TWI594815B - Volatile organic halogen compound - Google Patents

Description

Decomposition accelerator for volatile organic halogen compounds

The present invention relates to a decomposition accelerator and a decomposition promoting method for a volatile organic halogen compound, and more particularly to a decomposition accelerator capable of promoting decomposition of a microorganism of a volatile organic halogen compound, and a decomposition promoting method using the same.

The soil or groundwater of the land is sometimes contaminated with natural or artificial chemicals. The pollution of such land does not need to be said about the use of the land as a farmland for food production. There are also major problems in the case of attempted residence or commercial use.

However, unlike direct oral foods, or clothing that touches the skin, cosmetics, decorations, etc., land pollution, especially soil pollution, is not of much concern. In the past, there was also a hole in the landfill chemical to deal with it. example.

The purification of such land pollution has recently become a major problem, and various purification treatment methods have been proposed for this.

If the method of purifying the land is classified as a large item, it can be classified into a physics that decomposes the chemical substance by high temperature and adsorbs it on activated carbon. Sexual treatment, chemical treatment to make chemical substances harmless by chemical reaction, microbial purification method using microorganisms capable of decomposing chemical substances, and plant purification method using plants having the ability to absorb or adsorb chemical substances.

Among them, the use of microorganisms or plants to suppress the cost or cost to the environment has recently attracted attention, and the purification method using microorganisms is also called "bioremediation", and the purification method using plants is also called "phytoremediation". Various research and development are carried out, and the research and development of bioremediation is active because the soil that can be processed is large.

There are 2 techniques for bioremediation. One is to use a technique in which a microorganism that has an effect on the decomposition of a target pollutant is preliminarily confirmed to be in a contaminated place, and is called biofortification. The other is a technique for activating the action of microorganisms by giving an indigenous microbial oxygen or nutrient source to a contaminated site, thereby promoting purification, called biostimulation.

Also, there are 2 methods of bioremediation. First, the method of removing contaminated soil or groundwater and treating it in other places is called "facility treatment." The other is that the method of purifying contaminated soil or groundwater at the site is called "original location purification."

In recent years, pollutants that have become a major problem include organic halogen compounds represented by organochlorine compounds such as tetrachloroethylene, trichloroethylene, dichloroethylene, dioxin, and polychlorinated biphenyls. Among them, volatile organic halogen compounds such as tetrachloroethylene, trichloroethylene, and dichloroethylene have the concern of affecting the human body through respiration, and it is common to have to deal with it quickly.

This volatile organic halogen compound is easily saturated into the soil, When reaching the groundwater vein, it is easy to expand the pollution to a wide range. Although the soil microorganisms decomposing such volatile organic halogen compounds also exist, generally the soil is closer to the surface, the more organic matter is present, the soil microorganisms are also abundant, and the organic matter and soil microorganisms are reduced from the surface layer to the deeper, if the depth is more than 1 m. Then the activity of the microorganism is reduced to less than 1/100 of the surface layer. Due to such conditions, once the land contaminated with the volatile organic halogen compound has a problem of being relatively low-concentration and extensively contaminated for a long time.

Since such a relatively low concentration and purification of a large-scale contaminated land is effective as a means of bioremediation, various proposals for purifying the contamination of volatile organic halogen compounds have been proposed.

For example, biostimulation using a composition containing an ester of polylactic acid and a polyfunctional alcohol such as glycerol, xylitol, sorbitol, or pentaerythritol (for example, refer to Patent Document 1); using yeast, fatty acid, and carbohydrate Biostimulation of a composition such as the above (see, for example, Patent Document 2); biostimulation using a product of a condensation reaction of an amino acid with an oxycarboxylic acid (see, for example, Patent Document 3) is proposed. Further, Non-Patent Document 1 discloses various methods for bringing a decomposition accelerator into contact with soil and/or groundwater in which microorganisms are present.

On the other hand, microorganisms used as a bacterium for purifying a soil contaminated with a volatile organic halogen compound are known as anaerobic bacteria, particularly bacteria of the genus Dehalococcoides. In the absence of this microorganism, the volatile organic halogen compound does not decompose to the final ethylene, but decomposes the dichloroethylene which may be stopped in the intermediate substance, and may not be completely purified. However, even bacteria such as dehalogens usually depend on their species. It is necessary to know that the decomposable compound is involved in the decomposition of tetrachloroethylene to ethylene with several kinds of dehalogen bacteria. (Refer to Non-Patent Document 2) Therefore, the decomposition rate of various volatile organic halogen compounds of the microorganisms of the genus Dehalogen is not necessarily fast, and even if the bacteria of the genus Dehalogen are present, they do not necessarily participate in the decomposition of the volatile organic halogen compounds.

Therefore, bio-enhancement using a mixed strain of a plurality of dehalogen bacteria (for example, refer to Non-Patent Document 3), or use of a dehalogen bacteria and a linked strain as a bio-enhancement of a main group (for example, a reference patent) Document 4) was proposed. However, the method of using this mixed strain also has a problem of delay in the purification rate depending on the soil contamination state or pH, and hence the organic matter content.

