JPH0824905B2 - Purification method of contaminated soil using aerobic microorganisms - Google Patents

Purification method of contaminated soil using aerobic microorganisms


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JPH0824905B2 JP2023593A JP2023593A JPH0824905B2 JP H0824905 B2 JPH0824905 B2 JP H0824905B2 JP 2023593 A JP2023593 A JP 2023593A JP 2023593 A JP2023593 A JP 2023593A JP H0824905 B2 JPH0824905 B2 JP H0824905B2
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contaminated soil
purification method
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JPH06226230A (en )
裕夫 内山
修身 矢木
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【0001】 [0001]

【産業上の利用分野】本発明は、特定の微生物を用いて、汚染土壌、とくに有機塩素化合物によって汚染された土壌の浄化法に関する。 The present invention relates to using a specific microorganism, contaminated soil, particularly to the purification method of soil contaminated by chlorinated organic compounds.

【0002】 [0002]

【従来の技術】工場からの排水又は排ガス、或いは土壌中には各種有機塩素化合物が混入されており、近時、環境汚染等の問題から、これらの有効な除去が注目されるところとなっている。 Wastewater or exhaust gas of the Related Art Plant, or various organic chlorine compounds have been mixed in the soil, recent, from such environmental pollution problem, a place where these effective removal is noted there.

【0003】殊にトリクロロエチレン(TCE)は、I [0003] In particular trichlorethylene (TCE) is, I
C産業等で用いられている難分解性化合物であり、発ガン性を有し、地下水あるいは土壌汚染物質として問題になっている。 A hardly decomposable compound used in the C industries have carcinogenic, at issue as groundwater or soil contamination.

【0004】従来、排水中或いは排ガス中から、トリクロロエチレンのような有機塩素化合物を除去する方法としては、活性炭による吸着除去法や微生物を用いる方法もいくつか試みられ報告されている。 Conventionally, from or during the exhaust gas effluent, as a method for removing organic chlorine compounds such as trichlorethylene, a method of using adsorption removal method or a microorganism with activated carbon it has been reported several attempted.

【0005】例えば、ロドトルラ属、クラドスポリウム属、キャンデイダ属、サッカロミセス属及びストレプトミセス属の微生物等を用いてポリクロル化されたビフェニルのような有機塩素化合物を分解除去する例(特開昭48−98085号、特開昭48−98086号、特開昭49−6186号)、及びメチロシナス属、メチロシスチス属、メチロコッカス属及びメチロバクテリウム属の細菌のようなメタン資化性細菌を用いて、m−クロルトルエンのようなハロゲン置換ベンゼンを分解する例(特開昭55−127196号)が報告されている。 [0005] For example, Rhodotorula, Cladosporium spp, Kyandeida genus example decomposition to remove organic chlorine compounds such as Porikuroru of biphenyl by using the microorganisms of the Saccharomyces genus and Streptomyces (JP 48- No. 98085, JP-48-98086, JP-49-6186), and Mechiroshinasu genus Mechiroshisuchisu genus using methane-assimilating bacteria such as Methylococcus genera and Methylobacterium genus bacteria, m - example decompose halogen-substituted benzene such as chlorotoluene (JP 55-127196) it has been reported.

【0006】しかしながらトリクロロエチレン及びその類縁化合物のような脂肪族塩素化合物を有効に分解除去する微生物についてはほとんど報告されておらず、わずかに本出願人による提案(特開平2−92274号)があるのみである。 However it not has been little reported for microorganisms to effectively decompose and remove the aliphatic chlorinated compounds such as trichlorethylene and its related compounds, only a slight proposed by the present applicant (Japanese Patent Laid-Open No. 2-92274) it is.

