JP3094072B2 - Novel microorganism and method for treating polycyclic aromatic compound-containing wastewater using the microorganism - Google Patents
Novel microorganism and method for treating polycyclic aromatic compound-containing wastewater using the microorganismInfo
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- JP3094072B2 JP3094072B2 JP983999A JP983999A JP3094072B2 JP 3094072 B2 JP3094072 B2 JP 3094072B2 JP 983999 A JP983999 A JP 983999A JP 983999 A JP983999 A JP 983999A JP 3094072 B2 JP3094072 B2 JP 3094072B2
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- polycyclic aromatic
- aromatic compound
- microorganism
- phenol
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Description
【０００１】[0001]
【発明の属する技術分野】本発明は、新規微生物、該微
生物に耐フェノール性を付与する方法及び該微生物を用
いる多環芳香族化合物含有排水の処理方法に関する。TECHNICAL FIELD The present invention relates to a novel microorganism, a method for imparting phenol resistance to the microorganism, and a method for treating polycyclic aromatic compound-containing wastewater using the microorganism.
【０００２】[0002]
【従来の技術】多環芳香族化合物は、生物難分解性化合
物であり、ＢＯＤ、ＣＯＤ等による排水規制では規制す
ることが難しく、これらの多環芳香族化合物は、活性汚
泥処理施設等の排水処理施設で分解されないまま環境中
に排出されていると思われる。これらの化合物が排出さ
れると、河川流域や河川の流れ込む湾内には、生物難分
解性の環境汚染物質として蓄積されので、この防止対策
が必要となる。又、近年、石油留出事故等により石油留
分中に含まれるこれらの化合物による海洋汚染が問題と
なっており、微生物による環境修復が注目されている。2. Description of the Related Art Polycyclic aromatic compounds are hardly biodegradable compounds, and are difficult to regulate by wastewater regulations such as BOD and COD. It is considered that they are released into the environment without being decomposed in the treatment facility. When these compounds are released, they are accumulated as biodegradable environmental pollutants in river basins and in bays into which rivers flow, so it is necessary to take measures to prevent them. In recent years, marine pollution by these compounds contained in petroleum fractions due to petroleum distilling accidents and the like has become a problem, and attention has been paid to environmental restoration by microorganisms.
【０００３】[0003]
【発明が解決しようとする課題】従って、本発明の目的
は、生物難分解性の多環芳香族化合物を分解する微生物
及び多環芳香族化合物を含有する排水の処理方法を提供
することである。SUMMARY OF THE INVENTION Accordingly, an object of the present invention is to provide a microorganism which decomposes a polycyclic aromatic compound which is hardly biodegradable and a method for treating wastewater containing the polycyclic aromatic compound. .
【０００４】[0004]
【課題を解決するための手段】上記の目的は以下の発明
によって達成される。即ち、本発明は、複素環を有する
多環芳香族化合物を分解するPseudomonas paucimobilis
菌及びRhodococcus ruber菌（以下、「新規微生物」或
いは単に「微生物」等と称することもある）、これらに
耐フェノール性を付与する方法及びこれらの微生物を用
いる多環芳香族化合物含有排水の処理方法である。The above object is achieved by the following invention. That is, the present invention relates to Pseudomonas paucimobilis which decomposes a polycyclic aromatic compound having a heterocyclic ring.
Bacteria and Rhodococcus ruber bacteria (hereinafter referred to as "new microorganisms"
Or simply referred to as "microorganisms") , a method for imparting phenol resistance thereto, and a method for treating polycyclic aromatic compound-containing wastewater using these microorganisms.