[Advanced technical literature] [Patent Literature]

[Patent Document 1] Japanese Patent Publication No. 2000-511969

[Patent Document 2] Japanese Patent Laid-Open Publication No. 2005-185870

[Patent Document 3] Japanese Patent Laid-Open Publication No. 2010-104962

[Patent Document 4] Japanese Laid-Open Patent Publication No. 2011-244769

[Non-patent literature]

[Non-Patent Document 1] Industrial Research Association of the Co., Ltd. "Chemical Devices" July 2007 issue, Yamazaki Yu "Soil ‧ Groundwater Purification Technology - VOC Decomposition and Purification Technology -"

[Non-Patent Document 2] Sakihara Sho, "In-situ of vinyl chloride as a target Analysis of the behavior of bacteria in the genus Dehalococcoides in bioremediation" Lectures on research on groundwater ‧ soil pollution and its prevention measures (2008)

[Non-Patent Document 3] Yagi Slim's "The Status Quo and Future Prospects of Land Remediation Technology" "Seeking Food and Environmental Safety - Harmful Chemical Substances in Agriculture, Forest and Water Production Systems - 10 Pages of Essentials" (2007)

However, the method described in Patent Document 1 or Patent Document 3 has low microbial activity at the initial stage of administration, and therefore has a low purification rate and is harmless for contaminated soil, particularly groundwater containing a large amount of volatile organic halogen compounds. The long time required for the process. Further, the method described in Patent Document 2 has an effect of promoting the decomposition reaction of the volatile organic halogen compound, but the activation of the microorganism is still insufficient, and the time required for the harmlessness is long. On the other hand, the method using the mixed strain disclosed in Non-Patent Document 3 or Patent Document 4 also has a problem that the purification rate is delayed depending on the soil contamination state or the pH and the organic matter content.

Accordingly, it is an object of the present invention to provide a decomposition accelerator which is a decomposition accelerator for purification (bioremediation) of microorganisms by a soil contaminated with a volatile organic halogen compound, in particular, by raising microorganisms in an initial stage of administration. The activity enables the volatile organic halogen compound to be quickly detoxified.

Still another object of the present invention is to provide a microorganism for promoting A method for decomposing a volatile organic halogen compound, which is a method for purifying a volatile organic halogen compound by performing microbial purification (bioremediation) of a land contaminated with a volatile organic halogen compound, in particular, by increasing microbial activity at an initial stage of administration Quickly harmless.

As a result of various investigations in order to achieve the above object, the inventors of the present invention have found that one or two or more of the following (A) to (C) can be effectively used as a decomposition accelerator for a volatile organic halogen compound. The present invention has been completed by promoting the decomposition of volatile organic halogen compounds by microorganisms.

In other words, the decomposition accelerator of the volatile organic halogen compound of the present invention is characterized by containing one or more of the following (A) to (C).

(A) the fruit of citrus or the extract obtained from the fruit

(B) citrus peel or extract obtained from the peel

(C) Formulations containing all of the following (1) to (3)

(1) Glycerin

(2) Milk protein and / or yeast extract

(3) Vitamin B12

Further, when the decomposition accelerator of the present invention contains at least one of (A) and (B), the extract is preferably an extract obtained by using water as a solvent.

When the decomposition accelerator of the present invention contains (C), it is contained in an amount of 0.1 to 3 parts by mass based on 1 part by mass of the glycerin of the above (1). (2) Milk protein and/or yeast extract is preferred.

Further, in the case where the decomposition accelerator of the present invention contains (C), it is preferable to contain 0.00001 to 0.001 parts by mass of the vitamin B12 of the above (3) with respect to 1 part by mass of the glycerin of the above (1).

The decomposition accelerator composition of the present invention is characterized by containing a decomposition accelerator as an active ingredient.

The method for promoting decomposition of a volatile organic halogen compound by microorganisms according to the present invention is characterized in that the decomposition accelerator or the decomposition accelerator composition of any of the above is brought into contact with soil and/or groundwater containing a volatile organic halogen compound.

In the method of the present invention, the volatile organic halogen compound is preferably an organic chlorine compound.

Further, in the method of the present invention, the organochlorine-based compound is preferably selected from the group consisting of carbon tetrachloride, chloroform, dichloromethane, methyl chloride, 1,2-dichloroethane, and vinylidene chloride. Cis-1,2-dichloroethylene, trans-1,2-dichloroethylene, 1,3-dichloropropene, tetrachloroethylene, 1,1,1-trichloroethane, 1,1,2-three At least one of a group of ethyl chloride, trichloroethylene, and vinyl chloride.

Further, in the method of the present invention, the microorganism is preferably selected from the group consisting of Clostridium bacteria, Dehalobacter bacteria, Dehalococcoides bacteria, and Dehalospirilum. At least one of a bacterium, a Desulfobacterium bacterium, a Desulfomonas bacterium, and a Desulfomonile bacterium.

The decomposition accelerator of the present invention is easy to handle, and the contaminated soil or groundwater can be quickly and harmlessly purified by using microorganisms for purification (bioremediation) of the soil contaminated with the volatile organic halogen compound, and the low-cost and environmental burden is not burdened. .

Further, the method of the present invention is low-cost and has no burden on the environment, and can promote purification of microorganisms in soil or groundwater contaminated with volatile organic halogen compounds.

Fig. 1 is a graph showing the transition of c-DCE, VC, and ethylene in Example 1.

Fig. 2 is a graph showing the transition of c-DCE, VC, and ethylene in Example 2.

Fig. 3 is a graph showing changes in c-DCE, VC, and ethylene amount in Comparative Example 1.

4 is a graph showing changes in c-DCE, VC, and ethylene amount in Comparative Example 2.

Fig. 5 is a graph showing changes in the amounts of trichloroethylene, c-DCE, VC, and ethylene in Example 7.

Fig. 6 is a graph showing changes in the amounts of trichloroethylene, c-DCE, VC, and ethylene in Comparative Example 3.