【0007】 [0007]

【発明が解決しようとする課題】ところで、全国の多くの地下水がトリクロロエチレンのような、揮発性有機塩素化合物によって汚染されていることが見出されており、汚染地下水の浄化法が検討されているが、汚染地下水を浄化するためには汚染土壌を浄化することが根本的に解決するために必須となる。 [SUMMARY OF THE INVENTION Incidentally, like many groundwater trichlorethylene nationwide have been found to be contaminated by the volatile organic chlorine compounds, the purification method of the polluted groundwater has been studied but, in order to purify the contaminated groundwater to purify the contaminated soil is essential in order to solve fundamentally.

【0008】従来、土壌中からトリクロロエチレンのような有機塩素化合物を除去するには、土壌を掘削し風乾する方法あるいは真空吸引装置を用いて土壌中のガスを吸引する方法が用いられてきたが、掘削には多大の費用を要し、また吸引除去は低濃度の除去が困難等の問題点がある。 Conventionally, in removing the organic chlorine compounds such as trichlorethylene in the soil, a method of sucking the gas in the soil using a method or vacuum suction device air dried excavated soil have been used, the drilling takes great expense, also aspirated low density of removal there is a problem of difficulty or the like.

【0009】また、風乾、および吸引除去のいずれの場合も、系外に除去するもので、分解し無害化するものではない。 Further, air-dried, and the suction in either case of the removal, but to remove from the system, does not decompose and detoxify.

【0010】一方、土壌中の有機塩素化合物の効率的かつ簡便な分解除去手段として、好気性微生物を用いて土壌中のトリクロロエチレン及びその類塩化合物のような脂肪族炭化水素を有効に分解除去する方法については報告されていない。 On the other hand, as an efficient and simple decomposition removal means of the organic chlorine compounds in the soil, effectively decompose removing aliphatic hydrocarbons such as trichlorethylene and its class salt compounds in soil using aerobic microorganisms It has not been reported how.

【0011】本発明は、こうした実情の下に、有機塩素化合物、とくに脂肪族塩素化合物により汚染された土壌を微生物を利用して簡便な方法により有効に浄化する方法を提供することを目的とするものである。 [0011] The present invention is, under these circumstances, organic chlorine compounds, and to provide a method for effectively purifying the particularly simple method and contaminated soil using microorganisms by aliphatic chlorine compounds it is intended.

【0012】 [0012]

【課題を解決するための手段】本発明者は、鋭意検討した結果、特定の好気性菌を易酸素発生性物質とともに汚染土壌に散布することにより解決し得ることを知見し、 The present inventors SUMMARY OF THE INVENTION a result of extensive studies, and found that may be solved by spraying the contaminated soil certain aerobic bacteria with ease oxygen generating material,
本発明を完成するに至った。 This has led to the completion of the present invention.

【0013】すなわち、メチロシナス(Methylo [0013] In other words, Mechiroshinasu (Methylo
sinus)属に属し、脂肪族塩素化合物分解能を有する微生物を、易酸素発生性物質とともに土壌に散布する汚染土壌の浄化法を要旨とするものである。 sinus) belonging to the genus microorganisms having an aliphatic chlorine compounds resolution, it is an purifying method gist of contaminated soil to spray the soil with ease oxygen generating material.

【0014】本発明に使用する微生物は、メチロシナス(Methylosinus)属に属し、脂肪族塩素化合物分解能を有する微生物がある。 [0014] Microorganisms used in the present invention belongs to Mechiroshinasu (Methylosinus) genus, there is a microorganism having an aliphatic chlorine compounds resolution. この微生物自体は、 The organism itself,
特開平2−92274号公報に開示されている。 It disclosed in Japanese Patent Laid-Open No. 2-92274.

【0015】これは各種土壌に広く分布しこれから採取し得られるが、その採取の方法としては、例えば次のような方法を用いる。 [0015] This is obtained widely distributed now taken various soils, as a method of collecting, using for example, the following method.

