JP2008061624A - New microorganism and liquid-waste disposal method using the same - Google Patents
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本発明は、高塩濃度下で有機化合物を分解可能な新規微生物、およびこれを用いた廃液処理方法に関するものである。 The present invention relates to a novel microorganism capable of decomposing an organic compound under a high salt concentration, and a waste liquid treatment method using the same.
種々の工業原料を効率的に製造する場合、酸塩化物とアルカリ金属化合物との反応や、有機塩素化合物とＮａ化合物との反応など、脱ＮａＣｌによる反応が有効な場合が多い。しかし、上記の製法では大量のＮａＣｌが副生するため、それを含む廃液処理を低コストで効率的に行なう必要がある。また、前記反応工程で生成した合成物質が有害物である場合、前記廃液中に合成物質が微量残留すると、高濃度食塩を含む廃液中で前記合成物質を分解処理するプロセス（無害化プロセス）も構築しなければならない。 When various industrial raw materials are efficiently produced, a reaction using deNaCl is often effective, such as a reaction between an acid chloride and an alkali metal compound or a reaction between an organic chlorine compound and a Na compound. However, since a large amount of NaCl is produced as a by-product in the above-described production method, it is necessary to efficiently perform waste liquid treatment including it at a low cost. In addition, when the synthetic substance generated in the reaction step is a harmful substance, a process (detoxification process) of decomposing the synthetic substance in the waste liquid containing high-concentration sodium chloride if a small amount of the synthetic substance remains in the waste liquid. Must be built.
上記の無害化プロセスについて、メタリルアルコールの製造法を例にして具体的に説明する。メタリルアルコールは、周知のとおり、医薬、農薬または各種化学品などの原料として有用である。従来から、メタリルアルコールの製造法としては、メタリルクロライドをＮａＯＨと反応させ、加水分解反応によりメタリルクロライド中のクロル基を水酸基で置換する方法が知られている（例えば、特許文献１〜４参照）。
The above detoxification process will be specifically described with reference to a method for producing methallyl alcohol. As is well known, methallyl alcohol is useful as a raw material for pharmaceuticals, agricultural chemicals or various chemicals. Conventionally, as a method for producing methallyl alcohol, there is known a method in which methallyl chloride is reacted with NaOH, and a chloro group in methallyl chloride is substituted with a hydroxyl group by a hydrolysis reaction (for example,
しかし、上記従来の方法では、大量のＮａＣｌが副生するため、廃液中には高濃度食塩が含まれる。また、本発明者の検討によれば、反応生成物であるメタリルアルコールと前記廃液とを分離した場合、該廃液中にメタリルアルコールが微量（数％程度）含まれてしまう。したがって、上記従来の方法でメタリルアルコールを製造する場合、廃液中に微量含まれるメタリルアルコールを高塩濃度下で分解処理する無害化プロセスを確立する必要がある。 However, in the conventional method described above, a large amount of NaCl is by-produced, and thus the waste liquid contains high-concentration sodium chloride. Further, according to the study of the present inventor, when methallyl alcohol, which is a reaction product, and the waste liquid are separated, a trace amount (about several percent) of methallyl alcohol is contained in the waste liquid. Therefore, when producing methallyl alcohol by the above conventional method, it is necessary to establish a detoxification process for decomposing methallyl alcohol contained in a small amount in the waste liquid under a high salt concentration.
上記した組成からなる廃液を処理するにあたり、高塩濃度下でメタリルアルコールやその他の有機化合物を分解可能な微生物の出現が望まれるが、このような性質を十分に備えた微生物は未だ発見されていない。 In the treatment of waste liquids having the above-mentioned composition, it is desired that microorganisms capable of decomposing methallyl alcohol and other organic compounds under high salt concentration are desired. However, microorganisms having such properties have not yet been discovered. Not.
本発明は、上記事情に鑑みてなされたものであり、その目的は、高塩濃度下で有機化合物を分解可能な新規微生物、および該微生物を用いて有機化合物の製造工程で副生する廃液中に含まれる有機化合物を分解処理する方法を提供することにある。 The present invention has been made in view of the above circumstances, and its purpose is in a novel microorganism capable of decomposing an organic compound under a high salt concentration, and in a waste liquid by-produced in the production process of the organic compound using the microorganism. Is to provide a method for decomposing an organic compound contained in the above.
