JP2020110788A - Biodegradation processing method for organic chlorine compound - Google Patents
Biodegradation processing method for organic chlorine compound Download PDFInfo
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- 150000004045 organic chlorine compounds Chemical class 0.000 title claims abstract description 82
- 238000006065 biodegradation reaction Methods 0.000 title claims abstract description 40
- 238000003672 processing method Methods 0.000 title abstract 4
- -1 cyclic ether compound Chemical class 0.000 claims abstract description 25
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 72
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 claims description 65
- 238000000034 method Methods 0.000 claims description 50
- UBOXGVDOUJQMTN-UHFFFAOYSA-N trichloroethylene Natural products ClCC(Cl)Cl UBOXGVDOUJQMTN-UHFFFAOYSA-N 0.000 claims description 41
- XSTXAVWGXDQKEL-UHFFFAOYSA-N Trichloroethylene Chemical group ClC=C(Cl)Cl XSTXAVWGXDQKEL-UHFFFAOYSA-N 0.000 claims description 37
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 29
- 239000002689 soil Substances 0.000 claims description 21
- KFUSEUYYWQURPO-UPHRSURJSA-N cis-1,2-dichloroethene Chemical group Cl\C=C/Cl KFUSEUYYWQURPO-UPHRSURJSA-N 0.000 claims description 14
- 239000010802 sludge Substances 0.000 claims description 10
- WSLDOOZREJYCGB-UHFFFAOYSA-N 1,2-Dichloroethane Chemical compound ClCCCl WSLDOOZREJYCGB-UHFFFAOYSA-N 0.000 claims description 9
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 8
- KNKRKFALVUDBJE-UHFFFAOYSA-N 1,2-dichloropropane Chemical compound CC(Cl)CCl KNKRKFALVUDBJE-UHFFFAOYSA-N 0.000 claims description 7
- UOORRWUZONOOLO-OWOJBTEDSA-N (E)-1,3-dichloropropene Chemical compound ClC\C=C\Cl UOORRWUZONOOLO-OWOJBTEDSA-N 0.000 claims description 6
- 125000004432 carbon atom Chemical group C* 0.000 claims description 6
- 125000001309 chloro group Chemical group Cl* 0.000 claims description 6
- UOORRWUZONOOLO-UHFFFAOYSA-N telone II Natural products ClCC=CCl UOORRWUZONOOLO-UHFFFAOYSA-N 0.000 claims description 6
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 5
- 239000001301 oxygen Substances 0.000 claims description 5
- 229910052760 oxygen Inorganic materials 0.000 claims description 5
- UOCLXMDMGBRAIB-UHFFFAOYSA-N 1,1,1-trichloroethane Chemical compound CC(Cl)(Cl)Cl UOCLXMDMGBRAIB-UHFFFAOYSA-N 0.000 claims description 4
- WNXJIVFYUVYPPR-UHFFFAOYSA-N 1,3-dioxolane Chemical compound C1COCO1 WNXJIVFYUVYPPR-UHFFFAOYSA-N 0.000 claims description 4
- IKZOMJGRWIOEDP-UHFFFAOYSA-N 2-(chloromethyl)-1,3-dioxolane Chemical compound ClCC1OCCO1 IKZOMJGRWIOEDP-UHFFFAOYSA-N 0.000 claims description 4
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 4
- HTWIZMNMTWYQRN-UHFFFAOYSA-N 2-methyl-1,3-dioxolane Chemical compound CC1OCCO1 HTWIZMNMTWYQRN-UHFFFAOYSA-N 0.000 claims description 3
- 238000005086 pumping Methods 0.000 claims description 2
- 241000869142 Pseudonocardia sp. Species 0.000 abstract 1
- LGXVIGDEPROXKC-UHFFFAOYSA-N 1,1-dichloroethene Chemical group ClC(Cl)=C LGXVIGDEPROXKC-UHFFFAOYSA-N 0.000 description 29
- 238000002474 experimental method Methods 0.000 description 28
- 241000894006 Bacteria Species 0.000 description 19
- QERYCTSHXKAMIS-UHFFFAOYSA-N thiophene-2-carboxylic acid Chemical compound OC(=O)C1=CC=CS1 QERYCTSHXKAMIS-UHFFFAOYSA-N 0.000 description 17
- 230000036962 time dependent Effects 0.000 description 17
- 239000000243 solution Substances 0.000 description 12
- BZHJMEDXRYGGRV-UHFFFAOYSA-N Vinyl chloride Chemical group ClC=C BZHJMEDXRYGGRV-UHFFFAOYSA-N 0.000 description 10
- 229910017053 inorganic salt Inorganic materials 0.000 description 10
- 238000005070 sampling Methods 0.000 description 10
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- 238000000354 decomposition reaction Methods 0.000 description 7
- 238000011109 contamination Methods 0.000 description 6
- 125000000816 ethylene group Chemical group [H]C([H])([*:1])C([H])([H])[*:2] 0.000 description 6
- 239000000411 inducer Substances 0.000 description 6
- 150000002894 organic compounds Chemical class 0.000 description 6
- 230000002378 acidificating effect Effects 0.000 description 5
- 230000007423 decrease Effects 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 239000003673 groundwater Substances 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 238000000746 purification Methods 0.000 description 5
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 4
- 238000005273 aeration Methods 0.000 description 4
- 150000004292 cyclic ethers Chemical class 0.000 description 4
- 230000003247 decreasing effect Effects 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- CBENFWSGALASAD-UHFFFAOYSA-N Ozone Chemical compound [O-][O+]=O CBENFWSGALASAD-UHFFFAOYSA-N 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 230000007059 acute toxicity Effects 0.000 description 2
- 231100000403 acute toxicity Toxicity 0.000 description 2
- 238000010923 batch production Methods 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
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- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 238000001878 scanning electron micrograph Methods 0.000 description 2
- 238000011144 upstream manufacturing Methods 0.000 description 2
- 239000002351 wastewater Substances 0.000 description 2
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- XKTYXVDYIKIYJP-UHFFFAOYSA-N 3h-dioxole Chemical compound C1OOC=C1 XKTYXVDYIKIYJP-UHFFFAOYSA-N 0.000 description 1
- 108010077805 Bacterial Proteins Proteins 0.000 description 1
- 206010007269 Carcinogenicity Diseases 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- OTMSDBZUPAUEDD-UHFFFAOYSA-N Ethane Chemical class CC OTMSDBZUPAUEDD-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- RRHGJUQNOFWUDK-UHFFFAOYSA-N Isoprene Chemical compound CC(=C)C=C RRHGJUQNOFWUDK-UHFFFAOYSA-N 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- LFYJSSARVMHQJB-QIXNEVBVSA-N bakuchiol Chemical compound CC(C)=CCC[C@@](C)(C=C)\C=C\C1=CC=C(O)C=C1 LFYJSSARVMHQJB-QIXNEVBVSA-N 0.000 description 1
- 238000009412 basement excavation Methods 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 230000007670 carcinogenicity Effects 0.000 description 1
- 231100000260 carcinogenicity Toxicity 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 150000001805 chlorine compounds Chemical class 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000010924 continuous production Methods 0.000 description 1
- 125000000532 dioxanyl group Chemical group 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 229920001971 elastomer Polymers 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- RTZKZFJDLAIYFH-UHFFFAOYSA-N ether Substances CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 1
- 239000003365 glass fiber Substances 0.000 description 1
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- 231100000086 high toxicity Toxicity 0.000 description 1
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- 239000007924 injection Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical class C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 239000000025 natural resin Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
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- 229910052697 platinum Inorganic materials 0.000 description 1
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- 230000005855 radiation Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Processing Of Solid Wastes (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
- Treatment Of Sludge (AREA)
Abstract
Description
本発明は、有機塩素化合物の構成型１，４−ジオキサン分解菌Ｎ２３株を用いた生分解処理に関する。 The present invention relates to a biodegradation treatment using a constitutive type 1,4-dioxane-degrading bacterium N23 strain of an organochlorine compound.
１，１，１−トリクロロエタン（以下、１，１，１−ＴＣＡともいう）は、様々な有機化合物を溶解することができるため、溶剤、洗浄剤等として広く利用されていた。１，１，１−ＴＣＡは、オゾン層破壊物質の一つとされ、１９９５年に製造が禁止されたが、それ以前に製造されたＴＣＡによる汚染が問題となっている。また、主に、１，１，１−ＴＣＡが使用される前と１，１，１−ＴＣＡが禁止された後においては、トリクロロエチレン（ＴＣＥ）が、溶剤、洗浄剤として使われているが、ＴＣＥによる汚染も１，１，１−ＴＣＡと同様に問題視されている。１，１，１−ＴＣＡ及びＴＣＥは、嫌気性環境下で微生物により、１，１−ジクロロエチレン（以下、１，１−ＤＣＥともいう）やクロロエチレン（以下、ＶＣともいう）等に脱塩素される場合がある。１，１−ＤＣＥ、ＶＣの水への溶解度は、１，１，１−ＴＣＡ（３００ｍｇ／Ｌ水）やＴＣＥ（１，２００ｍｇ／Ｌ水）より大きい（１，１−ＤＣＥ：２，５００ｍｇ／Ｌ水、ＶＣ：８，８００ｍｇ／Ｌ水）。すなわち、１，１，１−ＴＣＡ及びＴＣＥは、脱塩素されるに従って、地下水に溶解して拡散しやすくなるため、汚染範囲が拡大してしまう。 Since 1,1,1-trichloroethane (hereinafter, also referred to as 1,1,1-TCA) can dissolve various organic compounds, it has been widely used as a solvent, a cleaning agent, and the like. 1,1,1-TCA is one of ozone-depleting substances, and its production was banned in 1995, but pollution by TCA produced before that has become a problem. In addition, trichloroethylene (TCE) is mainly used as a solvent and a cleaning agent before 1,1,1-TCA is used and after 1,1,1-TCA is prohibited. Contamination due to TCE is regarded as a problem as in 1,1,1-TCA. 1,1,1-TCA and TCE are dechlorinated into 1,1-dichloroethylene (hereinafter, also referred to as 1,1-DCE), chloroethylene (hereinafter, also referred to as VC), etc. by microorganisms in an anaerobic environment. There are cases where The solubility of 1,1-DCE and VC in water is higher than that of 1,1,1-TCA (300 mg/L water) and TCE (1,200 mg/L water) (1,1-DCE: 2,500 mg/L). L water, VC: 8,800 mg/L water). That is, 1,1,1-TCA and TCE are dissolved in groundwater and easily diffused as they are dechlorinated, so that the contamination range is expanded.