Fig. 7 is a graph showing the transition of the amount of trichloroethylene, dichloroethylene, vinyl chloride, and ethylene in Example 15.

Hereinafter, the decomposition accelerator of the volatile organic halogen compound of the present invention will be described in detail. Further, in the present specification, the decomposition of the volatile organic halogen compound means the dehalogenation of the volatile organic halogen compound.

The decomposition accelerator of the volatile organic halogen compound of the present invention is characterized by containing one or more of the following (A) to (C).

(A) the fruit of citrus or the extract obtained from the fruit

(B) citrus peel or extract obtained from the peel

(C) Formulations containing all of the following (1) to (3)

(1) Glycerin

(2) Milk protein and / or yeast extract

(3) Vitamin B12

First, the above components (A) and (B) will be described.

The type of the citrus of the above (A) component and (B) component is not particularly limited, and may be a plant of the genus Citrus subfamily of the genus Rutaceae, especially by the genus Citrus or the kumquat belonging to the genus Citrus. It is preferred to use a plant produced by crossing the citrus or kumquat or the like. Specific examples of the citrus can be, for example, night orange, navel orange, blood orange, grapefruit, lemon, pomelo, lime, citrus, gossip, licorice, mandarin, kumquat, orange, and the like. The oranges produced by the yummy, the clear, the fire, etc., or the orange pomelo, such as Seminole or Minneola. Among them, it is preferable to use one or more of the late orange, grapefruit, lemon, and mandarin orange in the point that it is easy to obtain and can be obtained in a large amount at a low price.

In the above-mentioned (A) component and (B) component, the fruit of the above-mentioned citrus may be one part of the fruit, but it is preferable that the peeling effect is high.

The form of the component (A) and the component (B) is not particularly limited, and may be, for example, a direct user of fruit, peel or pulp. The fruit, the peel or the pulp is dried, and the fruit, peel or pulp is pulverized and dispersed. Water, fruit, peel or pulp by powder, juice, etc. Further, an extract extracted from fruits, particularly a peel, with water such as warm water to hot water, or an extract obtained by using a polar solvent such as ethanol, acetone or ethyl acetate or a nonpolar solvent such as hexane may be used. .

Among them, it is preferable that the extract extracted from the peel from the warm water to the hot water or the like is effective in application to the soil and can obtain a high effect. Further, the temperature of the water at the time of extraction is preferably from 30 to 100 ° C, more preferably from 60 to 95 ° C, still more preferably from 60 to 80 ° C.

The above-mentioned extraction and separation apparatus for extraction may be a device which can efficiently obtain an extract which constitutes a decomposition accelerator of the volatile organic halogen compound of the present invention, and examples thereof include a continuous centrifugal device, a membrane separation device, and an ultra Critical extraction device, etc.

Moreover, the peel of citrus is a large amount of by-products when producing a processed product (juice, canned, etc.) of citrus fruits, and is mostly discarded without useful use in the past, and the decomposition accelerator of the volatile organic halogen compound of the present invention is used as The raw material does not only provide a product which is lower in cost than the decomposition accelerator of the volatile organic halogen compound known in the related art, and is meaningful from the viewpoint of efficient use of resources.

Further, the extract obtained in the form of one of the components (A) and (B) may be extracted from a residue obtained by extracting a component such as pectin, aroma components, a pigment, or hesperidin from a fruit, and further contains such a residue. Other ingredients are also available.

Further, since the citrus fruit is eaten since ancient times, the decomposition accelerator of the volatile organic halogen compound of the present invention which is used as a raw material has high safety and is relatively stable to heat and thus is easy to handle.

Among the citrus fruits, in particular, the peel of the citrus fruits exhibits a high decomposition-promoting effect, and the extract obtained by extracting water such as warm water or hot water has a high effect. Therefore, although the detailed reason is not clear, it is considered that one of the components contained in the citrus fruit which exerts the decomposition accelerator effect of the present invention is a mixture of water-soluble saccharides or salts and organic acids.

Next, the above component (C) will be described.

The component (C) is a formulation containing all of the following (1) to (3).

(1) Glycerin

(2) Milk protein and / or yeast extract

(3) Vitamin B12

The glycerin used in the preparation of the component (C) is a microbial carbon source and is a supply source of hydrogen for replacing the chlorine atom of the organochlorine-based compound, that is, a donor of hydrogen, and may be directly used as glycerin or in combination. The form of the glyceride of 1 to 3 fatty acids is preferably glycerol directly. When using commercially available glycerin, the purity is 100% or more (for example, the test drug is special), and the glycerin of the Japanese Pharmacopoeia can be used. 80 to 90%), or refined glycerin D, food additive glycerin, and concentrated glycerin for cosmetics (all manufactured by Lion Co., Ltd.).

In the formulation of the above component (C), milk protein and/or yeast extract is used as a nitrogen source of microorganisms.

As the milk protein described above, only whey protein, casein protein, casein protein, and whey protein may be used in combination, but whey protein and casein protein are preferably used in combination.

Further, the milk protein is preferably water-soluble. When a commercially available product is used, it may be a product containing a milk protein at a high concentration, and it may be harmless to the human body (or a person who does not significantly hinder the growth of the microorganism), for example, sodium caseinate or casein. Potassium, whey powder, WPC (whey protein concentrate), WPI (whey protein isolate), whole milk protein (TMP), protein concentrate whey powder, whole milk powder, skim milk powder, lactose whey, Lactose-free whey powder, cheese powder, sugared milk powder, prepared milk powder, milk protein concentrate (MPC), and the like. In the present invention, in view of low lipid content and high preservation stability, whole milk protein (TMP) and/or skim milk powder are preferred, and skim milk powder is more preferable.