【0016】すなわち、培養はブチルゴム栓及びアルミシールで密閉したバイアル瓶を用い、30℃にて振とうする。 [0016] That is, the culture using a vial closed with a butyl rubber stopper and aluminum seal, shaken at 30 ° C.. トリクロロエチレン量はヘッドスペースより気相を一定量取り、ガスクロマトグラフィーにより定量し、 Trichlorethylene amount takes a certain amount of the gas phase from the head space was quantified by gas chromatography,
ヘンリーの法則式より液相濃度を算出する。 To calculate the liquid phase concentration than the law expressions of Henry.

【0017】前記手段を用い、例えば採取した土壌を1 [0017] using the means, for example, collected soil 1
ppmトリクロロエチレン及びメタンの存在下で集積培養を繰り返し、トリクロロエチレンをよく分解する混合微生物系を得る。 ppm repeated enrichment culture in the presence of trichlorethylene and methane, obtained with decomposing mixed microbial systems and trichlorethylene. トリクロロエチレンの分解には酸素及びメタンが必須であることから、混合微生物系からメタノトローフの単離を行う。 Since the degradation of trichlorethylene oxygen and methane is required, it performs the isolation of methanotrophic from mixed microbial system.

【0018】本発明において単離された菌は、公知のメチロシナス・トリコスポリウムに属するメチロシナス・ [0018] The isolated bacteria in the present invention, Mechiroshinasu, belonging to the well-known Mechiroshinasu-Trico sports helium
トリコスポリウム・TSUKUBAである。 Is a Trichoderma spot potassium · TSUKUBA.

【0019】この菌を顕微鏡で観察すると、巾0.6〜 [0019] When observing the bacteria under a microscope, width 0.6
1μm、長さ1〜5μmの短桿菌で以下の表に示すような特性を有するものである。 1 [mu] m, and has a characteristic as shown in the table below in short bacillus length 1 to 5 [mu] m.

【0020】 Chracteristics of methane−utilizing bacterium Gram stain Negative Cell shape Short rod Number of flagela 0 Motility − Growth on methane + ethane − propane − n−butane − dimethylether − methylamine − methanol + ethanol − nutrient broth − Growth at 30℃ + 37℃ + 45℃ − Mol% G+C DNA 64.5 Major fatty acid C 18:1 (96.5%) Hydroxy fatty acid type 2−0H Quinone type Q 8以上の菌学的性質に基づき、本 [0020] Chracteristics of methane-utilizing bacterium Gram stain Negative Cell shape Short rod Number of flagela 0 Motility - Growth on methane + ethane - propane - n-butane - dimethylether - methylamine - methanol + ethanol - nutrient broth - Growth at 30 ℃ + 37 ℃ + 45 ℃ - Mol% G + C DNA 64.5 Major fatty acid C 18: 1 based on (96.5%) Hydroxy fatty acid type 2-0H quinone type Q 8 or more microbiological properties, this 発明のメチロシナス菌株の同定を行った。 The identification of Mechiroshinasu strains of the invention were carried out.

【0021】本発明のメチロシナスの菌株は、菌の形態、グラム染色などの顕微鏡的所見、生理学的諸性質などから、公知菌メチロシナス・トリコスポリウムOB [0021] Strains of Mechiroshinasu of the present invention, the form of bacteria, microscopic findings, such as Gram staining, and the like physiological properties, known bacteria Mechiroshinasu & Trichoderma sports potassium OB
3bの性状について記載した文献{1. 3b documents describing properties of {1. Journal Journal
of General Microbiology of General Microbiology
61 ,205−218(1970)、2. 61, 205-218 (1970), 2. Microb Microb
ial Growth on C 1 Compound ial Growth on C 1 Compound
s p. s p. 123〜133(1984)、3. 123-133 (1984), 3. Journ Journ
al of General AppliedMicr al of General AppliedMicr
obiology 33,135〜165(198 obiology 33,135~165 (198
7)}に記されているWhittenburyら、及び駒形らの分類に基づき、メチロシナス・トリコスポリウムOB 3bに近縁の株と同定された。 7) Whittenbury et al., Which is described in}, and on the basis of Komagata et al classification, it was identified as strains closely related to Mechiroshinasu-Trico sports potassium OB 3b.

