JP2000167532A - Decomposition of harmful environmental pollutant using edible mushroom - Google Patents

Abstract

PROBLEM TO BE SOLVED: To decompose pollutants without affecting the human body and an ecological system by charging edible mushroom hyphae, cultured hyphae and/or a mushroom bed into polluted soil or a water area. SOLUTION: In the decomposition of PCB and/or dioxins as organic chlorine compounds, edible mushroom hyphae, cultured hyphae and/or a mushroom bed are charged into polluted soil or a water area. In this process, lignosulfonic acid or the sulfonate substance of a lignin-containing substance is charged during the charging or before/after the charging. The edible mushroom to be used is selected from gifola fronlosa, dentinula edodes, dyephyllum shimeji, and Flammulina volutipers, and at least one kind of Maitake selected from white-Maitake, kite-Maitake, and others is used preferably. The lignosulfonic acid or the sulfonated substance of a lignin-containing substance can be charged into the soil or the water area simultaneously with the hyphae and occasionally after the charging into the hyphae and others.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a method for decomposing harmful environmental pollutants using a mycelium of an edible mushroom, a mycelium culture, a fungal bed and / or a waste bacterial bed after cultivation.

[0002]

2. Description of the Related Art Recent environmental investigations have revealed extensive contamination of harmful and hardly decomposable aromatic compounds, and there is a concern that these environmental pollutants may have an adverse effect on ecosystems. Since these hard-to-degrade chemicals are not easily degraded by microorganisms, they have accumulated in polluted soil and water for a long period of time, and the development of harmful environmental pollutant decomposition treatment technology to restore the polluted environment has been developed. It is strongly desired.

As a technique for restoring a polluted environment, a technique has been developed to decompose and detoxify pollutants by increasing the ability of microorganisms in the soil to decompose, and this is achieved by enhancing the self-cleaning capacity of ecosystems. The aim is to promote the decomposition of. However, when the concentration of the pollutant in the soil is high, or when the pollutant is an extremely hardly decomposable substance, the self-cleaning action of the ecosystem alone cannot often provide a sufficient effect. If sufficient purification is not recognized only by the self-cleaning action of the ecosystem, a method of actively administering useful microorganisms having the ability to decompose pollutants to the contaminated site may be considered.

[0004] In a closed system such as a sewage treatment plant, microorganisms such as activated sludge are actively used in practice, and the effect is given. However, the release of degrading microorganisms into open systems has not been accepted due to the problem of public acceptance. Recently, a high concentration of pollution in a wide range has been elucidated by harmful chemical substances such as trichlorethylene and tetrachlorethylene in the underground waters, such as a semiconductor plant, the activity of the self-cleaning action of indigenous microorganisms by supply of nutrients in these cases The effect is not recognized enough.

[0005] Polychlorinated biphenyls (PCs), which are extremely difficult to decompose among organochlorine compounds, especially chlorinated aromatic compounds,
In the case of contamination by B), as with dioxins such as polychlorinated dibenzo-p-dioxin (PCDD) and polychlorinated dibenzofuran (PCDF), they are not easily susceptible to degradation by natural microorganisms. Use is necessary. So far, the genus Pseudomonas , Alcaligenes
Bacteria that degrade PCBs, such as the genus Acinetobacter, have been isolated from nature, but most of these PCB-degrading bacteria have a large number of chlorine substitutions (for example, 4 or more chlorine substitutions).
The decomposition ability was low, and only the PCB with a limited structure was decomposed.

[0006] In recent years, it has been confirmed that some Rhodococcus bacteria degrade PCB isomers up to high chlorine substitution. However, addition of a high concentration of PCB (eg, 100 ppm) reduces cell growth and PCB resolution. It was also clarified that it was completely inhibited, and problems in practical use of environmental restoration by microorganisms have been pointed out (M. Seto et al. Biotechnology Let
ters, Vol. 18, No. 11, p1305-1308 (1996)).

Furthermore, a particular problem in the degradation of PCBs or dioxins by microorganisms is the mixture of various substituted chlorine numbers and substituted chlorine positions (theoretically, there are 209 types of PCBs, PCDD and PCDF of dioxins). In addition, treatment of PCBs or dioxins using a degrading enzyme with a limited substrate specificity of degrading microorganisms, which have already been confirmed, is unsuitable for application to environmental purification because many residual components are not treated. Was enough.

[0008]

SUMMARY OF THE INVENTION An object of the present invention is to provide a method capable of achieving the removal of harmful environmental pollutants in soil or water bodies without adversely affecting the human body and ecosystem.