The yeast extract refers to an extract extracted by self-digesting a yeast culture or by an enzyme, hot water, physical crushing, acid decomposition, alkali decomposition, freeze-thaw method, or the like. The type of the yeast to be used for the production of the yeast extract is not particularly limited, and baker's yeast, brewer's yeast, red wine yeast, round yeast, and the like can be used without particular limitation. Among them, it is preferred to use a yeast belonging to the genus Saccharomyces. The yeast extract may be in the form of a paste, a powder or a granule.

In the present invention, among the milk protein and the yeast extract, only the milk protein may be used, and only the yeast extract may be used. Preferably, only the milk protein is used, and more preferably the milk protein and the yeast extract are used in combination.

Here, the mixing ratio at the time of use is preferably 0.1 to 2 parts by mass, more preferably 0.3 to 1 part by mass, based on 1 part by mass of the milk protein.

In the preparation of the component (C), the content ratio of the glycerin of the above (1), the milk protein of (2), and/or the yeast extract is 1 part by mass based on the glycerin of the above (1), and the above (2) The milk protein and/or yeast extract is preferably 0.1 to 3 parts by mass, more preferably 0.1 to 1 part by mass, as the solid content.

Vitamin B12 is used in the formulation of the above component (C). Vitamin B12 is a general name for vitamins containing cobalt. It is a kind of water-soluble vitamin. It has hydroxycobalt vitamin, adenosine cobalt vitamin, methyl cobalt vitamin, cyan cobalt vitamin, cobalt sulfite vitamin, etc. Any of the present invention can be used. By.

In the present invention, it is also possible to use a refined product, and a large amount of food containing vitamin B12 may be used. For example, vitamin B12 is abundantly contained in the liver of seaweed, shellfish, and animal foods.

In the preparation of the component (C), the content ratio of the glycerin of the above (1) and the vitamin B12 of (3) is preferably the vitamin B12 of the above (3) with respect to 1 part by mass of the glycerin of the above (1). It is 0.00001 to 0.001 parts by mass, more preferably 0.00002 to 0.0001 parts by mass.

The decomposition accelerator of the volatile organic halogen compound of the present invention may contain the components other than the above components (A), (B) and (C).

The other component may be, for example, a potassium compound such as glucose, fructose, ammonium sulfate, urea, ammonium salt, sulfur compound, phosphorus compound or potassium chloride which is a source of microbial nutrients, a magnesium compound such as magnesium chloride or magnesium sulfate, or yeast extract. It is also possible to use a substance or a peptone at the same time. Further, the decomposition accelerator of the present invention may be added in an appropriate amount as a decomposition accelerator composition. When the composition of the decomposition accelerator is used, the amount of each additive is not particularly limited. For example, when the powdery yeast extract or fructose is used, the solid content is 1% for the solid content of the extract of the fruit. 200 parts by mass is more preferably more preferably 10 to 100 parts by mass.

However, when the component (C) is used as the present invention, the other components are not contained as components capable of becoming a microbial nutrient component, for example, glucose, fructose, ammonium sulfate, urea, ammonium salt, sulfur compound, phosphorus compound, and chlorination. A potassium compound such as potassium, a magnesium compound such as magnesium chloride or magnesium sulfate, or peptone is preferred. In particular, in the case where there is a sulfuric acid reducing bacteria in the soil or the groundwater, the sulfuric acid ions coexist in competition with the sulfuric acid regenerating bacteria, and thus the decomposition of the volatile organic halogen compound is not performed, so that sulfuric acid such as ammonium sulfate or magnesium sulfate is not contained. Salt is preferred.

The form of the decomposition accelerator of the volatile organic halogen compound of the present invention is not particularly limited, and various forms such as a solid (including powder or granules) or a liquid (including a paste) can be used. Further, it can be used in a state of being diluted with a solvent such as water.

The decomposition accelerator of the present invention promotes decomposition of the volatile organic halogen compound by microorganisms by contact with soil, ground water, and other samples contaminated with the volatile organic halogen compound. The volatile organic halogen compound to be the object of the present invention is preferably an organochlorine-based compound, and examples thereof include carbon tetrachloride, chloroform, dichloromethane, methyl chloride, 1,2-dichloroethane, and 1,1. -dichloroethylene, cis-1,2-dichloroethylene, trans-1,2-dichloroethylene, 1,3-dichloropropene, tetrachloroethylene, 1,1,1-trichloroethane, 1, 1,2-trichloroethane, trichloroethylene, vinyl chloride, and the like.

Among them, the decomposition accelerator of the present invention can suitably promote decomposition of tetrachloroethylene, trichloroethylene, dichloroethylene, vinyl chloride, and the like vinyl chloride.

For example, tetrachloroethylene is decomposed by microorganisms in the order of trichloroethylene, dichloroethylene, monochloroethylene (vinyl chloride), and ethylene.

The decomposition accelerator of the present invention can promote the decomposition of the volatile organic halogen compound by microorganisms, and the microorganism which is originally present in the soil or groundwater to be purified can be used, and the microorganism which is useful for decomposing the volatile organic halogen compound can also be used. Further, it is also possible to use a composition containing such a microorganism at the same time. In other words, when the microorganism or the groundwater to be purified contains a microorganism which sufficiently decomposes the volatile organic halogen compound, the decomposition accelerator or the decomposition accelerator composition of the present invention can be directly applied to the target soil. On the other hand, when the amount of microorganisms in the soil is small, or when it is desired to rapidly decompose, a decomposition accelerator or a decomposition accelerator composition of the present invention containing a composition of microorganisms or microorganisms prepared in advance may be used.