【0022】しかしながら、鞭毛を有せずC 19の飽和脂肪酸も有せず、又、ロゼットを形成しない点で、メチロシナス・トリコスポリウムOB 3bとは明らかに相違し、新菌株と同定され、メチロシナス・トリコスポリウム・TSUKUBAと命名された。 [0022] However, saturated fatty acids C 19 does not have a flagella also not closed, and in that it does not form rosettes, clearly different from the Mechiroshinasu-Trico sports potassium OB 3b, identified as novel strain, Mechiroshinasu - it was named Trico spot potassium · TSUKUBA.

【0023】本発明の菌は工業技術院微生物工業技術研究所に微工研菌寄第10004号として寄託されている。 [0023] The bacteria of the present invention has been deposited as Biko Institute bacteria nearest No. 10004 in the Fermentation Research Institute Agency of Industrial Science and Technology.

【0024】本発明の菌はトリクロロエチレン及びその各種類縁化合物、すなわち、シス−1,2−ジクロロエチレン、トランス−1,2−ジクロロエチレン、1,1 [0024] bacteria of the present invention is trichlorethylene and its various related compounds, i.e., cis-1,2-dichloroethylene, trans-1,2-dichloroethylene, 1,1
−ジクロロエチレン、1,1,2,2,−テトラクロロエタン、1,1,2−トリクロロエタン、1,2−ジクロロエタン、クロロホルムを分解する性質を有し、10 - dichloroethylene, 1,1,2,2 - tetrachloroethane, 1,1,2-trichloroethane, 1,2-dichloroethane, has a property of decomposing chloroform, 10
ppmの高濃度トリクロロエチレンを10日間で約半分に分解する能力を持つ。 The ppm high concentration of trichlorethylene with decomposing ability to about half at 10 days.

【0025】本発明浄化法を実施するにあたっては、微生物を予め適当な培地にて培養し、それを有機塩素化合物で汚染された土壌に散布する。 [0025] In practicing the present invention purification method, the microorganisms are cultured in advance a suitable medium, spraying the contaminated soil it with organic chlorine compounds. このとき、前記好気性微生物の分解活性を高めるため易酸素発生性物質を併用することが重要である。 At this time, it is important to use a readily oxygen generating material to enhance the degradation activity of the aerobic microorganisms. この併用により汚染土壌内を容易に好気性に保持することができ、効果的に土壌の浄化を行うことができる。 This combination makes it possible to hold the easily aerobic within contaminated soil, effectively it can be purified soil.

【0026】前記易酸素発生性物質としては、土壌中において容易に酸素を発生することができ、しかも土壌を新たに汚染するような有害な残留品を生じることのないものであればとくに制限はない。 [0026] Examples of the easy-oxygen generating material, it is possible to generate easily the oxygen in the soil, yet particularly limited as long as not to cause harmful residual products such as newly contaminate the soil Absent. このような易酸素発生性物質として、とくに過酸化水素、過酸化カルシウム、 Such as a free oxygen-generating substance, particularly hydrogen peroxide, calcium peroxide,
過酸化マグネシウムなどが好ましく、最も好ましいものは過酸化水素である。 Is preferable, magnesium peroxide, and most preferred is hydrogen peroxide.

【0027】微生物及び易酸素発生性物質の土壌への散布法は、培養液及び易酸素発生性物質それ自体を直接散布しても、又、蒸留水などで希釈して散布してもよい。 The application method to the soil microorganisms and easy oxygen generating materials can be sprayed with broth and easy oxygen generating material itself directly, or may be sprayed diluted with distilled water etc..

【0028】 [0028]

【実施例】以下に実施例を示し、本発明をさらに詳細に説明する。 EXAMPLES the following examples are further illustrative of this invention.