本発明者は、上記課題を解決するため鋭意検討した結果、ハロモナス（Halomonas）属に属する菌の中に、高塩濃度下で有機化合物を分解可能な菌株を見出し、本発明を完成した。 As a result of intensive studies to solve the above problems, the present inventor found a strain capable of degrading an organic compound under a high salt concentration among bacteria belonging to the genus Halomonas, and completed the present invention.
すなわち、本発明の要旨は以下のとおりである。
〔１〕 高塩濃度下で有機化合物を分解可能なハロモナス（Halomonas）sp. 2Ｍ 株（FERM P-21006）、
〔２〕 高塩濃度下で有機化合物を分解可能なハロモナス（Halomonas）sp. 2Ｍ 株（FERM P-21006）の菌体または菌体培養液を、高濃度食塩を含む廃液と接触させ、該廃液中に含まれる有機化合物を分解処理することを特徴とする、高濃度食塩を含む廃液の処理方法、
〔３〕 廃液中に含まれる有機化合物が、有機塩素化合物中のクロル基を水酸基で置換してアルコールを得る過程で副生した高濃度食塩中に含まれる微量の前記アルコールである、前記〔２〕記載の方法、
〔４〕 アルコールがメタリルアルコールである、前記〔３〕記載の方法。
That is, the gist of the present invention is as follows.
[1] Halomonas sp. 2M strain (FERM P-21006) capable of degrading organic compounds under high salt concentration
[2] A cell or cell culture solution of Halomonas sp. 2M strain (FERM P-21006) capable of degrading organic compounds under high salt concentration is brought into contact with a waste solution containing high-concentration sodium chloride. A method for treating a waste liquid containing high-concentration sodium chloride, characterized by decomposing an organic compound contained therein,
[3] The organic compound contained in the waste liquid is a small amount of the alcohol contained in the high-concentration sodium chloride produced as a by-product in the process of obtaining an alcohol by substituting the chloro group in the organic chlorine compound with a hydroxyl group. The method described above,
[4] The method according to [3] above, wherein the alcohol is methallyl alcohol.
本発明のハロモナス（Halomonas）sp. 2Ｍ 株（FERM P-21006）によれば、高塩濃度下で有機化合物を分解することができる。また、該微生物を用いて有機化合物の製造工程で副生する廃液中に含まれる有機化合物を効率よく分解処理することができる。 According to the Halomonas sp. 2M strain (FERM P-21006) of the present invention, an organic compound can be decomposed under a high salt concentration. Moreover, the organic compound contained in the waste liquid byproduced in the manufacturing process of an organic compound can be efficiently decomposed using the microorganism.
１．目的微生物のスクリーニング
本発明者は、南極で採取した好気・好塩サンプル約１００個を被検サンプルとし、このうち、高塩濃度下（２Ｍ）で有機化合物（メタリルアルコール）を分解する性質を示す１株（2Ｍ 株）を単離した。
1. Screening of target microorganisms The present inventor used about 100 aerobic and halophilic samples collected in Antarctica as test samples, and among them, the property of decomposing organic compounds (methallyl alcohol) under high salt concentration (2M). 1 strain (2M strain) was isolated.