Useful as a decomposition of volatile organic halogen compounds The organism is preferably an anaerobic microorganism, and examples thereof include a microorganism of the genus Clostridium, Dehalobacter, Dehalococcoides, Dehalospirilum, Desulfobacterium, Desulfomonas, and Desulfomonile.

When the decomposition accelerator of the present invention is used, the anaerobic microorganism in the sample containing the volatile organic halogen compound is measured in advance, and for example, the amount of the bacterium belonging to the genus Dehalococcoides is preferably present. The method of quantifying the genus of the genus Dehalococcoides can be carried out by a conventional method such as an instant PCR method (for example, refer to Non-Patent Document 1).

The method for contacting the decomposition accelerating agent of the present invention with soil and/or groundwater is not particularly limited, and the "factory treatment" for treating contaminated soil or groundwater in other places may also be used to purify the contaminated site. The "original purification" of the soil or groundwater is also possible, but it is better to purify it in the original position in view of the maximum effect under the condition that the microorganisms decomposing the volatile organic halogen compound are anaerobic.

In the case of facility-type purification, the method of contacting the decomposition accelerator of the present invention with the soil and/or groundwater in which the microorganisms are present is not particularly limited, and for example, a method of directly injecting the contaminated soil through the excavation stack, and the contaminated soil can be exemplified. A method of mixing and stirring, a method of adding water to a contaminated soil, and adding a liquid to a liquid.

In the case of the original position purification, the method of contacting the decomposition accelerator of the present invention with the soil and/or groundwater in which the microorganism is present is not particularly limited, and for example, a method of directly burying in the soil, injection into the groundwater using an injection well, or Direct injection in soil, or use of underground A method of purifying the wall by a permeation reaction of water flow is also possible, but a direct injection method is preferred.

In addition, the supply amount of the decomposition accelerator of the present invention may be sufficient to obtain a sufficient purification effect, and it is preferable to confirm the range of the contaminated area, the degree of contamination, the type of the contaminant, and the like by prior investigation.

[Examples] <Decomposition Experiment 1 of Vinyl Chloride> [Manufacture of decomposition accelerator 1] <Manufacture of citrus extracts> [Manufacturing Example 1]

The Wenzhou mandarin was thoroughly washed with water, peeled, and the peel (dry weight: 100 g) was pulverized by a disc grinder, and then extracted with 2000 ml of 60 ° C warm water for 1 hour. This was filtered, and the filtrate was concentrated by a rotary evaporator, and dried in a vacuum dryer to obtain a warm water extract (about 10 g) of the peel of Wenzhou mandarin. The obtained peel warm water extract was directly used as the decomposition accelerator A of the present invention.

[Manufacturing Example 2]

To 100 parts by mass of the peeled warm water extract obtained in Production Example 1, 50 parts by mass of the powdery yeast extract and 50 parts by mass of fructose were added and mixed to homogeneity, which was used as the decomposition accelerator B of the present invention.

[Manufacturing Example 3]

The decomposition accelerator C of the present invention was obtained in the same manner as in Production Example 1 except that the hot water of 98 ° C was used instead of the warm water of 60 ° C used for the extraction.

[Manufacturing Example 4]

The decomposition accelerator D of the present invention was obtained in the same manner as in Production Example 1 except that the evening scented orange was used instead of the citrus.

[Manufacturing Example 5]

The decomposition accelerator E of the present invention was obtained in the same manner as in Production Example 1 except that grapefruit was used instead of Wenzhou mandarin.

[Manufacturing Example 6]

The decomposition accelerator F of the present invention was obtained in the same manner as in Production Example 1 except that the lemon was used instead of the mandarin.

[Evaluation method 1 of decomposition accelerator] <Production of Bacterial Liquid> (About decomposition accelerator A~F)

Take 50 mL of the mineral base medium indicated below to 0.1 g/L of the yeast extract medium to a 100 mL capacity glass vial, replace it with nitrogen, and sterilize it in an autoclave, then add 25 mL of vinyl chloride. The groundwater taken from the contaminated soil was replaced with nitrogen, and then 2.5 mL of hydrogen and 0.58 μL of cis-1,2-dichloroethylene (corresponding to 10 mg/L) were sealed and allowed to stand in the dark at 20 °C. Regular determination of vinyl chloride in the headspace For the class, 1 mL was taken at the time when the vinyl chloride was not detected, and inoculation was continued to 75 mL of the autoclaved mineral basal medium supplemented with 0.1 g/L of yeast extract. This subculture was carried out six times as a "bacterial liquid" and used in the following decomposition experiments of vinyl chloride.

<Manufacture of mineral basic medium (pH 7.0~7.5)>

10 ml of the following Salt stock solution, 1 ml of the following Trace element solution A, 1 ml of the following Trace element solution B, 50 μl of azurol sodium solution (0.5% w/v), 0.1 g of sodium acetate, 0.3 g of L-cysteamine The hydrochloride-water mixture, 2.52 g of sodium hydrogencarbonate, 0.048 g of sodium sulfide, and water were filled up to 1000 ml as a mineral base medium.

<Manufacture of Salt stock solution>

The following ingredients were filled in water to 1000 m as a Salt stock solution.