【０００５】生物難分解性多環芳香族化合物含有排水の
処理において、新規な微生物を用いることによって、排
水中の多環芳香族化合物を有効に除去することができ
る。又、これらの微生物は、活性汚泥に定着して、活性
汚泥の優先種となるので、本発明の微生物及び処理方法
は特に活性汚泥方式による排水処理に有用である。更
に、本発明の新規な微生物は、これに紫外線等により突
然変異を起こさせ、変異株をフェノール添加培地で馴養
することにより本発明の新規な微生物に耐フェノール性
が付与される。[0005] In the treatment of wastewater containing a biodegradable polycyclic aromatic compound, a novel microorganism can be used to effectively remove the polycyclic aromatic compound from the wastewater. In addition, since these microorganisms settle on activated sludge and become a priority species of activated sludge, the microorganism and the treatment method of the present invention are particularly useful for wastewater treatment by activated sludge. Furthermore, the novel microorganism of the present invention is mutated by ultraviolet rays or the like, and the mutant is acclimated in a phenol-containing medium to thereby impart phenol resistance to the novel microorganism of the present invention.
【０００６】[0006]
【発明の実施の形態】次に発明の実施の形態を挙げて本
発明を更に詳しく説明する。本発明の新規微生物は、多
環芳香族化合物を分解するPseudomonas paucimobilis
（シュウドモナス ポウシモビルス）菌及びRhodococcu
s ruber（ロードコッカス ルバー）菌であり、具体的
には、Pseudomonas paucimobilis ４２１Ｙ株（ＦＥＲ
Ｍ ＢＰ−５１２２）（以下ではＫＦ−１株と称す
る。）及びRhodococcus ruber ＴＡ ０９０２株（Ｆ
ＥＲＭ ＢＰ−５１２１）（以下ではＫＦ−２株と称す
る。）である。BEST MODE FOR CARRYING OUT THE INVENTION Next, the present invention will be described in more detail with reference to embodiments of the present invention. Novel microorganism of the present invention, a multi
Pseudomonas paucimobilis which degrades aromatic compounds
(Pseudomonas poshimovirus) and Rhodococcu
s ruber ( Rhodococcus ruber) , specifically, Pseudomonas paucimobilis 421Y strain (FER
MBP-5122) (hereinafter referred to as strain KF-1).
You. ) And Rhodococcus ruber TA0902 strain (F
ERM BP-5121) (hereinafter referred to as KF-2 strain)
You. ) .
【０００７】上記微生物は、１２０種の土壌から検出し
た多数の微生物を、７日間を１周期として３次まで集積
培養を行い、３次まで集積し、増殖が認められた微生物
については、該微生物を寒天培地上に塗布し、形成され
たコロニーより分離して得たものである。得られた微生
物のの菌学的性質を調べ、新規微生物であることを確認
した。得られた菌学的性質を下記の表に示す。[0007] The above microorganisms are obtained by culturing a large number of microorganisms detected from 120 kinds of soil up to the third order in one cycle of 7 days, and accumulating up to the third order. Was spread on an agar medium and separated from the formed colonies. The microbiological properties of the obtained microorganism were examined, and it was confirmed that the microorganism was a novel microorganism. The mycological properties obtained are shown in the table below.
【０００８】[0008]
【表１】 [Table 1]
【０００９】[0009]
【表２】 [Table 2]
【００１０】[0010]
【表３】 [Table 3]
【００１１】[0011]
【表４】 [Table 4]
【００１２】[0012]
【表５】 （＋）：ｗｅａｋ ｐｏｓｉｔｉｖｅ[Table 5] (+): Weak positive
【００１３】以上の結果よりＫＦ−１株は、Bergey's Ma
nual of Systematic Bacteriology,Vol.1,p214(1984)
に基づいてPseudomonas paucimobilisと同定された。Ｋ
Ｆ−２株については細胞壁の分析を行った。その結果、
ジアミノ酸はメソ−ジアミノピメリン酸であり、ミコー
ル酸が存在することがわかった。脂肪酸の構造は直鎖状
飽和脂肪酸とＣ18の分岐型不飽和脂肪酸であり、分岐型
の不飽和脂肪酸のＣ10の位置にメチル基のついたTuberc