また、１，１，１−ＴＣＡやＴＣＥには、安定剤として、１，４−ジオキサン（以下、ジオキサンともいう）が２〜４％程度添加されていた。ジオキサンは、急性毒性及び慢性毒性を有する上、発がん性も指摘されていることから、日本国では、水道水質基準（０．０５ｍｇ／Ｌ以下）、環境基準（０．０５ｍｇ／Ｌ以下）、排水基準（０．５ｍｇ／Ｌ以下）が設けられている。ジオキサンは、水溶性であるため、水環境中へ放出されると広域に拡散してしまう。
そのため、１，１，１−ＴＣＡ、ＴＣＥの汚染サイトは、１，１，１−ＴＣＡ、ＴＣＥ、１，１−ＤＣＥ、ＶＣ等の有機塩素化合物、ジオキサンによる深刻な複合汚染が発生している場合がある。
Further, 1,4-dioxane (hereinafter, also referred to as dioxane) was added to 1,1,1-TCA and TCE as a stabilizer in an amount of about 2 to 4%. Dioxane has acute toxicity and chronic toxicity, and since carcinogenicity has been pointed out, in Japan, tap water quality standards (0.05 mg/L or less), environmental standards (0.05 mg/L or less), wastewater A standard (0.5 mg/L or less) is provided. Since dioxane is water-soluble, it diffuses over a wide area when released into the aquatic environment.
Therefore, the 1,1,1-TCA and TCE contaminated sites are seriously contaminated by organochlorine compounds such as 1,1,1-TCA, TCE, 1,1-DCE and VC, and dioxane. There are cases.
また、１，１，２−トリクロロエタン（以下、１，１，２−ＴＣＡともいう）は、主に、１，１−ＤＣＥの原料として用いられており、その他の用途として塩素化ゴムの溶剤、油脂、ワックス、天然樹脂などの溶剤、アルカロイドの抽出剤がある。１，１，２−ＴＣＡは、急性毒性及び慢性毒性を持っており、水質汚濁に係る環境基準（０．００６ｍｇ／Ｌ）、地下水の水質汚濁に係る環境基準（０．００６ｍｇ／Ｌ）、土壌汚染に係る環境基準（０．００６ｍｇ／Ｌ）、土壌汚染対策法（土壌溶出量基準０．００６ｍｇ／Ｌ）が設けられている。１，１，２−ＴＣＡは、好気条件での生分解性はないとされている。一方で、嫌気条件では１，１，２−ＴＣＡは分解されるが、より毒性の強いＶＣをはじめ、１，２−ジクロロエタン、１，１−ＤＣＥ、シス−１，２−ジクロロエチレン（以下、シス−１，２−ＤＣＥともいう）等に変換されてしまう。 In addition, 1,1,2-trichloroethane (hereinafter, also referred to as 1,1,2-TCA) is mainly used as a raw material for 1,1-DCE, and as other uses, a solvent for chlorinated rubber, There are oils, waxes, solvents such as natural resins, and extractants for alkaloids. 1,1,2-TCA has acute toxicity and chronic toxicity, and has environmental standards related to water pollution (0.006 mg/L), environmental standards related to water pollution of groundwater (0.006 mg/L), soil. Environmental standards related to pollution (0.006 mg/L) and Soil Contamination Countermeasures Law (soil elution amount standard 0.006 mg/L) are established. 1,1,2-TCA is not biodegradable under aerobic conditions. On the other hand, although 1,1,2-TCA is decomposed under anaerobic conditions, more toxic VC, 1,2-dichloroethane, 1,1-DCE, cis-1,2-dichloroethylene (hereinafter cis -1,2-DCE) and so on.
１，１，１−ＴＣＡ、ＴＣＥ等の有機塩素化合物で汚染された汚染土壌を掘削しないで浄化する方法（原位置浄化）としては、（１）コンプレッサー等で空気を送り揮発させることで除去するスパージング処理。（２）過酸化水素と鉄分を添加してヒドロキシラジカルにより酸化分解させるフェントン法。（３）酸素や栄養剤を送り込んで土壌中の微生物を活性化させて浄化する生物処理（バイオレメディエーション）。等が挙げられる。これらの中では、スパージング処理が一般的であるが、有機塩素化合物は高濃度では揮発するが、低濃度となると揮発性が低下してしまう。 As a method (in-situ purification) of cleaning contaminated soil contaminated with organic chlorine compounds such as 1,1,1-TCA and TCE without excavation (1), air is sent by a compressor etc. to remove it. Sparging treatment. (2) Fenton method in which hydrogen peroxide and iron are added and oxidatively decomposed by hydroxy radicals. (3) Bioremediation, which sends oxygen and nutrients to activate and purify microorganisms in the soil. Etc. Of these, sparging is generally used, but the organochlorine compound volatilizes at high concentrations, but volatility decreases at low concentrations.
一方、１，４−ジオキサンは、上記した浄化方法で水中から除去することが困難であり、過酸化水素を添加してのオゾン処理（Ｏ３／Ｈ２Ｏ２）、紫外線照射下でのオゾン処理（Ｏ３／ＵＶ）、放射線や超音波照射下でのオゾン処理等、複数の物理化学的な酸化方法を併用する促進酸化法においてのみ、処理の有効性が確認されている。しかし、非特許文献１には、１，４−ジオキサン以外の有機物が存在すると、促進酸化法による１，４−ジオキサンの処理効率が低下することが報告されている。 On the other hand, 1,4-dioxane is difficult to remove from water by the above-described purification method, ozone treatment with addition of hydrogen peroxide (O 3 /H 2 O 2 ), ozone under UV irradiation. The effectiveness of the treatment has been confirmed only in the accelerated oxidation method in which a plurality of physicochemical oxidation methods are used in combination, such as treatment (O 3 /UV), ozone treatment under irradiation of radiation or ultrasonic waves. However, Non-Patent Document 1 reports that the presence of an organic substance other than 1,4-dioxane reduces the treatment efficiency of 1,4-dioxane by the accelerated oxidation method.
有機塩素化合物とジオキサンの複合汚染において、有機塩素化合物とジオキサンとを同時に処理することは難しく、有機塩素化合物の処理後に、１，４−ジオキサンを処理する方法が用いられている。しかし、処理化合物ごとに異なる処理方法を採用することは、処理装置、オペレータ、薬品の種類等が増加するため高コストであり、また、処理工程の上流または下流でトラブルが起こると、全ての工程がストップしてしまう。そのため、有機塩素化合物とジオキサンを単一工程で除去可能な技術が求められている。 In the combined pollution of an organic chlorine compound and dioxane, it is difficult to treat the organic chlorine compound and dioxane at the same time, and a method of treating 1,4-dioxane after the treatment of the organic chlorine compound is used. However, adopting a different treatment method for each treatment compound is expensive because the number of treatment devices, operators, types of chemicals, etc. increases, and when trouble occurs upstream or downstream of the treatment process, all processes are treated. Will stop. Therefore, a technique capable of removing the organic chlorine compound and dioxane in a single step is required.
ここで、近年、ジオキサン分解菌を使ったジオキサンの生物処理の検討がされている。有機塩素化合物存在下でのジオキサン分解菌の働きについては、非特許文献２、３に、ジオキサン分解菌であるＣＢ１１９０株が、１，１−ＤＣＥ等の塩素化エチレンを分解できないこと、ＣＢ１１９０株によるジオキサンの分解が、１，１−ＤＣＥ、ｃｉｓ−１，２−ＤＣＥ、ＴＣＥ等の塩素化エチレンにより阻害されることが記載されている。そのため、ＣＢ１１９０株は、ジオキサンと塩素化エチレンの複合汚染に対して、塩素化エチレンを処理できないのはもちろんであるが、塩素化エチレンにより阻害されるためジオキサンを処理することもできない。 Here, in recent years, biological treatment of dioxane using dioxane-degrading bacteria has been studied. Regarding the action of the dioxane-decomposing bacterium in the presence of an organic chlorine compound, Non-Patent Documents 2 and 3 show that the CB1190 strain, which is a dioxane-decomposing bacterium, cannot decompose chlorinated ethylene such as 1,1-DCE. It is described that the decomposition of dioxane is inhibited by chlorinated ethylene such as 1,1-DCE, cis-1,2-DCE and TCE. Therefore, the CB1190 strain cannot of course treat chlorinated ethylene against the combined contamination of dioxane and chlorinated ethylene, but cannot treat dioxane because it is inhibited by chlorinated ethylene.