100g NaCl, 50g MgCl ₂ ‧6H ₂ O, 20g KH ₂ PO ₄ , 30g NH ₄ Cl, 30g KCl, 1.5g CaCl ₂ ‧2H ₂ O

<Manufacture of Trace element solution A>

The following ingredients were filled in water to 1000 ml as Trace element solution A.

10 mL HCl (25% solution, w/w), 1.5 g FeCl ₂ ‧4H ₂ O, 0.19 g CoCl ₂ ‧6H ₂ O, 0.1 g MnCl ₂ ‧4H ₂ O, 70 mg Zn Cl ₂ , 6 mg H ₃ BO ₃ , 36 mg Na ₂ MoO ₄ ‧2H ₂ O, 24 mg NiCl ₂ ‧6H ₂ O, 2 mg CuCl ₂ ‧2H ₂ O

<Manufacture of Trace element solution B>

The following ingredients were filled in water to 1000 ml as a Trace element solution B.

6 mg Na ₂ SeO ₃ ‧5H ₂ O, 8 mg Na ₂ WO ₄ ‧2H ₂ O, 0.5 g NaOH

<Decomposition Experiment 1 of Vinyl Chloride>

[Example 1]

It is assumed that the groundwater is contaminated with vinyl chloride, and the decomposition test is carried out by the following method.

The above-mentioned mineral base medium 75 ml was used to make a vial of 100 ml capacity in glass, and the above-mentioned decomposition accelerator A was added to 0.1 g/L, and the mixture was replaced by nitrogen, and then sterilized by a high pressure surface bacteria pot. After cooling, 1.5 ml of the above bacterial solution was added, and after nitrogen substitution, cis-1,2-dichloroethylene (c-DCE) was sealed at 10 μg/ml.

This vial was statically cultured at 20 °C. After 0, 3, 10, 18, 24, 36, 45, 49, 59, 66, 75, 84, and 87 days, the cis-1,2-dichloroethylene in the headspace of the vial was determined by gas chromatography. -DCE) content, vinyl chloride (VC) content, ethylene content.

Regarding the experimental results, first, the growth and decline of cis-1,2-dichloroethylene (c-DCE) content, vinyl chloride (VC) content, and ethylene content from 0 to 87 days As shown in Figure 1.

Further, the activation of microorganisms is considered to be an initial stage, and the amount of decrease in the c-DCE content per day until the 18th day is shown in Table 1 as the initial decomposition rate.

[Example 2]

A decomposition test of vinyl chloride was carried out in the same manner as in Example 1 except that 0.2 g/L of the decomposition accelerator B was added instead of 0.1 g/L of the decomposition accelerator A. The results are shown in Fig. 2 and Table 1.

[Comparative Example 1]

The decomposition test of vinyl chloride was carried out in the same manner as in Example 1 except that 0.1 g/L of the decomposition accelerator A was not added. The results are shown in Fig. 3 and Table 1.

[Comparative Example 2]

The decomposition test of vinyl chloride was carried out in the same manner as in Example 2 except that 0.2 g/L of the decomposition accelerator B and 0.05 g/L of the yeast extract and 0.05 g/L of fructose were added, and the results are shown in Fig. 4 and Table 1. .

[Example 3]

A decomposition test of vinyl chloride was carried out in the same manner as in Example 1 except that 0.1 g/L of the decomposition accelerator C was added instead of 0.1 g/L of the decomposition accelerator A. However, the sampling was performed on the 0th, 3rd, 10th, and 18th days, and the amount of decrease in the c-DCE content per day from the 18th day was calculated in the same manner as in the first embodiment. The decomposition speed is shown in Table 1.

[Example 4]

A decomposition test of vinyl chloride was carried out in the same manner as in Example 1 except that 0.1 g/L of the decomposition accelerator D was added instead of 0.1 g/L of the decomposition accelerator A. Further, the initial decomposition rate was calculated by sampling in the same manner as in Example 3. The results are shown in Table 1.

[Example 5]

A decomposition test of vinyl chloride was carried out in the same manner as in Example 1 except that 0.1 g/L of the decomposition accelerator E was added instead of 0.1 g/L of the decomposition accelerator A. Further, the initial decomposition rate was calculated by sampling in the same manner as in Example 3. The results are shown in Table 1.

[Example 6]

A decomposition test of vinyl chloride was carried out in the same manner as in Example 1 except that 0.1 g/L of the decomposition accelerator F was added instead of 0.1 g/L of the decomposition accelerator A. Further, the initial decomposition rate was calculated by sampling in the same manner as in Example 3. The results are shown in Table 1.

From the results of Comparative Example 1 (Fig. 3), it is apparent that even in the case of the basic medium, even if c-DCE remains on the 87th day, VC also has a tendency to rise, and no occurrence of ethylene is observed at all. The decomposition has hardly been carried out.

On the other hand, from the results of Example 1 (Fig. 1) and Example 2 (Fig. 2), it is apparent that in the sample to which the decomposition accelerator of the present invention is added, c-DCE is almost decomposed on the 50th day, 87th. At the same time, c-DCE and VC are almost completely decomposed.

Further, from the comparison between Example 2 (FIG. 2) and Comparative Example 2 (FIG. 4), it is apparent that the present invention is additionally added to the nutrient component with respect to the sample of the yeast extract and fructose using the conventional nutrient. The decomposition accelerator makes c-DCE decompose more rapidly, especially at the 18th to 50th day. Similarly, the day when the VC maximum concentration was changed from the 59th day (Comparative Example 2, FIG. 4) to the 24th day (Example 2, FIG. 2), it was confirmed that the decomposition accelerator of the present invention was additionally added to the conventional nutrient. The initial decomposition speed can be greatly improved.