ulostearic acidであった。このこと及び以上の結果か
らＫＦ−２株は、Bergey's Manual of Systematic Bac
teriology ,Vol.2,p1479〜1480(1989)に基づいてRhodoc
occus ruberと同定された。尚、ＫＦ−１株及びＫＦ−
２株について、キノン組成及びＧ＋Ｃ含有量を測定した
結果、ＫＦ−１株のキノン組成はユビキノン−８（組成
比９９％）であり、Ｇ＋Ｃ含有量は６２．０モル％であ
り、ＫＦ−２株のキノン組成はメナキノン−８（Ｈ２）
（組成比９８％）であり、Ｇ＋Ｃ含有量は６７．０モル
％であった。以上のＫＦ−１株はＦＥＲＭ ＢＰ−５１
２２として、ＫＦ−２株はＦＥＲＭＢＰ−５１２１とし
てそれぞれ寄託されている。From the above results, the KF-1 strain was isolated from Bergey's Ma
nual of Systematic Bacteriology, Vol.1, p214 (1984)
On the basis of Pseudomonas paucimobilis. K
For the F-2 strain, cell wall analysis was performed. as a result,
The diamino acid was meso-diaminopimelic acid, and it was found that mycolic acid was present. Structure of the fatty acid is a straight chain saturated fatty acids and branched unsaturated fatty acids of C 18, Tuberc equipped with a methyl group at position to C 10 of the branched unsaturated fatty acids
ulostearic acid. From this and the above results, the KF-2 strain was obtained from Bergey's Manual of Systematic Bac.
Rhodoc based on teriology, Vol. 2, pp. 1479-1480 (1989)
occus ruber. In addition, KF-1 strain and KF-
For 2 strains, the results of measurement of quinone composition and G + C content, quinone composition of KF-1 strain is a ubiquinone-8 (composition ratio 99%), G + C content Oh <br/> at 62.0 mol% The quinone composition of the KF-2 strain was menaquinone-8 (H2)
(Composition ratio 98%), and the G + C content is 67.0 mol.
% . The above KF-1 strain is FERM BP-51
No. 22 and the KF-2 strain has been deposited as FERMBP-5121, respectively.
【００１４】本発明の微生物は、多環芳香族化合物を分
解する新規な微生物である。本発明でいう多環芳香族化
合物は、２個の芳香族環を有する芳香族類であり、芳香
族環が酸素原子、硫黄原子、或いは窒素原子と複素環を
形成する化合物も含まれる。本発明の微生物によって効
率良く分解される多環芳香族化合物としては、ビフェニ
ル、ジベンゾフラン、ジベンゾチオフェン、フルオレ
ン、カルバゾール、フェナントレン、アントラセン、ピ
レン等及びこれらの置換体が挙げられる。本発明の排水
処理方法は、上記の新規な微生物（菌株）を単独で又は
併用して活性汚泥方式で行う点が特徴であり、適用され
る排水は多環芳香族化合物を含有する排水である。従っ
て、上記の多環芳香族化合物を含む排水の処理に本発明
は特に有効である。The microorganism of the present invention is a novel microorganism that degrades polycyclic aromatic compounds. The polycyclic aromatic compound referred to in the present invention is an aromatic having two aromatic rings, and includes a compound in which an aromatic ring forms a heterocyclic ring with an oxygen atom, a sulfur atom, or a nitrogen atom. Examples of the polycyclic aromatic compound that is efficiently decomposed by the microorganism of the present invention include biphenyl, dibenzofuran, dibenzothiophene, fluorene, carbazole, phenanthrene, anthracene, pyrene and the like, and substituted products thereof. The wastewater treatment method of the present invention is characterized in that the novel microorganism (strain) is used alone or in combination with an activated sludge method, and the wastewater to be applied is wastewater containing a polycyclic aromatic compound. . Therefore, the present invention is particularly effective for treating wastewater containing the above polycyclic aromatic compound.