また、非特許文献４に、有機塩素化合物の分解菌が記載されているが、有機塩素化合物の分解菌は好気条件下では共代謝菌が多く、資化菌、特に、複数の有機塩素化合物を分解できる資化菌は、ほとんど報告されていない。有機塩素化合物による汚染浄化分野で研究されてきた菌は、ほぼ共代謝菌である。しかし、有機塩素化合物が低濃度だと、分解が思うように進まなかったり、トルエンやフェノールといった誘導物質自体が汚染の原因になるという問題がある。他にも有機塩素化合物（ＴＣＥやＤＣＥ）が誘導物質として作用する共代謝菌もいるが、この場合、有機塩素化合物濃度が低くなると、分解速度が低下し、基準値まで下がらない事がある。そのため、有機塩素化合物による汚染浄化は、好気ではなく、嫌気での処理に注目が集まっている。 Further, Non-Patent Document 4 describes a degrading bacterium of an organochlorine compound, but the degrading bacterium of an organochlorine compound is often a co-metabolizing bacterium under aerobic conditions. Few assimilating bacteria capable of decomposing spores have been reported. Bacteria that have been studied in the field of pollution purification with organochlorine compounds are almost co-metabolizing bacteria. However, if the concentration of the organic chlorine compound is low, there are problems that the decomposition does not proceed as expected and that the inducer itself such as toluene or phenol causes pollution. In addition, there are co-metabolites in which an organic chlorine compound (TCE or DCE) acts as an inducer, but in this case, when the concentration of the organic chlorine compound becomes low, the decomposition rate may decrease and the standard value may not be reached. Therefore, the purification of pollution by an organic chlorine compound is not aerobic, and attention is focused on anaerobic treatment.
本発明者らは、特許文献１において、構成型１，４−ジオキサン分解菌であるＮ２３株を報告している。Ｎ２３株は、これまでに報告されている構成型１，４−ジオキサン分解菌の中で、最も高い１，４−ジオキサン最大比分解速度を示し、１，４−ジオキサンを始めとする環状エーテルの生分解に非常に有望である。
また、本発明者らは、特許文献２において、Ｎ２３株は、酸性環境下で活性がほとんど低下しないため、ｐＨ３．０以上５．５以下の酸性環境下での有機化合物の生分解処理に用いることを提案している。
The present inventors have reported in Patent Document 1 the N23 strain, which is a constitutive type 1,4-dioxane-degrading bacterium. The N23 strain shows the highest maximum specific decomposition rate of 1,4-dioxane among the constitutive 1,4-dioxane-degrading bacteria reported so far, and the cyclic ether of 1,4-dioxane and other cyclic ethers Very promising for biodegradation.
In addition, the present inventors use the N23 strain for the biodegradation treatment of an organic compound in an acidic environment of pH 3.0 or more and 5.5 or less in Patent Document 2 because the activity of the N23 strain hardly decreases in the acidic environment. Is proposing that.
有機塩素化合物の好気での生分解処理方法、特に、有機塩素化合物と環状エーテル化合物とを生分解する生分解処理方法を提供することを課題とする。 It is an object of the present invention to provide an aerobic biodegradation treatment method for an organochlorine compound, and particularly to provide a biodegradation treatment method for biodegrading an organochlorine compound and a cyclic ether compound.
本発明の課題を解決するための手段は以下の通りである。
１．受託番号ＮＩＴＥ ＢＰ−０２０３２として寄託されたＮ２３株を、好気環境下で有機塩素化合物と接触させる工程を含むことを特徴とする、有機塩素化合物の生分解処理方法。
２．前記有機塩素化合物が、炭素原子数１〜３、塩素原子数１〜３であることを特徴とする１．に記載の生分解処理方法。
３．前記有機塩素化合物が、ジクロロメタン、１，２−ジクロロエタン、クロロエチレン、１，１−ジクロロエチレン、シス−１，２−ジクロロエチレン、１，２−ジクロロプロパン、１，３−ジクロロプロペン、１，１，２−トリクロロエタン、トリクロロエチレンの１種以上であることを特徴とする１．または２．に記載の生分解処理方法。
４．前記有機塩素化合物と、環状エーテル化合物とを処理することを特徴とする１．〜３．のいずれかに記載の生分解処理方法。
５．前記環状エーテル化合物が、１，４−ジオキサン、１，３−ジオキソラン、２−メチル−１，３−ジオキソラン、２−クロロメチル−１，３−ジオキソラン、テトラヒドロフランの１種以上であることを特徴とする４．に記載の生分解処理方法。
６．前記有機塩素化合物を含む汚染土壌に、前記Ｎ２３株を注入する工程、スパージング処理により酸素を供給する工程を含むことを特徴とする１．〜５．のいずれかに記載の生分解処理方法。
７．地中から前記有機塩素化合物を含む汚染水を揚水する工程、該汚染水を前記Ｎ２３株を用いた標準活性汚泥法により処理する工程を含むことを特徴とする１．〜６．のいずれかに記載の生分解処理方法。
Means for solving the problems of the present invention are as follows.
1. A method for biodegrading an organochlorine compound, comprising the step of contacting the N23 strain deposited under the deposit number NITE BP-02032 with an organochlorine compound under an aerobic environment.
2. 1. The organic chlorine compound has 1 to 3 carbon atoms and 1 to 3 chlorine atoms. Biodegradation treatment method described in.
3. The organic chlorine compound is dichloromethane, 1,2-dichloroethane, chloroethylene, 1,1-dichloroethylene, cis-1,2-dichloroethylene, 1,2-dichloropropane, 1,3-dichloropropene, 1,1,2. -It is one or more of trichloroethane and trichlorethylene. Or 2. Biodegradation treatment method described in.
4. 1. Treating the organochlorine compound and a cyclic ether compound. ~3. The method for biodegradation according to any one of 1.
5. The cyclic ether compound is one or more of 1,4-dioxane, 1,3-dioxolane, 2-methyl-1,3-dioxolane, 2-chloromethyl-1,3-dioxolane, and tetrahydrofuran. Do 4. Biodegradation treatment method described in.
6. 1. A step of injecting the N23 strain into a contaminated soil containing the organochlorine compound, and a step of supplying oxygen by a sparging treatment. ~5. The method for biodegradation according to any one of 1.
7. 1. A method of pumping contaminated water containing the organic chlorine compound from the ground, and a step of treating the contaminated water by a standard activated sludge method using the N23 strain. ~6. The method for biodegradation according to any one of 1.
Ｎ２３株により、有機塩素化合物を好気環境下で生分解処理することができる。特に、炭素原子数１〜３、塩素原子数１〜３の有機塩素化合物は、水への溶解度が大きいもの、毒性が高いものが多いが、Ｎ２３株により、低コストで高い効率で生分解処理することができる。
Ｎ２３株は、複数の有機塩素化合物を生分解することができ、また、事前に誘導物質による馴養が不要である。また、Ｎ２３株は、有機塩素化合物と環状エーテル化合物を同時に生分解することができる。そのため、本発明の生分解処理方法は、１，１，１−ＴＣＡの汚染サイト等、有機塩素化合物と環状エーテル化合物との複合汚染が広がっている汚染サイトに好適に用いることができる。
With the N23 strain, an organic chlorine compound can be biodegraded in an aerobic environment. In particular, many of the organic chlorine compounds having 1 to 3 carbon atoms and 1 to 3 chlorine atoms have high solubility in water and high toxicity, but the N23 strain enables low-cost, high-efficiency biodegradation treatment. can do.
The N23 strain is capable of biodegrading a plurality of organochlorine compounds, and does not require prior acclimation with an inducer. The N23 strain can simultaneously biodegrade an organic chlorine compound and a cyclic ether compound. Therefore, the biodegradation treatment method of the present invention can be suitably used for a polluted site where complex pollution of an organochlorine compound and a cyclic ether compound spreads, such as a polluted site of 1,1,1-TCA.
有機塩素化合物は、その種類により生分解性に差があるが、土壌へのスパージング処理および／または活性汚泥法における曝気処理と、Ｎ２３株による生分解を組み合わせることにより、複数種の有機塩素化合物、環状エーテル化合物を、効率的に同時に処理することができる。 The organochlorine compound has different biodegradability depending on its type, but by combining sparging treatment to soil and/or aeration treatment in the activated sludge method and biodegradation by N23 strain, a plurality of organochlorine compounds, The cyclic ether compound can be efficiently treated simultaneously.
「Ｎ２３株」
Ｎ２３株は、受託番号ＮＩＴＥ ＢＰ−０２０３２として、独立行政法人 製品評価技術基盤機構 特許微生物寄託センター（ＮＰＭＤ）（日本国千葉県木更津市かずさ鎌足２−５−８（郵便番号２９２−０８１８））に、２０１５年４月１０日付で国際寄託されている。Ｎ２３株のＳＥＭ画像を図１に示す。Ｎ２３株は、グラム染色性が陽性、カタラーゼ反応が陽性である。
"N23 shares"
The N23 strain has a consignment number NITE BP-02032, and is an independent administrative agency Product Evaluation Technology Platform Organization, Patent Microorganism Depositary Center (NPMD) (2-5-8 Kazusa Kamasa, Kisarazu City, Chiba Prefecture, Japan (zip code 292-0818)). Has been internationally deposited on April 10, 2015. The SEM image of N23 strain is shown in FIG. The N23 strain has positive Gram stainability and positive catalase reaction.