【0029】(菌体の調製法) [0029] (Preparation of bacterial cells)

【0030】 [0030]

【表1】 [Table 1]

【0031】2リットルの培養装置に表1に示す培地を1リットル加え、これにメチロシナス・トリコスポリウム・TSUKUBA保存株10mgを接種する。 [0031] The medium shown in culture apparatus 2 liters in Table 1 was added 1 liter to which is inoculated with Mechiroshinasu-Trichoderma spot Agrobacterium-TSUKUBA stocks 10 mg. メタンガス濃度16v/v%の空気、メタンガス混合ガスを1 Methane concentration 16v / v% of air, the methane gas mixture 1
分間に1リットルの速度で連続的に通気し、30℃で、 Continuously aerated with 1 liter of speed minutes, at 30 ° C.,
48時間撹拌培養を行う。 Perform 48 hours of stirring culture. 菌体は培養後遠心分離により集菌し、−70℃で保存する。 Cells were harvested by centrifugation after cultivation, and stored at -70 ° C..

【0032】実施例 0.1ppmのトリクロロエチレンで汚染した土壌に対し、上記のようにして培養した培養液を、土壌1g当り菌10 6個となるように散布した。 [0032] For soil contaminated with trichlorethylene Example 0.1 ppm, the culture broth cultured as described above, were sprayed such that the bacteria 10 6 per soil 1g. 同時に過酸化水素を5ppmおよび10ppmとなるよう散布した。 At the same time it was sprayed so that hydrogen peroxide and 5ppm and 10 ppm.

【0033】その結果を比較例とともに以下に示す。 [0033] shown below together with comparative examples and the results.

【0034】 [0034]

【表2】 [Table 2]

【0035】 [0035]

【発明の効果】以上説明したように、本発明によれば有機塩素化合物で汚染された土壌を簡便な方法により効果的に浄化することができる。 As described above, according to the present invention, a soil contaminated with organic chlorine compounds can be efficiently purified by a simple method according to the present invention.

───────────────────────────────────────────────────── フロントページの続き (51)Int.Cl. 6識別記号 庁内整理番号 FI 技術表示箇所 // C02F 11/02 ZAB (C12N 1/20 C12R 1:01) ────────────────────────────────────────────────── ─── front page of continued (51) Int.Cl. 6 in identification symbol Agency Docket No. FI art display portion // C02F 11/02 ZAB (C12N 1/20 C12R 1:01)

Claims (4)

    【特許請求の範囲】 [The claims]
  1. 【請求項1】 メチロシナス(Methylosinu [Claim 1] Mechiroshinasu (Methylosinu
    s)属に属し、脂肪族塩素化合物分解能を有する微生物を、易酸素発生性物質とともに土壌に散布することを特徴とする汚染土壌の浄化法。 s) belonging to the genus purification method of contaminated soil, which comprises spraying a microorganism having an aliphatic chlorine compounds resolution, the soil with ease oxygen generating material.
  2. 【請求項2】 微生物がトリクロロエチレンを分解するメタン資化性細菌である請求項1記載の汚染土壌の浄化法。 2. A purification method of contaminated soil according to claim 1, wherein the microorganism is methane-utilizing bacteria to degrade trichlorethylene.
  3. 【請求項3】 微生物がメチロシナス・トリコスポリウム・TSUKUBA(微工研菌寄No.10004)である請求項1又は2記載の汚染土壌の浄化法。 3. A cleaning method of contaminated soil according to claim 1 or 2 wherein the microorganism is Mechiroshinasu & Trichoderma spot potassium-TSUKUBA (BikokenkinYadoriki No.10004).
  4. 【請求項4】 易酸素発生性物質が過酸化物である請求項1〜3に記載の汚染土壌の浄化法。 Purification method of contaminated soil according to claim 1 wherein the easily oxygen generating material is a peroxide.
JP2023593A 1993-02-08 1993-02-08 Purification method of contaminated soil using aerobic microorganisms Expired - Lifetime JPH0824905B2 (en)

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