２．分類上の位置
2Ｍ 株の１６Ｓ ｒＲＮＡ配列相同性から判断した結果、2Ｍ 株はハロモナス（Halomonas）属に属する菌であると同定した。一方、種については、１６Ｓ ｒＲＮＡ配列相同性より既知の種とは一致しなかった（表１参照）。具体的には、Halomonas sp.M29 、Halomonas sp.M70 、Halomonas sp.M69 、Halomonas sp.HB-N 及びHalomonas sp.SB J85 に対してそれぞれ９７％の相同性を示した。そこで、2Ｍ 株はハロモナス（Halomonas）属に属する新菌種と同定し、ハロモナス（Halomonas）sp. 2Ｍ 株とした。ハロモナス（Halomonas）sp. 2Ｍ 株は独立行政法人産業技術総合研究所特許生物寄託センターにFERM P-21006として寄託されている。
2. Classification position
Judging from the 16S rRNA sequence homology of the 2M strain, the 2M strain was identified as a bacterium belonging to the genus Halomonas. On the other hand, the species did not match the known species due to 16S rRNA sequence homology (see Table 1). Specifically, it showed 97% homology with Halomonas sp. M29, Halomonas sp. M70, Halomonas sp. M69, Halomonas sp. HB-N and Halomonas sp. SB J85, respectively. Therefore, the 2M strain was identified as a new strain belonging to the genus Halomonas, and was designated as Halomonas sp. 2M strain. Halomonas sp. 2M strain has been deposited as FERM P-21006 at the Patent Organism Depositary, National Institute of Advanced Industrial Science and Technology.
３．廃液中に含まれる有機化合物の分解処理方法
本発明では、2Ｍ 株の菌体または菌体培養液を、高濃度食塩を含む廃液と接触させ、該廃液中に含まれる有機化合物を分解処理することで、前記廃液が処理される。
3. Method for Decomposing Organic Compounds Contained in Waste Liquid In the present invention, 2M strain cells or cell culture solution is brought into contact with a waste liquid containing high-concentration salt to decompose the organic compounds contained in the waste liquid. Then, the waste liquid is treated.
本発明でいう有機化合物とは、高塩濃度下で2Ｍ 株で分解可能な化合物をいい、例えば、アリルアルコールや有機カルボン酸などが例示され、典型的にはメタリルアルコールが例示される。高塩濃度としては特に限定されないが、通常は１．５〜３．５Ｍ、好ましくは２〜３Ｍをいう。また、前記有機化合物が含まれる廃液とは、例えば、酸塩化物とアルカリ金属化合物との反応や、有機塩素化合物とＮａ化合物との反応など脱ＮａＣｌを伴なう反応において副生するＮａＣｌを含むものをいう。通常、この廃液中には目的製造物が微量（〜数％）含まれている。 The organic compound referred to in the present invention refers to a compound that can be decomposed by a 2M strain under a high salt concentration, and examples thereof include allyl alcohol and organic carboxylic acid, and typically methallyl alcohol. Although it does not specifically limit as high salt concentration, Usually, 1.5-3.5M, Preferably 2-3M is said. The waste liquid containing the organic compound includes, for example, NaCl produced as a by-product in a reaction involving deNaCl such as a reaction between an acid chloride and an alkali metal compound or a reaction between an organic chlorine compound and a Na compound. Say things. Usually, the waste product contains a trace amount (up to several percent) of the target product.
前記廃液中に含まれる有機化合物の分解処理には、2Ｍ 株の菌体培養液のほか、該培養液を遠心分離等の集菌操作によって得られる菌体を用いることができる。分解処理は、前記菌株を前記廃液中に添加して行なえばよく、分解条件は廃液の量や有機化合物の濃度などに応じて適宜設定すればよい。 In the decomposition treatment of the organic compound contained in the waste liquid, in addition to the 2M strain cell culture solution, cells obtained by collecting the culture solution by centrifugation or the like can be used. The decomposition treatment may be performed by adding the strain to the waste liquid, and the decomposition conditions may be appropriately set according to the amount of the waste liquid and the concentration of the organic compound.
以下、試験例などにより本発明をさらに詳しく説明するが、本発明はこれらに限定されるものではない。 Hereinafter, the present invention will be described in more detail with reference to test examples and the like, but the present invention is not limited thereto.
１． 目的微生物のスクリーニング
南極で採取した好気・好塩サンプル約１００個をそれぞれ表２に示すスクリーニング用培地５ｍｌに少量添加し、３０℃で振とう培養して菌が生育するかどうかを観察した。そして、菌の生育が見られたサンプルについては、表３に示す培地に植え継ぎをして菌が生育するかどうかを観察した。
1. Screening of target microorganisms About 100 aerobic and halophilic samples collected in Antarctica were added in small amounts to 5 ml of the screening medium shown in Table 2, respectively, and cultured at 30 ° C. with shaking to observe whether the bacteria grew. And about the sample in which the growth of the microbe was seen, it transplanted to the culture medium shown in Table 3, and observed whether the microbe grew.