Ｎ２３株は、これまでに報告されている構成型資化菌の中で最も高い１，４−ジオキサン最大比分解速度を有する構成型１，４−ジオキサン分解菌である（特許文献１）。
Ｎ２３株は、ジオキサンを極低濃度まで分解することができる、高濃度のジオキサンを処理することができる、予め馴養する必要がない、ジオキサンだけでなく、１，３−ジオキソラン、２−メチル−１，３−ジオキソラン、２−クロロメチル−１，３−ジオキソラン、テトラヒドロフラン等の環状エーテル化合物を効率よく分解することができる等、環状エーテル化合物の生分解処理に好適に利用することができる。
The N23 strain is a constitutive type 1,4-dioxane-degrading bacterium having the highest 1,4-dioxane maximum specific decomposition rate among the constitutive assimilating bacteria reported so far (Patent Document 1).
The N23 strain can decompose dioxane to an extremely low concentration, can treat high concentration of dioxane, does not need to be preconditioned, and is not limited to dioxane but also 1,3-dioxolane and 2-methyl-1. , 3-dioxolane, 2-chloromethyl-1,3-dioxolane, tetrahydrofuran, and other cyclic ether compounds can be efficiently decomposed, and can be suitably used for biodegradation treatment of cyclic ether compounds.
本発明者らは、Ｎ２３株によるジオキサン汚染土壌処理に際し、汚染土壌中に有機塩素化合物が存在していてもＮ２３株のジオキサン分解活性が低下しないことを発見した。本発明者らは、Ｎ２３株の有機塩素化合物の生分解性についてさらに研究を進めたところ、Ｎ２３株が、有機塩素化合物を好気環境下で、誘導物質による馴養をすることなく生分解できることを見出し、本発明を完成させるに至った。
すなわち、本発明は、受託番号ＮＩＴＥ ＢＰ−０２０３２として寄託されたＮ２３株を用いる、有機塩素化合物の生分解処理方法に関する。
The present inventors have discovered that, when dioxane-contaminated soil is treated with the N23 strain, the dioxane-degrading activity of the N23 strain is not reduced even if an organochlorine compound is present in the contaminated soil. The present inventors have further researched the biodegradability of the organochlorine compound of the N23 strain, and found that the N23 strain can biodegrade the organochlorine compound in an aerobic environment without acclimatization with an inducer. Heading out, the present invention has been completed.
That is, the present invention relates to a method for biodegrading an organochlorine compound using the N23 strain deposited under the deposit number NITE BP-02032.
「有機塩素化合物」
本発明においてＮ２３株を用いて生分解する有機塩素化合物としては、Ｎ２３株が生分解できる化合物であれば特に限定されず、また、２種以上の有機塩素化合物であってもよい。Ｎ２３株が生分解できる有機塩素化合物としては、例えば、炭素原子数１〜３、塩素原子数１〜３の有機塩素化合物が挙げられ、炭素原子数１〜３、塩素原子数１〜２の有機塩素化合物が好ましい。具体的には、ジクロロメタン、１，２−ジクロロエタン、クロロエチレン（塩化ビニルモノマー）、１，１−ジクロロエチレン、シス−１，２−ジクロロエチレン、１，２−ジクロロプロパン、１，３−ジクロロプロペン、１，１，２−トリクロロエタン、トリクロロエチレン等が挙げられる。これらの中で、特に、１，２−ジクロロプロパン、１，１，２−トリクロロエタンは、本発明者らが調査したところ、これまでに嫌気性微生物による生分解しか報告されておらず、１，２−ジクロロプロパンの好気性微生物による生分解は、Ｎ２３株が世界初の事例である。
"Organic chlorine compounds"
In the present invention, the organochlorine compound biodegradable using the N23 strain is not particularly limited as long as it is a compound capable of biodegrading the N23 strain, and may be two or more types of organochlorine compounds. Examples of the organic chlorine compound that can be biodegraded by the N23 strain include organic chlorine compounds having 1 to 3 carbon atoms and 1 to 3 chlorine atoms, and organic compounds having 1 to 3 carbon atoms and 1 or 2 chlorine atoms. Chlorine compounds are preferred. Specifically, dichloromethane, 1,2-dichloroethane, chloroethylene (vinyl chloride monomer), 1,1-dichloroethylene, cis-1,2-dichloroethylene, 1,2-dichloropropane, 1,3-dichloropropene, 1 , 1,2-trichloroethane, trichloroethylene and the like. Among these, in particular, 1,2-dichloropropane and 1,1,2-trichloroethane were investigated by the present inventors and only biodegradation by an anaerobic microorganism has been reported so far. The N23 strain is the world's first case of biodegradation of 2-dichloropropane by an aerobic microorganism.
「環状エーテル化合物」
Ｎ２３株は、環状エーテル化合物の有無に関わらず、有機塩素化合物を生分解することができる。そのため、Ｎ２３株は、有機塩素化合物のみの生分解処理、有機塩素化合物と環状エーテル化合物の両方の生分解処理に用いることができる。本発明において有機塩素化合物とともに生分解する環状エーテル化合物としては、Ｎ２３株が生分解できるものであれば特に限定されない。また、有機塩素化合物と環状エーテル化合物の両方を生分解する場合、有機塩素化合物、環状エーテル化合物のいずれか、または両方が、２種以上であってもよい。Ｎ２３株が生分解できる環状エーテル化合物としては、例えば、１，４−ジオキサン、１，３−ジオキソラン、２−メチル−１，３−ジオキソラン、２−クロロメチル−１，３−ジオキソラン、テトラヒドロフラン等を挙げることができる。なお、複数の有機化合物を生分解する場合、Ｎ２３株は、炭素源として利用しやすいものから順に生分解するため、存在する有機化合物の種類や量により、各有機化合物の生分解終了までの時間は異なる。
"Cyclic ether compound"
N23 strain can biodegrade organic chlorine compounds with or without cyclic ether compounds. Therefore, the N23 strain can be used for the biodegradation treatment of only the organochlorine compound and the biodegradation treatment of both the organochlorine compound and the cyclic ether compound. In the present invention, the cyclic ether compound that is biodegradable together with the organic chlorine compound is not particularly limited as long as it can biodegrade the N23 strain. When biodegrading both the organochlorine compound and the cyclic ether compound, two or more of the organochlorine compound and/or the cyclic ether compound may be used. Examples of the cyclic ether compound that can be biodegraded by the N23 strain include 1,4-dioxane, 1,3-dioxolane, 2-methyl-1,3-dioxolane, 2-chloromethyl-1,3-dioxolane, and tetrahydrofuran. Can be mentioned. In addition, when biodegrading a plurality of organic compounds, the N23 strain biodegrades in order from the one that is easy to use as a carbon source. Is different.
「生分解処理」
本発明の有機塩素化合物の生分解処理方法は、Ｎ２３株を、好気環境下で有機塩素化合物と接触させる工程を含むことを特徴とする。本発明により生分解処理を行う処理対象は、有機塩素化合物を含むものであれば特に制限されないが、具体的には、不法廃棄現場等の汚染サイト、工場跡地、産業廃棄物処理場等の汚染土壌や、工場排水、一般下水、汚染サイト近辺の地下水等の汚染水等が挙げられる。また、処理対象は、有機塩素化合物と環状エーテル化合物の両方を含むものでもよい。
Ｎ２３株は、培養液からろ別した菌体、凍結保存した菌体、乾燥保存した菌体、凍結乾燥した菌体、Ｎ２３株を樹脂等に固定化した固定化担体、あるいは培養液やその濃縮液等のＮ２３株を含む懸濁液等の任意の形態で生分解処理に用いることができる。
"Biodegradation treatment"
The method for biodegrading an organochlorine compound of the present invention is characterized by including a step of contacting the N23 strain with an organochlorine compound under an aerobic environment. The subject to be subjected to biodegradation treatment according to the present invention is not particularly limited as long as it contains an organic chlorine compound. Examples include soil, contaminated water such as factory wastewater, general sewage, and groundwater near the contaminated site. Further, the treatment target may include both the organic chlorine compound and the cyclic ether compound.
The N23 strain is a bacterial cell separated by filtration from a culture solution, a cryopreserved bacterial cell, a dry-preserved bacterial cell, a freeze-dried bacterial cell, an immobilized carrier in which the N23 strain is immobilized on a resin, or a culture solution or its concentration It can be used for biodegradation treatment in any form such as a suspension containing N23 strain such as liquid.
Ｎ２３株による生分解処理時のｐＨは、Ｎ２３株が活動可能なｐＨであれば特に制限されないが、ｐＨ３．０以上５．５以下であることが好ましい。特許文献２に記載したように、Ｎ２３株は、酸性環境下であっても、ほとんど活動性が低下しない。それに対し、一般的な雑菌は、中性環境下が至適ｐＨであるため、ｐＨ３．０以上５．５以下の酸性環境下では、活動が抑制される。そのため、ｐＨ３．０以上５．５以下の酸性環境下では、雑菌の繁殖（コンタミネーション）を抑制することができ、Ｎ２３株による生分解処理を効率的に行うことができる。 The pH at the time of biodegradation treatment with the N23 strain is not particularly limited as long as the N23 strain can be activated, but it is preferably pH 3.0 or more and 5.5 or less. As described in Patent Document 2, the N23 strain shows almost no decrease in activity even in an acidic environment. On the other hand, since general miscellaneous bacteria have an optimum pH in a neutral environment, their activities are suppressed in an acidic environment of pH 3.0 or more and 5.5 or less. Therefore, in an acidic environment of pH 3.0 or more and 5.5 or less, it is possible to suppress the propagation of contaminants (contamination) and efficiently perform biodegradation treatment with the N23 strain.