【００１５】ところで、石油精製工場やコークス工場排
水には、多環芳香族化合物と共に高濃度のフェノールが
含まれている。従って、本発明の多環芳香族化合物分解
菌ＫＦ−１株及びＫＦ−２株は、生物毒性の高いフェノ
ールに対し耐性能力を有することも必要である。本発明
者等は、ＫＦ−１株及びＫＦ−２株に耐フェノール性を
付与すべく検討を重ね、これらの対数増殖期の培養菌体
に紫外線を照射して、或いは該培養菌体をエチルメタン
スルフォネート等の物質で処理する等によって突然変異
を起こさせ、得られた変異株をフェノール添加培地で馴
養することにより、ＫＦ−１株及びＫＦ−２株に耐フェ
ノール性が付与されることを見出した。このような耐フ
ェノール性が付与された改良ＫＦ−１株及びＫＦ−２株
を用いることにより高濃度フェノール中での多環芳香族
化合物の処理が可能となる。尚、これらの耐フェノール
性微生物の菌学的性質は、元のＫＦ−１株及びＫＦ−２
株と同一である。By the way, the effluent of a petroleum refinery or coke plant contains a high concentration of phenol together with a polycyclic aromatic compound. Therefore, the polycyclic aromatic compound-degrading bacteria KF-1 and KF-2 of the present invention also need to have the ability to withstand highly biotoxic phenol. The present inventors have repeatedly studied to impart phenol resistance to the KF-1 strain and the KF-2 strain, and irradiating the cultured cells in the logarithmic growth phase with ultraviolet light or subjecting the cultured cells to ethyl acetate. Mutation is caused by, for example, treatment with a substance such as methanesulfonate, and the resulting mutant strain is acclimated in a phenol-added medium, whereby phenol resistance is imparted to the KF-1 strain and the KF-2 strain. I found that. By using the improved KF-1 and KF-2 strains to which such phenol resistance has been imparted, it becomes possible to treat polycyclic aromatic compounds in high-concentration phenol. In addition, the mycological properties of these phenol-resistant microorganisms are based on the original KF-1 strain and KF-2.
Same as the strain.
【００１６】上記本発明の微生物はいずれも活性汚泥に
良好に定着し、馴養することにより活性汚泥の優先種と
なる性質を有しており、従って、本発明の微生物は活性
汚泥を利用する各種排水処理方法において特に有用であ
る。活性汚泥に対して本発明の微生物を定着させるに際
しては、それらの馴養にＳＲＴ（汚泥滞留時間／汚泥
齢）を、５日以上、好ましくは１０日以上取ることによ
り、本発明の微生物を活性汚泥に良好且つ安定に定着さ
せることができる。微生物を使用する活性汚泥方式によ
る排水処理方法自体は周知であり、上記の本発明の微生
物は周知のいずれの活性汚泥を用いる処理方法にも応用
することができるものであり、方法自体は特に限定され
るものではない。尚、上記の微生物の使用とは、その培
養物の使用、或はその培養処理物の使用も含むことはい
うまでもない。Each of the microorganisms of the present invention has the property of being well established in activated sludge and becoming a preferential species of activated sludge by acclimation. Therefore, the microorganism of the present invention can be used for various types of activated sludge. It is particularly useful in wastewater treatment methods. When the microorganisms of the present invention are established on activated sludge, the microorganisms of the present invention are activated sludge by taking SRT (sludge residence time / sludge age) for 5 days or more, preferably 10 days or more for their acclimation. Fixation can be performed satisfactorily and stably. The wastewater treatment method using the activated sludge method using microorganisms is well known, and the microorganism of the present invention can be applied to any known treatment method using activated sludge, and the method itself is particularly limited. It is not something to be done. Needless to say, the use of the above-mentioned microorganism includes the use of the culture or the use of the cultured product.
