was obtained. This carrier is then added to a solution containing 100 ppm phenol.
The suspension was suspended in 1 M phosphate buffer, and the carrier lipophilic polymer absorbed phenol.

Then, the test was carried out under the same conditions as in Example 11 except that the lipophilic polymer as the carrier absorbed the phenol. The results are shown in FIGS. 6 and 7.

Example 13 TCE concentration in soil was 20 p
The test was conducted under the same conditions as in Example 11 except that the time was set to pm.

Table 4 shows the residual concentration of tetrachloroethylene 10 days after the start of the test.

Example 14 A test was conducted under exactly the same conditions as in Example 13 except that cis-dichloroethylene was added instead of tetrachloroethylene.

Table 4 shows the residual concentration of cis-dichloroethylene 10 days after the start of the test.

Example 15 Except that trans-dichloroethylene was added instead of tetrachloroethylene,
The test was performed under the same conditions as in Example 13.

Table 4 shows the residual concentration of trans-dichloroethylene after 10 days from the start of the test.

Example 16 A test was carried out under the same conditions as in Example 13 except that 1,1-dichloroethylene was added instead of tetrachloroethylene.

Table 4 shows the residual concentration of 1,1-dichloroethylene 10 days after the start of the test.

Example 17 Example 1 was repeated except that monochloroethylene was added instead of tetrachloroethylene.
The test was conducted under exactly the same conditions as in No. 3.

Table 4 shows the residual concentration of monochloroethylene after 10 days from the start of the test.

Example 18 PCB (Kanegafuchi Chemical Co., Ltd .: Kanechlor KC-30) was used instead of tetrachloroethylene.
The test was conducted under the same conditions as in Example 13 except that (0) was added.

Table 4 shows the residual concentration of PCB 10 days after the start of the test.

Comparative Example 10 Example 10 was repeated except that only the bacterial cells were separated from the suspension of the YMCT-001 strain by centrifugation and the bacterial cells were suspended in 0.1 M phosphate buffer and added to the soil.
The test was conducted under exactly the same conditions as above. Results are shown in FIG.

Comparative Example 11 Example 11 was repeated except that only the bacterial cells were separated from the suspension of the YMCT-001 strain by centrifugation and the bacterial cells were suspended in 0.1 M phosphate buffer and added to the soil.
The test was conducted under exactly the same conditions as above. The results are shown in FIGS. 6 and 7.

Comparative Example 12 Example 13 was repeated, except that only the bacterial cells were separated from the suspension of YMCT-001 strain by centrifugation and the bacterial cells were suspended in 0.1 M phosphate buffer and added to the soil.
The test was conducted under exactly the same conditions as above.

Table 4 shows the residual concentration of tetrachloroethylene 10 days after the start of the test.

Comparative Example 13 Example 14 was repeated except that only the bacterial cells were separated from the suspension of YMCT-001 strain by centrifugation and the bacterial cells were suspended in 0.1 M phosphate buffer and added to the soil.
The test was conducted under exactly the same conditions as above.

Table 4 shows the residual concentration of cis-dichloroethylene 10 days after the start of the test.

Comparative Example 14 Example 15 was repeated, except that only the bacterial cells were separated from the suspension of YMCT-001 strain by centrifugation and the bacterial cells were suspended in 0.1 M phosphate buffer and added to the soil.
The test was conducted under exactly the same conditions as above.

Table 4 shows the residual concentration of trans-dichloroethylene 10 days after the start of the test.

Comparative Example 15 Example 16 was repeated except that only the bacterial cells were separated from the suspension of YMCT-001 strain by centrifugation and the bacterial cells were suspended in 0.1 M phosphate buffer and added to the soil.
The test was conducted under exactly the same conditions as above.

Table 4 shows the residual concentration of 1,1-dichloroethylene 10 days after the start of the test.

Comparative Example 16 Example 17 was repeated except that only the bacterial cells were separated from the suspension of the YMCT-001 strain by centrifugation and the bacterial cells were suspended in 0.1 M phosphate buffer and added to the soil.
The test was conducted under exactly the same conditions as above.

Table 4 shows the residual concentration of monochloroethylene after 10 days from the start of the test. Comparative Example 17 Exactly the same as Example 18 except that only the bacterial cells were separated from the suspension of YMCT-001 strain by centrifugation and the bacterial cells were suspended in 0.1 M phosphate buffer and added to the soil. The test was conducted under the conditions.

Table 4 shows the residual concentration of PCB 10 days after the start of the test.

[0164]

[Table 4] As is clear from Example 10 and Comparative Example 10, microorganisms are held in a carrier provided with a lipophilic polymer, and the carrier is charged into the soil to bring the microorganisms into contact with the organic compound, thereby charging the carrier into the soil. It can be understood that it is possible to prevent predation of the microorganisms thus produced from the protozoa and to sufficiently maintain the survival of the microorganisms in the soil.