Further, as is apparent from Table 1, by the addition of the decomposition accelerator of the present invention, the initial decomposition rate is greatly increased.

Further, although the samples of the yeast extract and fructose using the conventional nutrient have a certain effect of increasing the decomposition rate, it is apparent from the comparison of Examples 1, 3 to 6 and Comparative Example 2 that the decomposition promotion using the present invention is used. One of the agents has a high decomposition promoting effect. Further, from the results of Example 2, the initial decomposition rate was greatly increased by additionally adding the decomposition accelerator of the present invention.

[Example 7 and Comparative Example 3]

It is assumed that the land contaminated with vinyl chloride is subjected to a decomposition test by the following method.

In a screw bag of 1 L capacity, add 700 g of soil taken from land contaminated with trichloroethylene, 300 g of groundwater taken at the same place, and 5 mL of the above bacterial solution, and then extract 0.2 g of decomposition accelerator A and 0.2 g of yeast. The solution was dissolved in 50 mL of distilled water, and after nitrogen substitution, it was left to stand in the dark at room temperature. The concentration of vinyl chloride (trichloroethylene, c-DCE, VC) and the concentration of ethylene in the headspace were measured by gas chromatography at regular intervals. The vinyl chlorides, including trichloroethylene contained in contaminated soil and groundwater, all fell to the environmental reference value after 120 days (trichloroethylene = 0.03 mg / l, dichloroethylene = 0.04 mg / l, VC = 0.002 mg / l) below. The results are shown in Figure 5. Here, in the case where 0.2 g of fructose was substituted for the decomposition accelerator A (Comparative Example 3), even if the groundwater environment reference value or more remained in the VC for 150 days, the land could not be purified. The results are shown in Figure 6.

<Decomposition Experiment 2 of Vinyl Chloride> [Manufacture of decomposition accelerator 2] [Manufacturing Examples 7 to 16]

The glycerin as the component (1), the skim milk powder and/or the yeast extract powder as the component (2), and the vitamin B12 preparation as the component (3) were mixed in accordance with Table 2 to obtain a decomposition accelerator G to P. Further, in the obtained decomposition accelerator, G to N and P are in the form of a paste, and O is in the form of a powder.

Further, the content ratios of (1): (2) and the content ratio of (1): (3) are also shown in Table 2.

The obtained decomposition accelerators G to P were evaluated in accordance with the evaluation method of the decomposition accelerator described below, and the results are shown in Table 2.

[Evaluation method 2 of decomposition accelerator] <Production of Bacterial Liquid>

Take 50 mL of the medium added to the yeast extract in the above mineral base medium to 0.1 g/L to a 100 mL capacity glass vial, replace it with nitrogen, and sterilize it in an autoclave, then add 25 mL of it to be contaminated with vinyl chloride. The groundwater taken from the soil was replaced with nitrogen, and 2.5 mL of hydrogen and 0.58 μL of cis-1,2-dichloroethylene (corresponding to 10 mg/L) were sealed and allowed to stand in the dark at 20 °C. The vinyl chloride in the headspace was periodically measured, and 1 mL was taken at the time when the vinyl chloride was not detected, and then inoculated into 75 mL of an autoclaved mineral basal medium supplemented with 0.1 g/L of yeast extract. Subculture was performed 6 times in this way (only the sixth time without added yeast extract) As a "bacterial liquid", it was used for the decomposition test of the following vinyl chloride.

<Decomposition Experiment 2 of Vinyl Chloride> [Examples 8 to 14 and Comparative Examples 4 to 6]

It is assumed that the groundwater is contaminated with vinyl chloride, and the decomposition test is carried out by the following method.

75 ml of the above mineral base medium was taken to a vial of 100 ml capacity made of glass, and the above decomposition accelerator G~P was added at 0.3 g/L each, and the mixture was replaced with nitrogen, and then sterilized by autoclaving. After cooling, 1.5 ml of the above bacterial solution was added, and after nitrogen substitution, cis-1,2-dichloroethylene (c-DCE) was sealed at 10 μg/ml.

This vial was statically cultured at 20 °C. The c-DCE content, VC content and ethylene content in the headspace of the vial were determined by gas chromatography at regular intervals. As a result of the experiment, the c-DCE content and the VC content are below the groundwater environment reference value, that is, the c-DCE is 0.04 mg/L or less, and the number of days when the VC becomes 0.002 mg/L or less is used as the "required date for decomposition". The number is described in Table 2.

Further, in the case where no decomposition accelerator was added, the same experiment was carried out as Comparative Example 7, and the results are shown in Table 2.

As is clear from the results of Table 2, when the decomposition accelerators of Examples 8 to 14 containing the components (1) to (3) were used, the number of days completely decomposed with respect to the vinyl chloride was 31 days or less, and the component (1) was not contained. Comparative Example 4 was 48 days, Comparative Example 5 containing no (2) component was 45 days, and Comparative Example 6 containing no (3) component was 40 days, and the decomposition rate was extremely low.

Further, when the decomposition accelerator was not used (Comparative Example 7), the decomposition of vinyl chloride was not completed until the 100th day.

Further, in Comparative Examples 9, 11, and 12 using the skim milk powder and/or the yeast extract as the component (2), it is understood that Example 9 using skim milk powder as compared with Example 11 using only the yeast extract. The decomposition rate was higher, and the decomposition rate of Example 12 using skim milk powder and yeast extract was the highest.