【００１７】[0017]
【実施例】次に実施例を挙げて本発明を更に具体的に説
明する。以下の実施例ではＫＦ−１株又はＫＦ−２株を
添加した表１に示す組成の培地に、フェナントレン、フ
ルオレン及びジベンゾチオフェンのいずれかを添加し
て、各試験時間（経過時間）による上記多環芳香族化合
物の濃度及び菌体量を測定した。更に具体的には、培地
１００ｍｌを５００ｍｌ容エーレンマイヤーフラスコに
入れ、多環芳香族化合物１００ｍｇを添加し、前培養し
た菌体を１ｍｌ添加し、３０℃、１８０ｒｐｍで所定時
間培養した。培養液を塩酸酸性にし、酢酸エチル１００
ｍｌ添加して抽出を行い、有機溶媒層を分取してガスク
ロマトグラフィーにより多環芳香族化合物を定量した。
又、菌体の量は吸光光度法により測定した。Next, the present invention will be described more specifically with reference to examples. In the following examples, any one of phenanthrene, fluorene and dibenzothiophene was added to a medium having the composition shown in Table 1 to which the KF-1 strain or the KF-2 strain had been added. The concentration of the aromatic ring compound and the amount of bacterial cells were measured. More specifically, 100 ml of the medium was placed in a 500 ml Erlenmeyer flask, 100 mg of the polycyclic aromatic compound was added, 1 ml of the pre-cultured cells were added, and the cells were cultured at 30 ° C. and 180 rpm for a predetermined time. The culture was acidified with hydrochloric acid, and ethyl acetate 100
Then, the organic solvent layer was separated and the amount of the polycyclic aromatic compound was determined by gas chromatography.
The amount of the cells was measured by an absorption spectrophotometry.
【００１８】[0018]
【表６】 [Table 6]
【００１９】実施例１ 本実施例ではＫＦ−１株によるフェナントレンの分解を
行った。得られた結果を表２に示す。Example 1 In this example, phenanthrene was decomposed by the KF-1 strain. Table 2 shows the obtained results.
【００２０】[0020]
【表７】 [Table 7]
【００２１】実施例２ ＫＦ−２株によりフルオレンを分解させた。結果を表３
に示す。Example 2 Fluorene was decomposed by the KF-2 strain. Table 3 shows the results
Shown in
【００２２】[0022]
【表８】 [Table 8]
【００２３】実施例３ ＫＦ−２株によりジベンゾチオフェンを分解させた。結
果を表４に示す。Example 3 Dibenzothiophene was decomposed by the KF-2 strain. Table 4 shows the results.
【００２４】[0024]
【表９】 [Table 9]
【００２５】実施例４ ＫＦ−１株によりフルオレンを分解させた。結果を表５
に示す。Example 4 Fluorene was decomposed by the KF-1 strain. Table 5 shows the results
Shown in
【００２６】[0026]
【表１０】 [Table 10]
【００２７】実施例５ ＫＦ−１株によりジベンゾチオフェンを分解させた。結
果を表６に示す。Example 5 Dibenzothiophene was decomposed by the KF-1 strain. Table 6 shows the results.
【００２８】[0028]
【表１１】 [Table 11]
【００２９】実施例６ 本実施例では、ＫＦ−１株を添加した活性汚泥によるフ
ェナントレンの分解除去例を示す。Example 6 In this example, an example of decomposing and removing phenanthrene by activated sludge to which the KF-1 strain was added will be described.
【００３０】（１）活性汚泥の調製 下水処理場の活性汚泥にＫＦ−１株を添加し、定着及び
馴養して使用した。活性汚泥の初期ＭＬＳＳを２０００
ｍｇ／ｌに調整し、ＫＦ−１株を初期菌体濃度１００ｍ
ｇ／ｌ（乾燥濃度）になるように添加した。 （２）合成排水及び負荷条件 下記表７に記載した組成のＳＧＰ合成排水を使用した。
ＳＧＰ合成排水を０．１ｋｇ−ＢＯＤ／ｋｇ−ＭＬＳＳ
・ｄａｙ、フェナントレンを０．０５ｋｇ／ｋｇ−ＭＬ
ＳＳ・ｄａｙの負荷条件となるように両者を同時に活性
汚泥に添加した。(1) Preparation of Activated Sludge KF-1 strain was added to activated sludge in a sewage treatment plant, and used after fixing and acclimatization. 2,000 initial MLSS of activated sludge
mg / l, and the KF-1 strain was adjusted to an initial cell concentration of 100 m.
g / l (dry concentration). (2) Synthetic wastewater and loading conditions SGP synthetic wastewater having the composition shown in Table 7 below was used.