Further, Examples 11 and 12 and Comparative Example 11
As is clear from the above, when microorganisms are retained on a carrier equipped with a lipophilic polymer, the lipophilic polymer absorbs organic compounds even if a high concentration of organic compounds that is unsuitable for the survival of microorganisms is present in the soil. It can be seen that the microorganisms can be protected from the effects of organic compounds for retention and the survival of the microorganisms in the soil can be sufficiently maintained. Furthermore, by impregnating the lipophilic polymer provided in the carrier with the inducer, the oxidation and decomposition of the organic chlorine compounds in the soil by microorganisms are promoted, and the organic chlorine compounds in the soil can be efficiently decomposed. It can be understood.

Furthermore, Examples 13 to 18 and Comparative Example 1
As is clear from 2 to 17, by holding a microorganism in a carrier provided with a lipophilic polymer, and introducing this carrier into soil to bring the microorganism into contact with an organic compound,
It can be understood that many organic chlorine compounds other than are efficiently decomposed.

Example 19 YMCT-001 strain was cultured in LB medium, and after reaching OD 660 of about 1.0, 300 mg of a carrier made of liquefied wood porous foam was added to 200 ml of this solution. This was decompressed in a pressure bottle to introduce the bacterial cells into the fine pores of the carrier.

Next, this carrier was put into 25 ml of 0.1 M phosphate buffer in a vial, and trichlorethylene (T
CE) was added to 20 ppm and the vial was sealed with butyl rubber septum.

Next, the YMCT-001 strain was shake-cultured by shaking the vial at 25 ° C. and 100 rpm, and the TCE concentration in the vial was measured with time. The TCE concentration was measured by the headspace method by measuring the TCE concentration in the gas phase with a PID (photoionization detector) using a gas chromatograph. The results are shown in FIG.

Example 20 A test was conducted under exactly the same conditions as in Example 19 except that cis-dichloroethylene was added in place of trichloroethylene so that the concentration was 1 ppm.

Table 5 shows the residual concentration of cis-dichloroethylene after 4 days from the start of the test.

Example 21 A test was conducted under the same conditions as in Example 20, except that trans-dichloroethylene was added instead of cis-dichloroethylene.

Table 5 shows the residual concentration of trans-dichloroethylene after 4 days from the start of the test.

Example 22 Except that 1,1-dichloroethylene was added instead of cis-dichloroethylene,
The test was performed under the same conditions as in Example 20.

Table 5 shows the residual concentration of 1,1-dichloroethylene after 4 days from the start of the test.

Example 23 A test was conducted under the same conditions as in Example 20, except that monochloroethylene was added instead of cis-dichloroethylene.

Table 5 shows the residual monochloroethylene concentration after 4 days from the start of the test. (Example 24) A test was conducted under exactly the same conditions as in Example 20, except that PCB (Kanegafuchi Chemical Co., Ltd .: Kanechlor KC-300) was added instead of cis-dichloroethylene.

In Table 5, P after 4 days from the start of the test
The remaining concentration of CB is shown.

Example 25 A test was conducted under exactly the same conditions as in Example 20, except that tetrachloroethylene was added instead of cis-dichloroethylene.

Table 5 shows the residual concentration of tetrachloroethylene after 4 days from the start of the test.

(Example 26) Foam of liquid wood and diameter 1
A porous ceramic having a size of about mm was mixed and these were heat-fused to obtain a carrier in which a porous foam of liquid wood and the porous ceramic were composited.

A test was conducted under the same conditions as in Example 19 except that this carrier was used instead of the liquid wood porous foam. The results are shown in Fig. 8.

(Example 27) After the soil was put into the entire container having an internal volume of about 0.4 m ³ , the TCE concentration in the soil was 1 ppm on average.
TCE was added to the extent. Next, Example 19
The same carrier as that prepared in 1., which holds the YMCT-001 strain, was added to the soil. Then, while maintaining this container at 25 ° C., the TCE concentration in the container was measured with time in the same manner as in Example 19.

The results are shown in FIG.

(Comparative Example 18) A test was conducted under the same conditions as in Example 19 except that a porous foam of liquefied wood that did not retain the YMCT-001 strain was used.

The results are shown in FIG.

(Comparative Example 19) A test was conducted under exactly the same conditions as in Example 20 except that a porous ceramics carrier was used.

Table 5 shows the residual concentration of cis-dichloroethylene after 4 days from the start of the test.

(Comparative Example 20) A test was conducted under exactly the same conditions as in Example 21 except that trans-dichloroethylene was added instead of cis-dichloroethylene.