Ｎ２３株により、汚染土壌中の有機塩素化合物を生分解処理することができる。Ｎ２３株を用いた汚染土壌中の有機塩素化合物の生分解処理方法は特に制限されず、公知の方法を採用することができる。例えば、汚染土壌にＮ２３株を加えて混合撹拌する、汚染土壌中に懸濁液等の形態でＮ２３株を注入する等の方法を挙げることができる。汚染土壌にＮ２３株を注入する場合、Ｎ２３株は、好気性細菌であるため、Ｎ２３株を注入する工程、スパージングにより土壌中に酸素を供給する工程を含むことが好ましい。また、一般に土壌中には栄養素が不足しているため、汚染土壌中に炭素源、無機塩類等を注入する工程を含むことが好ましい。スパージングを行うことにより、土壌中の有機塩素化合物を揮発させて低濃度とすることができる。そして、スパージングでは取り除けない低濃度の有機塩素化合物をＮ２３株により生分解処理することができるため、浄化処理に必要な時間を短縮することができる。 The N23 strain can biodegrade organic chlorine compounds in contaminated soil. The biodegradation treatment method of the organochlorine compound in the contaminated soil using N23 strain is not particularly limited, and a known method can be adopted. For example, the N23 strain may be added to the contaminated soil and mixed and stirred, or the N23 strain may be injected into the contaminated soil in the form of a suspension or the like. When the N23 strain is injected into the contaminated soil, since the N23 strain is an aerobic bacterium, it is preferable to include the step of injecting the N23 strain and the step of supplying oxygen into the soil by sparging. Further, since nutrients are generally insufficient in soil, it is preferable to include a step of injecting a carbon source, inorganic salts and the like into contaminated soil. By performing the sparging, the organic chlorine compound in the soil can be volatilized to a low concentration. Then, since the low-concentration organochlorine compound that cannot be removed by sparging can be biodegraded by the N23 strain, the time required for the purification treatment can be shortened.
Ｎ２３株により、汚染水中の有機塩素化合物を生分解処理することができる。Ｎ２３株を用いた汚染水中の有機塩素化合物の生分解処理方法は特に制限されず、公知の方法を採用することができる。例えば、標準活性汚泥法における処理方法を挙げることができる。標準活性汚泥法は、曝気槽に、固定化担体や懸濁液等の形態でＮ２３株を加えるだけで有機塩素化合物を処理することができるため、従来の標準活性汚泥法で用いられる設備をほとんどそのまま活用することができる。また、標準活性汚泥法における曝気により、汚染水中の有機塩素化合物を揮発させて取り除くことができる。 N23 strain can biodegrade organic chlorine compounds in contaminated water. The biodegradation treatment method of the organic chlorine compound in the contaminated water using the N23 strain is not particularly limited, and a known method can be adopted. For example, a treatment method in the standard activated sludge method can be mentioned. Since the standard activated sludge method can treat organochlorine compounds by simply adding N23 strain in the form of immobilized carrier or suspension to the aeration tank, most of the equipment used in the conventional standard activated sludge method can be treated. It can be used as it is. Further, by aeration in the standard activated sludge method, the organic chlorine compound in the contaminated water can be volatilized and removed.
標準活性汚泥法における処理は、（１）汚染水のＮ２３株による生分解処理工程、（２）Ｎ２３株を含む活性汚泥や担体等を沈殿させ、処理後の上澄みを排水する排水工程、（３）新たな汚染水を投入する汚染水投入工程を、（１）→（２）→（３）→（１）→・・・と、この順で繰り返す、いわゆるフェッドバッチプロセス、上流での汚染水の投入と下流での処理水の排水とを同量で連続的に行う工程を有する連続プロセスのいずれの方法でも行うことができる。フェッドバッチプロセスは、曝気槽での初期有機化合物（汚染物）濃度が高いため、生分解処理速度を高く保つことができ、また、排水時にＮ２３株の流出が少なく、プロセスを繰り返す毎にＮ２３株量が増加して１回のプロセスに必要な時間を徐々に短くすることができる。 The treatment in the standard activated sludge method includes (1) a biodegradation treatment step of N23 strain of contaminated water, (2) a drainage step of precipitating activated sludge and carrier containing N23 strain, and draining the supernatant after treatment, (3) ) The so-called fed-batch process, the contaminated water at the upstream, is repeated in the order of (1)→(2)→(3)→(1)→... Can be carried out by any method of a continuous process having a step of continuously supplying the same amount of water and discharging the treated water in the downstream. In the fed-batch process, the concentration of initial organic compounds (pollutants) in the aeration tank is high, so the biodegradation treatment rate can be kept high, and the N23 strain does not flow out a lot during drainage, and the N23 strain is repeated every time the process is repeated. The amount can be increased to gradually reduce the time required for a single process.
有機塩素化合物の中で、ジクロロメタン（１３，２００ｍｇ／Ｌ水）、１，２−ジクロロエタン（８，６００ｍｇ／Ｌ水）、クロロエチレン（８，８００ｍｇ／Ｌ水）、シス−１，２−ジクロロエチレン（５，１００ｍｇ／Ｌ水）、１，２−ジクロロプロパン（２，８００ｍｇ／Ｌ水）、１，３−ジクロロプロペン（２，０００ｍｇ／Ｌ水）は、水への溶解度が高い。また、エーテル結合による極性を有する環状エーテルは、水に易溶解性である。そのため、有機塩素化合物による汚染サイト、または、有機塩素化合物と環状エーテルによる汚染サイトでは、処理対象である有機化合物が地下水に溶解して汚染範囲が広範囲に亘る場合がある。汚染が広範囲に亘る場合は、汚染土壌へのＮ２３株の注入と、揚水した地下水（汚染水）の標準活性汚泥法による生分解処理の両方を行うことが、汚染の拡大を防ぎ、また、処理期間を短くすることができるため好ましい。この際、スパージングを行うことが、Ｎ２３株への酸素を供給するのみならず、水への溶解度が低く揮発性の高い有機塩素化合物を取り除くことができるため、好ましい。 Among the organic chlorine compounds, dichloromethane (13,200 mg/L water), 1,2-dichloroethane (8,600 mg/L water), chloroethylene (8,800 mg/L water), cis-1,2-dichloroethylene ( 5,100 mg/L water), 1,2-dichloropropane (2,800 mg/L water), and 1,3-dichloropropene (2,000 mg/L water) have high solubility in water. Further, the cyclic ether having polarity by an ether bond is easily soluble in water. Therefore, at the site polluted with the organic chlorine compound or the site polluted with the organic chlorine compound and the cyclic ether, the organic compound to be treated may be dissolved in the groundwater and the contamination range may be wide. When the pollution is widespread, both injection of N23 strain into the contaminated soil and biodegradation of pumped groundwater (contaminated water) by the standard activated sludge method prevent the spread of pollution and This is preferable because the period can be shortened. At this time, sparging is preferable because not only oxygen can be supplied to the N23 strain but also an organic chlorine compound having low solubility in water and high volatility can be removed.
「Ｎ２３株」
３００ｍＬ容量のバッフル付の三角フラスコにＭＧＹ培地（Ｍａｌｔ Ｅｘｔｒａｃｔ：１０ｇ／Ｌ、グルコース：４ｇ／Ｌ、Ｙｅａｓｔ Ｅｘｔｒａｃｔ：４ｇ／Ｌ、ｐＨ７．３）を１００ｍＬ添加し、オートクレーブにて滅菌処理（１２１℃、１５分）を行った。その後、Ｎ２３株を一白金耳で植菌し、回転振盪培養（２８℃、１２０ｒｐｍ）を７日間行った（前々培養）。
培養後、ＭＧＹ培地に植え継ぎ、同様の条件にて培養を行った（前培養）。
前培養にて得られた培養液を遠心分離によって集菌・回収し、無機塩培地（組成：１ｇ／Ｌ Ｋ２ＨＰＯ４、１ｇ／Ｌ （ＮＨ４）２ＳＯ４、５０ｍｇ／Ｌ ＮａＣｌ、２００ｍｇ／Ｌ ＭｇＳＯ４・７Ｈ２Ｏ、１０ｍｇ／Ｌ ＦｅＣｌ３、５０ｍｇ／Ｌ ＣａＣｌ２、ｐＨ：７．３）を加えて、菌体の洗浄を行った。洗浄後の菌体を、無機塩培地で懸濁したものを植菌液とした。
"N23 shares"
100 mL of MGY medium (Malt Extract: 10 g/L, glucose: 4 g/L, Yeast Extract: 4 g/L, pH 7.3) was added to a 300 mL capacity baffled Erlenmeyer flask and sterilized by autoclave (121° C., 121° C.). 15 minutes). Then, the N23 strain was inoculated with one platinum loop and cultivated with rotary shaking (28° C., 120 rpm) for 7 days (pre-preculture).
After culturing, the cells were subcultured in MGY medium and cultivated under the same conditions (preculture).
The culture solution obtained in the pre-culture was collected and collected by centrifugation to obtain an inorganic salt medium (composition: 1 g/L K 2 HPO 4 , 1 g/L (NH 4 )2SO 4 , 50 mg/L NaCl, 200 mg/ L MgSO 4 · 7H 2 O, 10mg / L FeCl 3, 50mg / L CaCl 2, pH: 7.3) was added and was washed bacterial cells. The washed bacterial cells were suspended in an inorganic salt medium and used as an inoculum.
「分析方法」
・濃度測定
１，４−ジオキサン及び有機塩素化合物は、ヘッドスペースガスクロマトグラフィー質量分析装置（島津製作所社製、ＧＣ／ＭＳ ＱＰ−２０１０ ｐｌｕｓ）を用いてＪＩＳ Ｋ０１２５に準じて行った。
"Analysis method"
-Concentration measurement 1,4-dioxane and an organic chlorine compound were measured according to JIS K0125 using a headspace gas chromatography mass spectrometer (manufactured by Shimadzu Corporation, GC/MS QP-2010 plus).