0.1kg-BOD / kg-MLSS of SGP synthetic wastewater
・ Day, phenanthrene 0.05 kg / kg-ML
Both were added to the activated sludge at the same time so as to satisfy the loading condition of SS day.
【００３１】（３）ＳＲＴ（汚泥滞留時間／汚泥齢） ＳＲＴは、５、１０、２０、３０及び５０日で行った。 （４）処理方法 １リットル三角フラスコに（２）で調製した排水を５０
０ｍｌ入れ、室温で振とう培養を所定時間（１０、１
５、２０及び２５時間）行い、その後４時間静置して汚
泥を沈降分離させた。処理水を適当量引き抜き、処理水
中のフェナントレンの分析を行った。又、ＭＬＳＳ中の
フェナントレンの分析を行った。結果を表８及び図１
（処理時間とＭＬＳＳ中のフェナントレン濃度の関係）
に示す。ＳＲＴが１０日間以上では、いずれの処理時間
においても、処理水中のフェナントレン濃度は１ｍｇ／
ｌ以下（処理前５０ｍｇ／ｌ）であった。(3) SRT (sludge residence time / sludge age) SRT was performed for 5, 10, 20, 30, and 50 days. (4) Treatment method The wastewater prepared in (2) was placed in a 1-liter Erlenmeyer flask with 50
0 ml and shake culture at room temperature for a predetermined time (10, 1
(5, 20 and 25 hours), and then allowed to stand for 4 hours to settle and separate sludge. An appropriate amount of the treated water was withdrawn, and phenanthrene in the treated water was analyzed. In addition, phenanthrene in MLSS was analyzed. The results are shown in Table 8 and FIG.
(Relation between processing time and phenanthrene concentration in MLSS)
Shown in When the SRT is 10 days or more, the phenanthrene concentration in the treated water is 1 mg /
1 (50 mg / l before treatment).
【００３２】[0032]
【表１２】 [Table 12]
【００３３】[0033]
【表１３】 [Table 13]
【００３４】表８及び図１より、ＳＲＴを５日以上、好
ましくは１０日以上取れば、フェナントレンはＭＬＳＳ
中にも蓄積されず、安定した処理が行われていることが
わかる。従って、ＫＦ−１株はＳＲＴを５日以上、好ま
しくは１０日以上取ればＭＬＳＳ中に安定して定着する
と思われる。According to Table 8 and FIG. 1, if the SRT is taken for 5 days or more, preferably for 10 days or more, phenanthrene will be MLSS.
It is understood that stable processing is being performed without being accumulated in the inside. Therefore, it seems that the KF-1 strain stably colonizes the MLSS when the SRT is taken for 5 days or more, preferably 10 days or more.
【００３５】実施例７ 本実施例ではＫＦ−１株及びＫＦ−２株に耐フェノール
性を付与する方法及びその結果を示す。 （１）対数増殖期の決定 Nutrient Broth（ＮＢ）培地で前培養を行った上記株の
前培養液１ｍｌをＮＢ培地１００ｍｌに殖菌し、３０℃
で振とう培養を行った。この培養液の濁度、生菌数及び
ＡＴＰ量の経時変化を測定し、対数増殖期を決定した。
ＫＦ−１株及びＫＦ−２株の対数増殖期は、それぞれ４
〜１０時間及び６〜１４時間であった。 （２）紫外線照射条件（照射距離及び照射時間）の決定 紫外線照射による突然変異株の取得は、通常、対数増殖
期の菌体を用い、高い死滅率（９９．９９％程度）とな
る条件で行われる。対数増殖期の菌体を適当濃度（１０
-2〜１０-3）に希釈し、紫外線を所定距離（１０〜５０
ｃｍ）、所定時間（１０〜１８０秒）照射し、生菌数を
測定して最適条件を決定した。紫外線照射は東芝ランプ
ＧＬ１５０（東芝社製）で行った。Example 7 In this example, a method for imparting phenol resistance to the KF-1 strain and the KF-2 strain and the results thereof will be described. (1) Determination of logarithmic growth phase 1 ml of the pre-culture solution of the above strain, which had been pre-cultured in Nutrient Broth (NB) medium, was inoculated into 100 ml of NB medium and incubated at 30 ° C.