Table 5 shows the residual concentration of trans-dichloroethylene after 4 days from the start of the test.

COMPARATIVE EXAMPLE 21 Except that 1,1-dichloroethylene was added instead of cis-dichloroethylene.
The test was conducted under the same conditions as in Example 22.

Table 5 shows the residual concentration of 1,1-dichloroethylene after 4 days from the start of the test.

(Comparative Example 22) A test was conducted under the same conditions as in Example 23 except that monochloroethylene was added instead of cis-dichloroethylene.

Table 5 shows the residual monochloroethylene concentration after 4 days from the start of the test. (Comparative Example 23) A test was carried out under the same conditions as in Example 24 except that PCB (Kanegafuchi Chemical Co., Ltd .: Kanechlor KC-300) was added instead of cis-dichloroethylene.

In Table 5, P after 4 days from the start of the test
The remaining concentration of CB is shown.

(Comparative Example 24) A test was conducted under the same conditions as in Example 25 except that tetrachloroethylene was added instead of cis-dichloroethylene.

Table 5 shows the residual concentration of tetrachloroethylene after 4 days from the start of the test.

[0198]

[Table 5] (Comparative Example 25) A test was conducted under exactly the same conditions as in Example 19 except that a composite of a porous foam of liquefied wood and a porous ceramic, which did not hold the YMCT-001 strain, was used.

The results are shown in FIG.

(Comparative Example 26) A test was conducted under the same conditions as in Example 19 except that a porous ceramics carrier was used.

The results are shown in FIG.

(Comparative Example 27) A test was carried out under the same conditions as in Example 27 except that a porous foam of liquefied wood not holding the YMCT-001 strain was used.

The results are shown in FIG.

Examples 19, 26 and Comparative Examples 18, 25, 2
6 and Examples 20 to 25 and Comparative Examples 19 to 24, by holding the microorganisms in the porous foam formed by foaming liquefied wood and bringing the organochlorine compound into contact with the microorganisms, It can be understood that the organochlorine compound can be efficiently decomposed without re-administering the additive such as the aromatic compound to the microorganism.

Further, as is apparent from Example 27 and Comparative Example 27, by retaining the microorganisms in the porous foam formed by foaming liquefied wood and bringing the organochlorine compound into contact with the microorganisms, It can be understood that many organochlorine compounds other than TCE in soil are efficiently decomposed without re-administering additives such as aromatic compounds to microorganisms.

In the examples, an organic chlorine compound was used as the organic compound, but it goes without saying that many other compounds can be applied as the organic compound.

[0207]

As described above in detail, according to the method for activating a microorganism in the first invention of the present application, the first metabolic system for carrying out the assimilation-type reduction reaction and the second metabolism for carrying out the oxidation reaction. A substance serving as a substrate for the assimilation-type reduction reaction is administered to a microorganism having a system, and the first metabolic system is activated, and the activated first metabolic system causes the second metabolic system to Since it is activated, it can be applied not only in closed systems but also in open systems, and can activate microorganisms without administering chemical substances such as inducers that induce oxidase to organic compounds, giving to the environment. It is possible to provide a method for activating a microorganism with almost no influence.

Further, according to the method for decomposing an organic compound in the second aspect of the present invention, a microorganism having a first metabolic system for carrying out an assimilation-type reduction reaction and a second metabolic system for carrying out an oxidation reaction is provided. , A microorganism and an organic compound in which a substance serving as a substrate for the assimilation-type reduction reaction is administered to activate the first metabolic system, and the second metabolic system is activated by the activated first metabolic system Since it is contacted with, the microorganisms are activated not only in the closed system but also in the open system without administering an inducer or the like that induces an oxidase for the organic compound, and the organic compound such as the organic chlorine compound is efficiently oxidized / oxidized. It is possible to provide a method of decomposing an organic compound that can be decomposed and that has almost no influence on the environment.

Furthermore, according to the method for decomposing an organic compound in the third invention of the present application, a microorganism having a metabolic system for carrying out an oxidation reaction is carried on a carrier having a lipophilic polymer, and the carrier carrying the microorganism is carried out. Since the organic compound and the organic compound are contacted with each other, the microorganism is maintained at a high density and the organic compound is concentrated around the microorganism, and when the microorganism is introduced into the environment, it is possible to prevent the reduction of the microorganism and to efficiently It is possible to provide a method for decomposing an organic compound capable of oxidizing and decomposing an organic compound such as an organic chlorine compound. Further, according to the method for decomposing an organic compound in the fourth invention of the present application, a microorganism having a metabolic system that executes an oxidation reaction,
The carrier is carried on a carrier having a porous foam formed by foaming of liquefied wood, and the carrier carrying the microorganism is brought into contact with the organic compound, so that the microorganism is retained at a high density and is assimilated from the carrier to the microorganism. As substances elute, it is possible to activate microorganisms without administering an inducer that induces oxidase to organic compounds, and to oxidize and decompose organic compounds such as organochlorine compounds efficiently, which gives to the environment. It is possible to provide a method for decomposing an organic compound with almost no influence.