・菌体濃度
Ｎ２３株の菌体タンパク濃度は、既報（Meyers et al., Novel method for rapid measurement of growth of mycobacteria in detergent-free media, J. Clin. Microbiol.,36 (9) 2752〜2754 (1998)）に準じて測定した。
Ｎ２３株の乾燥菌体重量及び微生物濃度は、ガラス繊維濾紙ＧＦ／Ｂ（粒子保持能 １．０μｍ、Ｗｈａｔｍａｎ）を用いて試料をろ過し、１０５℃にて２時間乾燥した後の重量から、ろ過前のフィルター重量を差し引いて求めた。
-Bacterial cell concentration The bacterial protein concentration of the N23 strain has been reported previously (Meyers et al., Novel method for rapid measurement of growth of mycobacteria in detergent-free media, J. Clin. Microbiol., 36 (9) 2752-2754 ( 1998)).
The dry cell weight and microbial concentration of the N23 strain were determined by filtering the sample using glass fiber filter paper GF/B (particle retention capacity 1.0 μm, Whatman) and drying it at 105° C. for 2 hours. It was determined by subtracting the weight of the previous filter.
「実験１」
１２０ｍｌ容のバイアル瓶にＮ２３株（菌体濃度：５００ｍｇ−ｄｒｙ ｃｅｌｌ／Ｌ）、１，４−ジオキサン（１．０ｍｇ／Ｌ）、１，１−ＤＣＥ（１．０、５．０、１０．０ｍｇ／Ｌ）、無機塩培地を条件毎に合計で２４ｍＬとなるように添加した。その後、ブチルゴム栓で密栓し３０℃、１２０ｒｐｍの条件で振盪しながら試験を行った（Ｎ＝２）。
実験は下記５つの条件を用意して行った。
条件１：Ｎ２３株及び１，１−ＤＣＥを加えない（コントロール）
条件２：１，１−ＤＣＥを加えない（ポジティブコントロール）
条件３：１，１−ＤＣＥ濃度が１．０ｍｇ／Ｌ
条件４：１，１−ＤＣＥ濃度が５．０ｍｇ／Ｌ
条件５：１，１−ＤＣＥ濃度が１０．０ｍｇ／Ｌ
"Experiment 1"
N23 strain (cell concentration: 500 mg-dry cell/L), 1,4-dioxane (1.0 mg/L), 1,1-DCE (1.0, 5.0, 10. 0 mg/L), and an inorganic salt medium was added so that the total amount was 24 mL for each condition. Then, the test was carried out while sealing with a butyl rubber stopper and shaking at 30° C. and 120 rpm (N=2).
The experiment was conducted under the following five conditions.
Condition 1: N23 strain and 1,1-DCE are not added (control)
Condition 2: 1,1,1-DCE is not added (positive control)
Condition 3: 1,1-DCE concentration is 1.0 mg/L
Condition 4: 1,1-DCE concentration is 5.0 mg/L
Condition 5: 1,1-DCE concentration is 10.0 mg/L
培養開始３日後、４日後、５日後にサンプリングを行い、溶液中の１，１−ＤＣＥ、１，４−ジオキサン濃度を測定した。
各条件の１，４−ジオキサン濃度及び１，１−ＤＣＥ濃度の経時変化を図２、３に示す。なお、数値は２連の平均値を用いている。
Sampling was performed 3 days, 4 days, and 5 days after the start of culture, and the 1,1-DCE and 1,4-dioxane concentrations in the solution were measured.
The changes over time in the 1,4-dioxane concentration and the 1,1-DCE concentration under each condition are shown in FIGS. In addition, the numerical value uses the average value of 2 consecutive.
Ｎ２３株が、１，１−ＤＣＥを生分解できることが確かめられた。
Ｎ２３株は、ＣＢ１１９０株ではジオキサン分解を最も強く阻害する１，１−ＤＣＥの存在下でも、１，４−ジオキサンが分解できることが確かめられ、さらに、ジオキサンよりも先に１，１−ＤＣＥを分解することが確認できた。１，１−ＤＣＥ濃度が５．０ｍｇ／Ｌ以下程度であれば、１，４−ジオキサンを問題なく環境基準値まで分解すると共に１，１−ＤＣＥも分解できることが確認できた。
It was confirmed that the N23 strain can biodegrade 1,1-DCE.
It was confirmed that the N23 strain can decompose 1,4-dioxane even in the presence of 1,1-DCE, which strongly inhibits dioxane decomposition in the CB1190 strain, and further decomposes 1,1-DCE before dioxane. I was able to confirm that It was confirmed that when the 1,1-DCE concentration was about 5.0 mg/L or less, 1,4-dioxane could be decomposed to an environmental standard value without any problem and 1,1-DCE could be decomposed.
「実験２」
１，１−ＤＣＥをジクロロメタン（ＤＣＭ）とした以外は、実験１と同様にして下記５つの条件を用意して行った。
条件６ ：Ｎ２３株及びＤＣＭを加えない（コントロール）
条件７ ：ＤＣＭを加えない（ポジティブコントロール）
条件８ ：ＤＣＭ濃度が１．０ｍｇ／Ｌ
条件９ ：ＤＣＭ濃度が５．０ｍｇ／Ｌ
条件１０：ＤＣＭ濃度が１０．０ｍｇ／Ｌ
培養開始１日後、４日後、７日後にサンプリングを行い、溶液中のＤＣＭ、１，４−ジオキサン濃度を測定した。
各条件の１，４−ジオキサン濃度及びＤＣＭ濃度の経時変化を図４、５に示す。なお、数値は２連の平均値を用いている。
"Experiment 2"
The following five conditions were prepared in the same manner as in Experiment 1 except that 1,1-DCE was changed to dichloromethane (DCM).
Condition 6: N23 strain and DCM are not added (control)
Condition 7: DCM is not added (positive control)
Condition 8: DCM concentration is 1.0 mg/L
Condition 9: DCM concentration is 5.0 mg/L
Condition 10: DCM concentration is 10.0 mg/L
Sampling was performed 1 day, 4 days, and 7 days after the start of culture, and the concentrations of DCM and 1,4-dioxane in the solution were measured.
Changes over time in the 1,4-dioxane concentration and the DCM concentration under each condition are shown in FIGS. In addition, the numerical value uses the average value of 2 consecutive.
Ｎ２３株が、ＤＣＭを生分解できることが確かめられた。
また、Ｎ２３株は、ＤＣＭの存在下でも、１，４−ジオキサンが分解できることが確かめられたが、ＤＣＭよりもジオキサンを先に分解することが確認できた。ＤＣＭ濃度が１０．０ｍｇ／Ｌ以下程度であれば、１，４−ジオキサンを問題なく環境基準値まで分解できることが確かめられたが、本条件では、ＤＣＭ濃度が５．０ｍｇ／Ｌ以上では、ＤＣＭを環境基準値まで分解できなかった。
It was confirmed that the N23 strain was able to biodegrade DCM.
It was also confirmed that the N23 strain can decompose 1,4-dioxane even in the presence of DCM, but it was confirmed that dioxane was decomposed earlier than DCM. It was confirmed that 1,4-dioxane can be decomposed to the environmental standard value without any problem if the DCM concentration is about 10.0 mg/L or less, but under these conditions, when the DCM concentration is 5.0 mg/L or more, DCM Could not be decomposed to the environmental standard value.
「実験３」
１２０ｍｌ容のバイアル瓶にＮ２３株（菌体濃度：１５０ｍｇ−ｄｒｙ ｃｅｌｌ／Ｌ）、シス−１，２−ＤＣＥ（１０ｍｇ／Ｌ）あるいはクロロエチレン（２ｍｇ／Ｌ）、無機塩培地を合計で４０ｍＬとなるように添加した。その後、ブチルゴム栓で密栓し３０℃、１２０ｒｐｍの条件で振盪しながら試験を行った（Ｎ＝２）。実験は菌を入れた条件と入れていない条件（コントロール）の２通り実施した。
培養開始０日後、１日後、２日後、３日後、４日後、７日後にサンプリングを行い、溶液中のシス−１，２−ＤＣＥあるいはクロロエチレン（ＶＣＭ）濃度を測定した。
１，４−ジオキサン濃度及びＤＣＭ濃度の経時変化を図６、７に示す。なお、数値は２連の平均値を用いている。
"Experiment 3"
A total of 40 mL of N23 strain (cell concentration: 150 mg-dry cell/L), cis-1,2-DCE (10 mg/L) or chloroethylene (2 mg/L), and an inorganic salt medium were added to a 120 ml vial bottle. Was added. Then, the test was carried out while sealing with a butyl rubber stopper and shaking under conditions of 30° C. and 120 rpm (N=2). The experiment was carried out in two ways, that is, the condition containing the bacteria and the condition not containing it (control).
Sampling was performed 0 day, 1 day, 2 days, 3 days, 4 days, and 7 days after the start of culture to measure the cis-1,2-DCE or chloroethylene (VCM) concentration in the solution.
The time-dependent changes in the 1,4-dioxane concentration and the DCM concentration are shown in FIGS. In addition, the numerical value uses the average value of 2 consecutive.
Ｎ２３株は、ジオキサンの非存在下で、シス−１，２−ＤＣＥ、ＶＣＭを分解できることが確認できた。すなわち、Ｎ２３株は、ジオキサン等を誘導物質として馴養することなく、有機塩素化合物を分解できることが確かめられた。 It was confirmed that the N23 strain can decompose cis-1,2-DCE and VCM in the absence of dioxane. That is, it was confirmed that the N23 strain can decompose organic chlorine compounds without acclimatizing dioxane as an inducer.