Shaking culture was performed. Changes in the turbidity, viable cell count, and ATP amount of this culture over time were measured, and the logarithmic growth phase was determined.
The logarithmic growth phase of the KF-1 and KF-2 strains was 4
-10 hours and 6-14 hours. (2) Determination of UV Irradiation Conditions (Irradiation Distance and Irradiation Time) Mutants by ultraviolet irradiation are usually obtained using a logarithmically growing bacterial cell under conditions that result in a high mortality (about 99.99%). Done. The cells in the logarithmic growth phase are grown at an appropriate concentration (10
-2 to 10 -3 ) and dilute the ultraviolet light at a predetermined distance (10 to 50
cm) for a predetermined time (10 to 180 seconds), and the viable cell count was measured to determine the optimal conditions. The ultraviolet irradiation was performed with a Toshiba lamp GL150 (manufactured by Toshiba Corporation).
【００３６】以上の結果を表９及び表１０に示す。ＫＦ
−１株の最適照射条件（菌体の死滅率が９９．９９％と
なる）は、照射距離１５〜３０ｃｍでは１０秒照射、５
０ｃｍでは３０秒照射である。又、ＫＦ−２株の最適照
射条件は、照射距離１０〜３０ｃｍでは１０秒照射、５
０ｃｍでは６０秒照射である。The above results are shown in Tables 9 and 10. KF
The optimal irradiation conditions for the -1 strain (the killing rate of the bacterial cells is 99.99%) are as follows.
At 0 cm, irradiation is for 30 seconds. The optimum irradiation conditions for the KF-2 strain are 10 seconds irradiation at an irradiation distance of 10 to 30 cm, 5 seconds irradiation,
At 0 cm, irradiation is for 60 seconds.
【００３７】[0037]
【表１４】 [Table 14]
【００３８】[0038]
【表１５】 [Table 15]
【００３９】（３）紫外線照射変異菌の取得 対数増殖期のＫＦ−１株及びＫＦ−２株を０．１ｍｌ、
平板培地に塗抹し、（２）で決定したそれぞれの最適条
件で紫外線を照射した。 （４）フェノール耐性の改良 紫外線照射したコロニーをフェノール濃度５００ｐｐｍ
のフェナントレン培地（試験管）に添加し菌体の増殖を
見た。増殖の見られた試験管から希釈平板を行い、これ
に上記の最適条件で紫外線を照射した。出現したコロニ
ーをフェノール濃度６００ｐｐｍのフェナントレン培地
で培養した。この操作を繰り返し、フェノール濃度を１
００ｐｐｍずつ上げていった。(3) Obtaining Ultraviolet Irradiated Mutants 0.1 ml of KF-1 strain and KF-2 strain in logarithmic growth phase
The plate medium was smeared and irradiated with ultraviolet light under the respective optimum conditions determined in (2). (4) Improvement of phenol resistance The colonies irradiated with ultraviolet light were subjected to phenol concentration of 500 ppm
Was added to a phenanthrene medium (test tube). A dilution plate was prepared from the test tube in which growth was observed, and irradiated with ultraviolet light under the above-mentioned optimum conditions. The appeared colonies were cultured in a phenanthrene medium having a phenol concentration of 600 ppm. This operation was repeated until the phenol concentration reached 1
It was increased by 00 ppm.
【００４０】ＫＦ−１株は、フェナントレン培地でのフ
ェノール耐性は７００ｐｐｍであったが、紫外線照射及
び馴養操作を繰り返す改良によりフェノール耐性は１４
００ｐｐｍまで向上した。ＫＦ−２株でも同様に、フェ
ノール耐性が著しく向上した。The KF-1 strain had a phenol resistance of 700 ppm in the phenanthrene medium, but had a phenol resistance of 14 ppm due to the improvement of repeated irradiation and adaptation.
Improved to 00 ppm. Similarly, the phenol resistance was remarkably improved in the KF-2 strain.