[0009]

[Means for Solving the Agenda] The inventors of the present invention have conducted intensive studies to solve the above-mentioned problems, and as a result, the mycelium, mycelium culture, and fungal bed of edible mushrooms, especially the waste bacterial bed after cultivation, are effective. They found something and completed the present invention. That is, the present invention relates to (1) edible mushroom mycelium, mycelium culture or /
And a method for decomposing a harmful environmental pollutant, which comprises decomposing a contaminant by administering the fungal bed to contaminated soil or water,

(2) The edible mushrooms are maitake, shiitake,
(1) The method for decomposing harmful environmental pollutants according to (1), which is any of Bunashimeji and Enokitake.
(3) Maitake ( Grifola frondosa ), White Maitake ( Grifola albicans ), Choreimaitake ( Grif
ola umbellatus ), Tobimaitake ( Grifola gigante)
a ) The method for decomposing harmful environmental pollutants described in (2), which is at least one of the following: (4) The harmful environment described in (1), wherein the pollutant is an organic chlorine compound. Pollutant decomposition treatment method,

(5) The method for decomposing harmful environmental pollutants according to (4), wherein the organochlorine compound is PCB and / or dioxins, (6) a mycelium, a mycelium culture or / and / or an edible mushroom A method for decomposing harmful environmental pollutants, which comprises adding or administering a lignosulfonic acid or a lignin-containing substance sulfonate at or before or after administration, upon administering the bacterial bed to the contaminated soil or water area, (7) A method for decomposing harmful environmental pollutants, which comprises decomposing contaminants by administering a waste bacterial bed after cultivation of edible mushrooms to contaminated soil or water.

(8) When the waste fungus bed after cultivation of the edible mushroom is administered to the contaminated soil or water area, lignosulfonic acid or a sulfonate of a lignin-containing substance is added or administered before or after the administration. (7) The method for decomposing harmful environmental pollutants according to (7), (9) The waste bacteria bed after cultivation of edible mushrooms is cut into blocks of an appropriate size or cut into small pieces or powder, The present invention relates to the method for decomposing and processing harmful environmental pollutants according to (7) or (8), which is laid, buried or sprayed on contaminated soil.

The present invention has been made based on the following findings. In other words, it is a kind of lignin-degrading basidiomycete
Phanerochaete chrysosporium was found to degrade harmful chlorinated aromatic chemicals such as dioxin, and it was reported that there was a correlation between the peroxidase activity secreted by this strain and the degradation of dioxin. Yes (Tachibana et al., Journal of Japan Pulp and Paper Technology, Vol. 50, No. 12, P180
6-1815, (1996)).

On the other hand, cultivation of mushrooms requires addition of sawdust to the culture medium, but the mycelium of mushrooms decomposes hard-to-degrade lignin that gives strength to wood using enzymes such as peroxidase and laccase. It has been revealed that. These enzymes, together with H ₂ O ₂ , are thought to degrade the three-dimensional lignin network in a non-specific manner, but recently Phanerochaete chryso
These enzymes secreted by sporium have been shown to degrade a wide range of harmful chlorine chemicals such as dioxin (Takada et al. Appl. Environ. Microbiol., vo
l. 62, p4323 (1996)). However, it is not always sufficient in terms of safety and quantity.

The present inventors have thought that edible mushrooms may also have the ability to decompose harmful chemical substances, and carried out as described below. As a result, satisfactory results were obtained. In order to enable bioremediation in an actual contaminated environment (open system), it is an important factor not only that the strain has excellent resolution but also that its safety as a strain has been confirmed. However, also in this respect, the safety of the mushroom hypha or the waste bacterial bed after mushroom cultivation has been confirmed, so that the problem of public acceptance in use in an open system is well tolerated.

[0016]

DETAILED DESCRIPTION OF THE INVENTION In the present invention, edible mushroom mycelium, mycelium culture, and fungal bed are used. The fungus bed includes everything from planting mushroom mycelium for mushroom cultivation to waste bacterial bed after mushroom cultivation. Environmental pollution is widespread, and it is ideal to decompose these pollutants using inexpensive and mass-produced products. Mushrooms are produced in large quantities in Japan, It is economical to use the waste bacteria bed and it is two birds with one stone.

In the present invention, edible mushrooms such as maitake, shiitake, bunashimeji and enokitake are recently produced in bag cultivation and bottle cultivation using sawdust, and for example, in the case of bag cultivation, After harvesting the fruiting body of the mushroom, the bag is removed from the block-shaped waste fungus bed and then cut as it is or cut into blocks of an appropriate size and laid or buried in the contaminated soil. In the case of laying, it may be simply placed on the soil, but it is preferable to dig and fix a hole such that the block slightly comes out or becomes slightly above the ground surface. Further, the soil may be sprinkled thereon.

If the contaminated soil covers a wide area, it is effective to bury a block of the waste bacteria bed deeper so as to surround the contaminated area. Also, depending on the situation of contamination, two layers, three
It can also be embedded in layers and multilayers. The waste bacteria bed can be scattered and buried in contaminated soil as small pieces or powder in addition to the block form. The waste bacterial bed can maintain favorable conditions under which the mushroom hypha can grow in the soil, and is easily compatible with the soil, and is suitable for a method of treating contaminated soil, which is called “individual treatment” or “in situ treatment”.