「実験４」
１２０ｍｌ容のバイアル瓶にＮ２３株（菌体濃度：５００ｍｇ−ｄｒｙ ｃｅｌｌ／Ｌ）、ＶＯＣ（１，１−ＤＣＥ、シス−１，２−ＤＣＥ、トリクロロエチレン、テトラクロロエチレン、ＤＣＭ、１，２−ジクロロエタン、１，１，１−トリクロロエタンの７物質を各１ｍｇ／Ｌ）、無機塩培地を合計で２４ｍＬとなるように添加した。その後、ブチルゴム栓で密栓し３０℃、１２０ｒｐｍの条件で振盪しながら試験を行った（Ｎ＝２）。
培養開始０日後、１日後、３日後、７日後にサンプリングを行い、溶液中のＶＯＣの各濃度を測定した。
各有機塩素化合物濃度の経時変化を図８、９に示す。なお、数値は２連の平均値を用いている。
"Experiment 4"
N23 strain (cell concentration: 500 mg-dry cell/L), VOC (1,1-DCE, cis-1,2-DCE, trichloroethylene, tetrachloroethylene, DCM, 1,2-dichloroethane, 1 , 1,1-trichloroethane (7 substances each 1 mg/L) and an inorganic salt medium were added so that the total amount was 24 mL. Then, the test was carried out while sealing with a butyl rubber stopper and shaking at 30° C. and 120 rpm (N=2).
Sampling was performed 0 day, 1 day, 3 days, and 7 days after the start of culture to measure each concentration of VOC in the solution.
8 and 9 show changes with time of the respective organic chlorine compound concentrations. In addition, the numerical value uses the average value of 2 consecutive.
Ｎ２３株が、複数種の有機塩素化合物を同時に分解できることが確かめられた。特に、炭素原子数１〜３、塩素原子数１〜２の有機塩素化合物である１，１−ＤＣＥ、シス−１，２−ＤＣＥ、ＤＣＭ、１，２−ジクロロエタンに対して優れた生分解性を示した。 It was confirmed that the N23 strain can simultaneously decompose multiple kinds of organochlorine compounds. Particularly, excellent biodegradability for 1,1-DCE, cis-1,2-DCE, DCM, and 1,2-dichloroethane, which are organic chlorine compounds having 1 to 3 carbon atoms and 1 to 2 chlorine atoms. showed that.
「実験５」
１２０ｍｌ容のバイアル瓶にＮ２３株（菌体濃度：１０００ｍｇ−ｄｒｙ ｃｅｌｌ／Ｌ）、１，４−ジオキサン（添加濃度１ｍｇ／Ｌ）、ＶＯＣ（クロロエチレン、１，１−ＤＣＥ、シス−１，２−ＤＣＥ、ＤＣＭ、１，２−ジクロロエタン、１，２−ジクロロプロパン、１，３−ジクロロプロペンの７物質を各１ｍｇ／Ｌ）、無機塩培地を条件毎に合計で３０ｍＬとなるように添加した。その後、ブチルゴム栓で密栓し３０℃、１２０ｒｐｍの条件で振盪しながら試験を行った（Ｎ＝２）。条件は、菌を添加しない系（ｃｏｎｔｒｏｌ）と菌を添加した系（１０００ｐｐｍ）を用意した。
培養開始０日後、４日後にサンプリングを行い、溶液中の１，４−ジオキサン及びＶＯＣの各濃度を測定した。
図１０に０日目に対する４日目の各物質の除去率を示す。なお、数値は２連の平均値を用いている。
"Experiment 5"
N120 strain (bacterial cell concentration: 1000 mg-dry cell/L), 1,4-dioxane (additional concentration 1 mg/L), VOC (chloroethylene, 1,1-DCE, cis-1,2) in a 120 ml vial bottle. -DCE, DCM, 1,2-dichloroethane, 1,2-dichloropropane, 1,3-dichloropropene (7 substances each 1 mg/L), and an inorganic salt medium were added so that the total amount was 30 mL for each condition. .. Then, the test was carried out while sealing with a butyl rubber stopper and shaking at 30° C. and 120 rpm (N=2). As conditions, a system containing no bacteria (control) and a system containing bacteria (1000 ppm) were prepared.
Sampling was carried out 0 days and 4 days after the start of culture to measure the respective concentrations of 1,4-dioxane and VOC in the solution.
FIG. 10 shows the removal rate of each substance on the 4th day with respect to the 0th day. In addition, the numerical value uses the average value of 2 consecutive.
Ｎ２３株が１，４−ジオキサンと複数種の有機塩素化合物を同時に分解できることが確かめられた。また、シス−１，２−ＤＣＥ、１，２−ジクロロプロパン以外の有機塩素化合物は環境基準値以下まで低減された。
ｃｏｎｔｒｏｌにおいて、０日目と４日目で大きく濃度が変化した物質が見られるが、これは０日目のサンプリングの際に、一部の物質では気相と液相の平衡移動が完了する前に採取した影響であると考えられる。特に、１，１−ＤＣＥは初期の添加濃度が高くなってしまったことも除去率が高い原因であると考えられる。また、１，３−ジクロロプロペンは加水分解される特徴を持っているため、ｃｏｎｔｒｏｌでも大きく除去されたと思われる。
It was confirmed that the N23 strain can simultaneously decompose 1,4-dioxane and plural kinds of organic chlorine compounds. In addition, organochlorine compounds other than cis-1,2-DCE and 1,2-dichloropropane were reduced to below the environmental standard value.
In the control, there are some substances whose concentration changed significantly on the 0th day and the 4th day, but this was observed at the time of sampling on the 0th day before the equilibrium transfer of gas phase and liquid phase was completed for some substances. It is thought that it is the effect collected in. In particular, it is considered that 1,1-DCE has a high removal rate because the initial addition concentration is high. Further, since 1,3-dichloropropene has a characteristic of being hydrolyzed, it is considered that it was largely removed by control.
「実験６」
１２０ｍｌ容のバイアル瓶にＮ２３株（菌体濃度：５００ｍｇ−ｄｒｙ ｃｅｌｌ／Ｌ）、ＴＣＥ（０．１ｍｇ／Ｌ、１ｍｇ／Ｌ、１０ｍｇ／Ｌの３条件）、無機塩培地を合計で５０ｍＬとなるように添加した。また、それぞれについてＮ２３株を含まないネガティブコントロールを調製した。その後、ブチルゴム栓で密栓し３０℃、１２０ｒｐｍの条件で振盪しながら試験を行った（Ｎ＝２）。
培養開始０日後、１日後、２日後、７日後、１４日後、２１日後にサンプリングを行い、溶液中のＴＣＥ濃度を測定した。
ＴＣＥ濃度の経時変化を図１１に示す。（Ａ）が初期ＴＣＥ濃度０．１ｍｇ／Ｌ、（Ｂ）が初期ＴＣＥ濃度１ｍｇ／Ｌ、（Ｃ）が初期ＴＣＥ濃度１０ｍｇ／Ｌである。なお、数値は２連の平均値を用いている。
"Experiment 6"
A total of 50 mL of N23 strain (cell concentration: 500 mg-dry cell/L), TCE (3 conditions of 0.1 mg/L, 1 mg/L, and 10 mg/L) and an inorganic salt medium are added to a 120 ml vial bottle. Was added as. In addition, a negative control containing no N23 strain was prepared for each. Then, the test was carried out while sealing with a butyl rubber stopper and shaking at 30° C. and 120 rpm (N=2).
Sampling was performed 0 day, 1 day, 2 days, 7 days, 14 days, and 21 days after the start of culture to measure the TCE concentration in the solution.
The time-dependent change in TCE concentration is shown in FIG. (A) has an initial TCE concentration of 0.1 mg/L, (B) has an initial TCE concentration of 1 mg/L, and (C) has an initial TCE concentration of 10 mg/L. In addition, the numerical value uses the average value of 2 consecutive.
Ｎ２３株は、分解速度は遅いものの、ジオキサンの非存在下で、ＴＣＥを分解できることが確認できた。すなわち、Ｎ２３株は、ジオキサン等を誘導物質として馴養することなく、ＴＣＥを分解できることが確かめられた。 It was confirmed that the N23 strain can decompose TCE in the absence of dioxane, although the decomposition rate is slow. That is, it was confirmed that the N23 strain can decompose TCE without acclimatizing dioxane as an inducer.
「実験７」
１２０ｍｌ容のバイアル瓶にＮ２３株（菌体濃度：５００ｍｇ−ｄｒｙ ｃｅｌｌ／Ｌ）、１，４−ジオキサン（１０ｍｇ／Ｌ）、ＴＣＥ（０．１ｍｇ／Ｌ、１ｍｇ／Ｌ、１０ｍｇ／Ｌの３条件）、無機塩培地を合計で５０ｍＬとなるように添加した。その後、ブチルゴム栓で密栓し３０℃、１２０ｒｐｍの条件で振盪しながら試験を行った。
実験は下記５つの条件を用意して行った。
条件１１：Ｎ２３株及びジオキサンを加えない（コントロール）
条件１２：ＴＣＥを加えない（ポジティブコントロール）
条件１３：ＴＣＥ濃度が０．１ｍｇ／Ｌ、ジオキサン濃度が１０ｍｇ／Ｌ
条件１４：ＴＣＥ濃度が１ｍｇ／Ｌ、ジオキサン濃度が１０ｍｇ／Ｌ
条件１５：ＴＣＥ濃度が１０ｍｇ／Ｌ、ジオキサン濃度が１０ｍｇ／Ｌ
"Experiment 7"
3 conditions of N23 strain (cell concentration: 500 mg-dry cell/L), 1,4-dioxane (10 mg/L), TCE (0.1 mg/L, 1 mg/L, 10 mg/L) in a 120 ml capacity vial. ), and an inorganic salt medium was added so that the total amount was 50 mL. Then, the test was carried out by sealing with a butyl rubber stopper and shaking at 30° C. and 120 rpm.