Further, according to the carrier of the fifth aspect of the present invention, since the liquefied wood is foamed and is provided with the porous porous foam having the constituent components of the wood, the microorganisms retained at a high density are It is possible to provide a carrier which is activated from the above and prevents the reduction of microorganisms when introduced into the environment, and which is a place for efficient oxidation and decomposition of organic compounds such as organic chlorine compounds. .

Further, according to the decomposition system for an organic compound in the sixth invention of the present application, a microorganism comprising an assimilation microorganism having a first metabolic system for carrying out an assimilation-type reduction reaction and a second metabolic system for carrying out an oxidation reaction. Since the substance that serves as a substrate for the type-type reduction reaction and the organic compound are contacted with each other, the microorganism is activated and the organic compound such as an organic chlorine compound is oxidized without administering an inducer that induces an oxidase of the organic compound to the microorganism. It is possible to provide a decomposition system of an organic compound which can be decomposed and can be applied not only to a closed system but also to an open system.

Therefore, the first to sixth inventions of the present application are excellent in economic efficiency and have extremely great industrial value.

[Brief description of drawings]

FIG. 1 is a schematic diagram showing an example of a decomposition system for an organic compound according to the present invention.

FIG. 2 is a schematic diagram showing an embodiment of a decomposition system for an organic compound according to the present invention.

FIG. 3 is a graph showing changes in TCE concentration with time in Examples 1, 8 and Comparative Example 1.

FIG. 4 is a graph showing changes in TCE concentration over time in Example 9, Comparative Example 8 and Comparative Example 9.

FIG. 5 is a diagram showing changes in the number of bacteria over time in Example 10 and Comparative Example 10.

FIG. 6 is a graph showing changes in the number of bacteria over time in Examples 11, 12 and Comparative Example 11.

FIG. 7 is a graph showing changes in TCE concentration with time in Examples 11, 12 and Comparative Example 11.

FIG. 8 is a graph showing changes in TCE concentration over time in Example 19, Example 26, Comparative Example 18, Comparative Example 25, and Comparative Example 26.

9 is a graph showing changes in Example 27 and Comparative Example 27 with time.
The figure which shows the change of TCE concentration.

[Explanation of symbols]

1 ... Pumping pump 2 ... Groundwater layer 3 ... Pumping well 4 ... Addition device 5 ... Introduction device 6 ... Injection well 7
… Coupling pipe 8… Groundwater level 9… Impermeable layer 10… Bioreactor 11… Adding device 12… Introduction pipe 13 ……… Recirculation pipe

─────────────────────────────────────────────────── ─── Continued Front Page (51) Int.Cl. ⁷ Identification Code FI B09C 1/10 C12S 13/00 C02F 11/02 ZAB C12N 1/20 C12S 13/00 C12R 1:01 // (C12N 1/20 B09B 3/00 E C12R 1:01) ZABA (72) Inventor Masaaki Morita 1 Komukai Toshibacho, Komukai-ku, Kawasaki-shi, Kanagawa Toshiba Research Consulting Co., Ltd. (56) Reference JP-A-6-226230 (JP, JP, 226230) A) JP-A-6-212155 (JP, A) JP-A-1-265881 (JP, A) JP-A-6-335926 (JP, A) (58) Fields investigated (Int. Cl. ⁷ , DB name) ) C12N 1/20 C02F 11/02 B09C 1/10 B09B 3/00

Claims (2)

(57) [Claims] 1. A YMCT-001 strain, which is a microorganism having a metabolic system that executes an oxidation reaction (Institute of Biotechnology, Institute of Biotechnology, Institute of Industrial Science and Technology).
Laboratory deposit FERM BP-5282 ) is supported on a carrier containing a porous body having a lipophilic polymer, the carrier carrying the microorganism is brought into contact with an organic compound, and the microorganism having the metabolic system is used. A method for decomposing an organic compound, which comprises decomposing the organic compound. 2. A YMCT-001 strain, which is a microorganism having a metabolic system that executes an oxidation reaction (Institute of Industrial Science and Technology
Laboratory deposit FERM BP-5282 ) is supported on a carrier having a porous foam formed by foaming of liquefied wood, and the carrier carrying the microorganism is brought into contact with an organic compound to provide the metabolic system. A method for decomposing an organic compound, characterized in that the organic compound is decomposed by the microorganism described above.