On the other hand, the fungal hyphae of the mushrooms
Alternatively, a culture of mushroom mycelium can be administered to a contaminated water area or an environment such as soil or collected contaminated soil to decompose harmful pollutants. For example, add appropriate water to the contaminated soil, make it into a slurry, and transfer it to the reaction tank.
Add mushroom mycelium and nutrients or mushroom mycelium culture,
The harmful contaminants can be decomposed ("slurry processing") by stirring.

It is considered effective to add in advance a substance which induces an enzyme for decomposing harmful pollutants to the mycelium of the edible mushroom, the mycelium culture, the fungal bed, and the waste bacterial bed after cultivation. As a result of various studies, the present inventors have found that sulfonic oxides of lignosulfonic acid or lignin-containing substances (such as paper pulp) are useful. Further, in that case, it is more effective to add together with glucose as a carbon source and ammonium tartrate as a nitrogen source.

The lignosulfonic acid or the sulphonate of the lignin-containing substance may be administered to the soil or water at the same time as the mycelium or the like, or, in some cases, after administration to the mycelium or the like. In that case, glucose as a carbon source,
It may be administered together with a nutrient source for the growth of mushrooms such as ammonium tartrate serving as a nitrogen source. The addition amount may be about 0.1% based on the amount of the medium, the bacterial bed and the waste bacterial bed.

As edible mushrooms, maitake, shiitake, bunashimeji, enokitake, and the like can be used as described above. In particular, maitake is superior in decomposition processing ability to other mushrooms, and maitake is produced in a factory. As a result, a large amount of waste bacteria bed is produced, so that the quantity can be ensured, which is particularly preferable. Maitake ( Grifola fron)
dosa ), White Maitake ( Grifola albicans ), Choreimaitake ( Grifola umbellatus ), Tonmaimaitake ( Grif
ola gigantea ) and maitake ( Grifola)
frondosa ) and white maitake ( Grifola albicans ) are particularly preferred.

The harmful environmental pollutants according to the present invention include, for example, chlorinated aromatic compounds such as dioxins such as PCB, PCDD and PCDF, organic chlorinated pesticides such as DDT and its analogous substances hexachlorocyclohexane and dieldrin, trichloroethylene, tetrachloroethylene, and the like. Chlorine-substituted organic solvents such as carbon tetrachloride and methyl chloride are exemplified. In addition, aromatic compounds such as benzene and toluene, and polycyclic aromatic hydrocarbons such as phenol, cresols, and creosote can be mentioned, and also substances that are generally considered to pollute the environment. Hereinafter, embodiments will be described, and embodiments and effects of the present invention will be described in more detail.

(Example 1) A waste bacterial bed after harvesting the fruit body of oyster mushroom was prepared, and 25 g of mycelial mass was aseptically added to 50 ml of an inorganic medium. 10,0 by Waring blender
After mixing the mycelial mass with ice cooling at 00 rpm for 10 minutes, the above mycelial suspension was added to a 15 ml sterilized culture test tube.
Each 5 ml was dispensed. 0.1 pp final concentration for mycelial suspension
Each of various PCB standard solutions was added by a microsyringe so as to obtain m, followed by shaking culture at 25 ° C. After culturing for 7 days,
The mycelial culture was mixed with a polytron-type handy mixer, and acidified by adding hydrochloric acid, and then the PCB components remaining in the culture were extracted with ethyl acetate. After concentration of extraction solvent to 100μl, with mass spectrometer
-The remaining PCB components were analyzed by gas chromatography. By comparing the amount of PCB remaining after culturing for 7 days using the sterilized mycelium suspension with the control system, the amount of PCB degradation by the maitake mycelium derived from the waste bacterial bed was calculated. Table 1 shows the results.

Composition of inorganic medium (in 1 liter); KH ₂ PO ₄ =
20g, NaH ₂ PO ₄ = 0.2g, MgSO ₄・ 7H ₂ O = 0.5g, CaCl ₂ = 100
_{μg, FeSO 4 · 7H 2 O} = 100μg, ZnSO 4 · 7H 2 O = 10μg, CuSO 4
・ 5H ₂ O = 20μg2,2-dimethyl succinate = 1.46g, Ammoniu
m tartrate = 0.2g, Thiamin hydrochloride = 100μg,

[0026]

[Table 1]

From the results, it was found that the maitake waste bacteria bed
It can be seen that the decomposition of PCB is similarly effectively decomposed even if the number of chlorine substitution is large. (Example 2) Maitake mycelium ( Grif
ola frondosa ) was cultured with shaking at 25 ° C for 2 weeks in 100 ml of potato dextrose medium (Difco). Hyphal colonies aseptically 10,000 rpm with Waring blender
For 10 minutes with ice cooling. After washing the mycelium pieces after mixing twice with the inorganic medium described in Example 1, the mycelium was suspended to a concentration of 0.1 g / 10 ml. Glucose (20 g / l) was added as a carbon source, and lignosulfonic acid (1 g / l) was added for the purpose of inducing a group of decomposition enzymes. PCB so that the final concentration is 10 ppm for the mycelium solution
48 (a PCB mixture mainly composed of 4-chlorine substitution) was added by a microsyringe, followed by shaking culture at 25 ° C.