【００４１】[0041]
【発明の効果】以上の本発明によれば、新規な微生物を
用いることによって難分解性の多環芳香族化合物を分解
除去することが可能である。According to the present invention described above, it is possible to decompose and remove hardly decomposable polycyclic aromatic compounds by using a novel microorganism.
【図１】 実施例６の試験結果を示す図である。FIG. 1 is a diagram showing test results of Example 6.
───────────────────────────────────────────────────── フロントページの続き (51)Int.Cl.7 識別記号 ＦＩ Ｃ１２Ｒ 1:38） (Ｃ１２Ｎ 1/20 Ｃ１２Ｒ 1:01） 審査官 内田 俊生 (58)調査した分野(Int.Cl.7，ＤＢ名) C12N 1/20 C12N 1/26 C02F 3/34 ＢＩＯＳＩＳ（ＤＩＡＬＯＧ) ＷＰＩ（ＤＩＡＬＯＧ)──────────────────────────────────────────────────の Continuing on the front page (51) Int.Cl. 7 identification symbol FI C12R 1:38) (C12N 1/20 C12R 1:01) Examiner Toshio Uchida (58) Investigated field (Int.Cl. 7 , DB name) C12N 1/20 C12N 1/26 C02F 3/34 BIOSIS (DIALOG) WPI (DIALOG)
Claims (7)
するPseudomonas paucimobilis菌及びRhodococcus rube
r菌。1. Pseudomonas paucimobilis and Rhodococcus rube which degrade a polycyclic aromatic compound having a heterocyclic ring
r fungus.
nas paucimobilis４２１Ｙ株（ＦＥＲＭ ＢＰ−５１２
２）であり、Rhodococcus ruber菌がRhodococcus ruber
ＴＡ ０９０２株（ＦＥＲＭ ＢＰ−５１２１）であ
る請求項１に記載の複素環を有する多環芳香族化合物を
分解する菌。2. The method of claim 1] Pseudomonas paucimobilis bacteria Pseudomo
nas paucimobilis 421Y strain (FERM BP-512)
2) and the Rhodococcus ruber bacterium is
The bacterium that degrades a polycyclic aromatic compound having a heterocycle according to claim 1 , which is a TA0902 strain (FERM BP-5121).
の複素環を有する多環芳香族化合物を分解する菌。3. The bacterium that degrades the polycyclic aromatic compound having a heterocycle according to claim 1, which is resistant to phenol.
香族化合物分解菌に突然変異を起こさせ、生成変異株を
フェノール添加培地で馴養することを特徴とする複素環
を有する多環芳香族化合物分解菌に耐フェノール性を付
与する方法。4. A polycyclic heterocyclic ring having a heterocyclic ring, wherein the heterocyclic-degrading polycyclic aromatic compound-degrading bacterium according to claim 1 is mutated, and the resulting mutant strain is adapted to a phenol-containing medium. A method of imparting phenol resistance to aromatic compound degrading bacteria.
するPseudomonas paucimobilis菌及び／又はRhodococcu
s ruber菌を使用することを特徴とする多環芳香族化合
物含有排水の処理方法。5. A Pseudomonas paucimobilis bacterium and / or Rhodococcu which degrade a polycyclic aromatic compound having a heterocyclic ring.
A method for treating wastewater containing polycyclic aromatic compounds, characterized by using s ruber bacteria.
し、ＳＲＴ１０日以上で馴養することを特徴とする菌株
の馴養方法。6. A method of acclimating a strain, comprising adding the fungus according to claim 1 to activated sludge and acclimating for 10 days or more by SRT.
養微生物を使用することを特徴とする請求項５に記載の
排水の処理方法。7. The method for treating wastewater according to claim 5 , wherein the microorganism acclimated by the method according to claim 6 is used.
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| JP8711596A Division JP2922842B2 (en) | 1995-06-16 | 1996-03-18 | Method for imparting phenol resistance to microorganisms that degrade polycyclic aromatic compounds and method for treating polycyclic aromatic compound-containing wastewater using the phenol resistant microorganisms |
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| Publication Number | Publication Date |
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