Further, in Examples 9 and 13, and 14 in which the contents of the vitamin B12 were changed, it was found that the decomposition rate of the vitamin B12 was not changed in the range of 0.00001 to 0.001 parts by mass based on 1 part by mass of the glycerin.

[Example 15]

It is assumed that the land is contaminated with vinyl chloride, and the decomposition test is carried out by the following method.

In a 1 L capacity screw bottle, put 700 g of soil taken from the land contaminated with trichloroethylene, 300 g of groundwater taken from the same place, add 5 mL of the above bacterial solution, and add 0.5 g of decomposition accelerator dissolved in 50 mL of distilled water. E, substituted by nitrogen, and allowed to stand at room temperature in the dark. The concentration of vinyl chloride (trichloroethylene, c-DCE, VC) and the concentration of ethylene in the headspace were measured by gas chromatography at regular intervals. All vinyl chlorides, including trichloroethylene, contained in contaminated soil and groundwater, all became environmental reference values after 150 days (trichloroethylene = 0.03 mg / l, dichloroethylene = 0.04 mg / l, VC = 0.002 mg / l) below. The results are shown in Figure 7.

<Decomposition Experiment 3 of Vinyl Chloride> [Evaluation method 3 of decomposition accelerator] <Production of Bacterial Liquid>

Take 50ml of the medium added to the yeast extract in the above mineral base medium to 0.1g/L into a 100mL capacity glass vial, replace it with nitrogen, sterilize it in an autoclave, and add 25mL of the quilt. The groundwater taken from the soil contaminated with tetrachloroethylene (PCE) was replaced with nitrogen, and then sealed with 2.5 mL of hydrogen and 0.46 μL (corresponding to 10 mg/L) of PCE, and statically cultured at 20 ° C in the dark. The vinyl chloride in the headspace was periodically measured, and 1 mL was taken at the time when the vinyl chloride was not detected, and the inoculation was continued to 75 mL of the autoclaved mineral basal medium supplemented with 0.1 g/L of the yeast extract. This was subcultured three times as a "bacterial liquid" and used in the following decomposition experiments of vinyl chloride.

[Examples 16 to 18]

It is assumed that the groundwater is contaminated with vinyl chloride, and the decomposition test is carried out by the following method.

75 ml of the above mineral base medium was taken to a 100 ml glass vial, and the decomposition accelerator A (Example 16), the decomposition accelerator G (Example 17), the decomposition accelerator A, and the above were added at 0.2 g/L, respectively. An equal mixture of decomposition accelerator G (Example 18), after being replaced by nitrogen, was sterilized by autoclaving. After cooling, 1.5 ml of the above bacterial solution was added, and after substituting with nitrogen, tetrachloroethylene (PCE) was sealed at 10 μg/ml.

This vial was statically cultured at 20 °C. 0, 3, 10, 18, 24, 36, 45, 49, 59, 66, 75, 84, 87 days after gas chromatography to determine the various vinyl chloride in the top of the vial, namely PCE, TCE, c- DCE, t-DCE, 1,1-DCE, VC content and ethylene content.

Regarding the experimental results, the content of vinyl chloride is below the reference value of the groundwater environment, that is, the PCE is 0.01 mg/L or less, and the TCE becomes 0.03 mg/L or less, cDCE and t-DCE are 0.04 mg/L or less, 1,1-DCE is 0.02 mg/L or less, and the number of days when VC is 0.002 mg/L or less is referred to as "the number of days required for decomposition" Shown in Table 3.

In addition, the amount of reduction in the PCE content up to the 18th day as the initial decomposition rate is shown in Table 3 in consideration of the initial occurrence of the activation of microorganisms.

Claims (6)

A decomposition accelerator for a volatile organic halogen compound, which comprises one or more of the following (A) to (C), (A) warm water to hot water obtained from citrus fruits Extract, (B) extract of warm water to hot water obtained from citrus peel, (C) formulation containing all of the following (1) to (3): (1) glycerin, (2) relative to the foregoing (1) 1 part by mass of glycerin, a milk protein of 0.1 to 3 parts by mass of the solid content, and (3) 0.00001 to 0.001 parts by mass of vitamin B12 based on 1 part by mass of the glycerin of the above (1). A decomposition accelerator composition of a volatile organic halogen compound, which comprises the decomposition accelerator of claim 1 as an active ingredient. A method for promoting decomposition of a volatile organic halogen compound via a microorganism, characterized in that the decomposition accelerator of claim 1 or the decomposition accelerator composition of claim 2 is contacted with soil and/or groundwater containing a volatile organic halogen compound. The method of claim 3, wherein the volatile organic halogen compound is an organochlorine compound. The method of claim 4, wherein the organochlorine-based compound is selected from the group consisting of carbon tetrachloride, chloroform, dichloromethane, methyl chloride, 1,2-dichloroethane, 1,1-dichloroethylene, cis-1. ,2-dichloroethylene, trans-1,2-dichloroethylene, 1,3-dichloropropene, tetrachloroethylene, 1,1,1-trichloroethane, 1,1,2-trichloroethane At least one of a group of trichloroethylene and vinyl chloride. The method of claim 3, wherein the microorganism is selected from the group consisting of Clostridium bacteria, Dehalobacter bacteria, Dehalococcoides bacteria, Dehalospirilum bacteria, and At least one of a group of Desulfobacterium bacteria, Desulfomonas bacteria, and Desulfomonile bacteria.