Glucose (20 g / l) was added every 10 days. After culturing for 60 days, the mycelial culture was mixed with a polytron-type handy mixer, acidified by adding hydrochloric acid, and the PCB components remaining in the culture were extracted with ethyl acetate. After the extraction solvent was concentrated to 100 μl, the remaining PCB components were analyzed by gas chromatography with a mass spectrometer. By comparing the amount of PCB remaining after culturing for 60 days with the sterilized hypha culture medium with the control system, the amount of PCB degradation by the Maitake mycelium was calculated. Table 2 shows the results.

[0029]

[Table 2]

As can be seen from these results, the maitake mycelium also effectively reduced the PCB. (Example 3) Maitake mycelium ( Grif
ola frondosa ) in a 100 ml potato dextrose medium (Difco) at 25 ° C. with shaking for 2 weeks. Hyphal colonies aseptically 10,000 rpm with Waring blender
Mix for 10 minutes while cooling on ice. After washing the mycelium twice with the above inorganic medium, the mycelium is suspended to a concentration of 0.1 g / 10 ml. Glucose (20
g / l). 10ppm final concentration for mycelial suspension
PCB48 (PCB mixed solution mainly composed of 4-chlorine substitution) is added by a microsyringe so as to obtain a culture solution at 25 ° C. with shaking. Metabolic intermediates produced in the culture solution were sampled over time and analyzed by gas chromatography with mass spectrometer. As a result, dichloro-methoxy-p
The accumulation of henol was clarified, and it was clarified that PCB was degraded to monocyclic compound by Maitake hyphae.

[0031]

According to the present invention, the mycelia of edible mushrooms, mycelial cultures, fungal beds and / or waste bacterial beds after cultivation are administered to soil or water bodies contaminated with harmful environmental pollutants. In addition, a useful group of degrading enzymes produced by edible mushroom mycelia has made it possible to safely and effectively decompose harmful environmental pollutants.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) C12N 1/14 C12N 1/14 E (72) Inventor Masao Fukuda 1769-1 Fukasawacho, Nagaoka City, Niigata Prefecture (72 ) Inventor Yasuo Ohira 2610-4 Yokawa, Minamiuonuma-gun, Niigata Pref. 4D004 AA41 AB06 AB07 AC07 CA17 CC15 4D040 DD00 DD11

Claims (9)

[Claims] Claims 1. An edible mushroom mycelium, a mycelium culture or
And / or administering the bacterial bed to the contaminated soil or water area to decompose the contaminants. 2. The method for decomposing harmful environmental pollutants according to claim 1, wherein the edible mushroom is one of Maitake, Shiitake, Bunashimeji and Enokitake. [3] The maitake is a maitake ( Grifola frondos)
a ), White Maitake ( Grifola albicans ), Choreimaitake ( Grifola umbellatus ), Tonmaimaitake ( Grifol )
a gigantea ), the method for decomposing harmful environmental pollutants according to claim 2, wherein 4. The method according to claim 1, wherein the pollutant is an organic chlorine compound. 5. The method for decomposing harmful environmental pollutants according to claim 4, wherein the organic chlorine compound is PCB and / or dioxins. 6. An edible mushroom mycelium, mycelium culture or
And / or when administering a bacterial bed to contaminated soil or water, adding or administering a lignosulfonic acid or a sulfonate of a lignin-containing substance at or before or after the administration, and decomposing harmful environmental pollutants. Method. 7. A method for decomposing a harmful environmental pollutant, which comprises decomposing a pollutant by administering a waste fungal bed after cultivation of edible mushrooms to contaminated soil or water. 8. The method of administering a waste fungus bed after cultivation of edible mushrooms to contaminated soil or water, adding or administering lignosulfonic acid or a sulfonate of a lignin-containing substance before or after administration. The method for decomposing harmful environmental pollutants according to claim 7, characterized in that: 9. The method according to claim 1, wherein the waste bacterial bed after cultivation of the edible mushrooms is cut into blocks of an appropriate size, or cut into small pieces or powder, and laid, buried or sprayed on the contaminated soil. The method for decomposing harmful environmental pollutants according to claim 7 or 8.