The experiment was conducted under the following five conditions.
Condition 11: N23 strain and dioxane are not added (control)
Condition 12: TCE is not added (positive control)
Condition 13: TCE concentration is 0.1 mg/L, dioxane concentration is 10 mg/L
Condition 14: TCE concentration is 1 mg/L, dioxane concentration is 10 mg/L
Condition 15: TCE concentration 10 mg/L, dioxane concentration 10 mg/L
培養開始１日後、２日後、５日後、７日後にサンプリングを行い、溶液中のＴＣＥ、１，４−ジオキサン濃度を測定した。
各条件の１，４−ジオキサン濃度及びＴＣＥ濃度の経時変化を、それぞれ図１２、１３に示す。なお、数値は２連の平均値を用いている。
Sampling was performed 1 day, 2 days, 5 days, and 7 days after the start of culture to measure the TCE and 1,4-dioxane concentrations in the solution.
The time-dependent changes in the 1,4-dioxane concentration and the TCE concentration under each condition are shown in FIGS. In addition, the numerical value uses the average value of 2 consecutive.
Ｎ２３株が、ジオキサン存在下でも、ＴＣＥを分解できることが確認できた。また、Ｎ２３株が、ＴＣＥとジオキサンとを同時に分解できることが確認できた。 It was confirmed that the N23 strain can decompose TCE even in the presence of dioxane. Further, it was confirmed that the N23 strain can simultaneously decompose TCE and dioxane.
「実験８」
１２０ｍｌ容のバイアル瓶にＮ２３株（菌体濃度：５００ｍｇ−ｄｒｙ ｃｅｌｌ／Ｌ）、１，１，２−ＴＣＡ（０．１ｍｇ／Ｌ、１ｍｇ／Ｌ、１０ｍｇ／Ｌの３条件）、無機塩培地を合計で５０ｍＬとなるように添加した。また、それぞれについてＮ２３株を含まないネガティブコントロールを調製した。その後、ブチルゴム栓で密栓し３０℃、１２０ｒｐｍの条件で振盪しながら試験を行った（Ｎ＝２）。
培養開始５日後にサンプリングを行い、溶液中の１，１，２−ＴＣＡ濃度を測定した。
ＴＣＥ濃度の経時変化を図１４に示す。なお、数値は２連の平均値を用いている。
"Experiment 8"
N23 strain (cell concentration: 500 mg-dry cell/L), 1,1,2-TCA (0.1 mg/L, 1 mg/L, 3 conditions of 10 mg/L), inorganic salt medium in a 120 ml vial. Was added to a total of 50 mL. In addition, a negative control containing no N23 strain was prepared for each. Then, the test was carried out while sealing with a butyl rubber stopper and shaking at 30° C. and 120 rpm (N=2).
Sampling was performed 5 days after the start of culturing, and the 1,1,2-TCA concentration in the solution was measured.
FIG. 14 shows the change over time in the TCE concentration. In addition, the numerical value uses the average value of 2 consecutive.
ネガティブコントロールでは、１，１，２−ＴＣＡ濃度に変化はなかった（図１４Ａ）。一方、Ｎ２３株を加えた系では、初期１，１，２−ＴＣＡ添加濃度０．１ｍｇ／Ｌ、１ｍｇ／Ｌでは、５日目までに検出下限値以下まで減少し、初期１，１，２−ＴＣＡ濃度１０ｍｇ／Ｌの系でも、減少傾向が確認できた（図１４Ｂ）。 In the negative control, there was no change in the 1,1,2-TCA concentration (Fig. 14A). On the other hand, in the system in which the N23 strain was added, the initial 1,1,2-TCA addition concentrations of 0.1 mg/L and 1 mg/L decreased to below the detection lower limit value by the 5th day, and the initial 1,1,2- A decrease tendency was confirmed even in the system with a TCA concentration of 10 mg/L (FIG. 14B).
「実験９」
１２０ｍｌ容のバイアル瓶にＮ２３株（菌体濃度：５００ｍｇ−ｄｒｙ ｃｅｌｌ／Ｌ）、１，４−ジオキサン（１０ｍｇ／Ｌ）、１，１，２−ＴＣＥ（０．１ｍｇ／Ｌ、１ｍｇ／Ｌ、１０ｍｇ／Ｌの３条件）、無機塩培地を合計で５０ｍＬとなるように添加した。その後、ブチルゴム栓で密栓し３０℃、１２０ｒｐｍの条件で振盪しながら試験を行った。条件は、１）Ｎ２３株無し（ネガティブコントロール）、２）Ｎ２３株ありの２条件を作成した。各条件、２連で試験を行い、１，４−ジオキサン、１，１，２−ＴＣＡ濃度を測定した。
"Experiment 9"
N120 strain (cell concentration: 500 mg-dry cell/L), 1,4-dioxane (10 mg/L), 1,1,2-TCE (0.1 mg/L, 1 mg/L) in a 120 ml vial bottle. (3 conditions of 10 mg/L) and an inorganic salt medium were added so that the total amount was 50 mL. Then, the test was carried out by sealing with a butyl rubber stopper and shaking at 30° C. and 120 rpm. Two conditions were prepared: 1) without N23 strain (negative control) and 2) with N23 strain. The test was conducted in duplicate under each condition, and the 1,4-dioxane and 1,1,2-TCA concentrations were measured.
培養開始１日後、２日後、５日後、７日後にサンプリングを行い、溶液中の１，４−ジオキサン、１，１，２−ＴＣＡの濃度を測定した。
各条件の１，４−ジオキサン濃度及び１，１，２−ＴＣＡ濃度の経時変化を、それぞれ図１５、１６に示す。なお、数値は２連の平均値を用いている。
Sampling was performed 1 day, 2 days, 5 days, and 7 days after the start of culturing to measure the concentrations of 1,4-dioxane and 1,1,2-TCA in the solution.
The time-dependent changes in the 1,4-dioxane concentration and the 1,1,2-TCA concentration under each condition are shown in FIGS. In addition, the numerical value uses the average value of 2 consecutive.
ネガティブコントロールではいずれの物質も濃度変化はなかった（図１５Ａ、図１６Ａ）。
Ｎ２３株を添加した系は、１，１，２−ＴＣＡの初期濃度０．１ｍｇ／Ｌ、１ｍｇ／Ｌの条件では、１，４−ジオキサン、１，１，２−ＴＣＡともに３日目までに検出下限値まで減少した。１，１，２−ＴＣＡの初期濃度１０ｍｇ／Ｌの条件では、１，４−ジオキサンは７日目までに検出下限値まで減少した。一方、１，１，２−ＴＣＡは７日目時点においても高い濃度で残存したが、ネガティブコントロールと比較して減少傾向が確認できた（図１５Ｂ、図１６Ｂ）。
There was no change in the concentration of any substance in the negative control (Fig. 15A, Fig. 16A).
In the system to which the N23 strain was added, both 1,4-dioxane and 1,1,2-TCA were prepared by the 3rd day under the conditions of initial concentrations of 1,1,2-TCA of 0.1 mg/L and 1 mg/L. It decreased to the lower limit of detection. Under the condition that the initial concentration of 1,1,2-TCA was 10 mg/L, 1,4-dioxane decreased to the lower limit of detection by day 7. On the other hand, 1,1,2-TCA remained at a high concentration even on the 7th day, but a decreasing tendency could be confirmed as compared with the negative control (FIGS. 15B and 16B ).
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JPH09276841A (en) * | 1996-04-12 | 1997-10-28 | Canon Inc | Method and apparatus for purifying contaminated soil |
| WO2016181802A1 (en) * | 2015-05-11 | 2016-11-17 | 大成建設株式会社 | Constitutive 1,4-dioxane-degrading bacterium |
| JP2018094455A (en) * | 2016-12-08 | 2018-06-21 | 大成建設株式会社 | Method for biological decomposition treatment of cyclic ether |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH09276841A (en) * | 1996-04-12 | 1997-10-28 | Canon Inc | Method and apparatus for purifying contaminated soil |
| WO2016181802A1 (en) * | 2015-05-11 | 2016-11-17 | 大成建設株式会社 | Constitutive 1,4-dioxane-degrading bacterium |
| JP2018094455A (en) * | 2016-12-08 | 2018-06-21 | 大成建設株式会社 | Method for biological decomposition treatment of cyclic ether |
Non-Patent Citations (4)
| Title |
|---|
| CHEMOSPHERE, vol. 91, JPN6022039203, 2013, pages 88 - 92, ISSN: 0004876712 * |
| ENVIRONMENTAL SCIENCE & TECHNOLOGY LETTERS, vol. 6, JPN6022049236, December 2018 (2018-12-01), pages 49 - 54, ISSN: 0004925437 * |
| ENVIRONMENTAL SCIENCE & TECHNOLOGY, vol. 50, JPN6022049237, August 2016 (2016-08-01), pages 9599 - 9607, ISSN: 0004925436 * |
| 生物工学会誌, vol. 第95巻, 第9号, JPN7022004454, 2017, pages 547, ISSN: 0004876711 * |
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