表３から、メタリルアルコールを添加した培地Ａと添加しない培地Ｂとの生育を比べると、ほとんどのサンプルで培地Ｂの方が生育がよかった。また、酵母エキスを添加しない培地Ｃでは全てのサンプルで菌の生育が見られなかった。このことから、生えてきた菌は酵母エキスを炭素源とし、メタリルアルコールは資化していないと考えられる。しかし、表３のサンプル中、サンプルNo.198（以下、「2Ｍ 株」という）（船底池底の泥サンプル）は、培地Ｂよりも培地Ａの方が生育がよかった。このことから、2Ｍ 株は、酵母エキス中の成分を必須としながらも、メタリルアルコールを資化している可能性があると考えられる。 From Table 3, when the growth of medium A to which methallyl alcohol was added was compared to medium B to which methallyl alcohol was not added, medium B grew better for most samples. In addition, in the medium C to which no yeast extract was added, no bacterial growth was observed in all samples. From this, it is considered that the bacterium that has grown uses yeast extract as a carbon source and has not assimilated methallyl alcohol. However, among the samples in Table 3, Sample No. 198 (hereinafter referred to as “2M strain”) (mud sample at the bottom of the bottom pond) grew more in Medium A than in Medium B. From this, it is considered that the 2M strain may assimilate methallyl alcohol while requiring the components in the yeast extract.
２． 2Ｍ 株のメタリルアルコール資化性の検討
メタリルアルコール０〜０．５％、酵母エキス０〜０．０２％の範囲で７種類の培地を調製し、該倍地に2Ｍ 株を少量添加して２週間培養し、培養日数に対してＯＤ６６０の値をプロットした。得られた増殖曲線を図１に示す。図１から、（１）メタリルアルコールなしでは菌はわずかしか生えないが、メタリルアルコールを添加した場合、菌の生育が良好であること、（２）メタリルアルコール濃度が低いほど生育速度が速いこと、（３）酵母エキスなしでは菌が生えないこと、（４）酵母エキス濃度が高いほど生育速度が速いこと、が分かった。以上のことから、2Ｍ 株は酵母エキスを必須としながらも、メタリルアルコールを資化していることが確実と考えられる。
2. Examination of methallyl alcohol assimilation ability of 2M strains Prepare 7 kinds of mediums in the range of methallyl alcohol 0-0.5% and yeast extract 0-0.02%, and add a small amount of 2M strain to the medium. The OD 660 values were plotted against the number of culture days. The obtained growth curve is shown in FIG. From FIG. 1, (1) Bacteria grow only slightly without methallyl alcohol, but when methallyl alcohol is added, the growth of the bacterium is good, and (2) the lower the methallyl alcohol concentration, the higher the growth rate. It was found that (3) the yeast cannot grow without the yeast extract, and (4) the higher the yeast extract concentration, the faster the growth rate. From the above, it is considered that 2M strains assimilate methallyl alcohol while requiring yeast extract.
３． 2Ｍ 株の至適ＮａＣｌ濃度の検討
メタリルアルコール０．５％、酵母エキス０．０２％をそれぞれ含み、ＮａＣｌ濃度を１Ｍ，２Ｍ，３Ｍ，４Ｍ及び５Ｍ含むように調整した各培地に2Ｍ 株を添加して１週間培養し、培養日数に対してＯＤ６６０の値をプロットした。得られた増殖曲線を図２に示す。図２から、菌は、１ＭＮａＣｌでは少し生育したが、４Ｍ ＮａＣｌ以上では生育しなかった。一方、２Ｍおよび３Ｍ ＮａＣｌでは、菌の生育は良好だった。
3. Examination of optimal NaCl concentration of 2M strains 2M strains were added to each medium containing 0.5% methallyl alcohol and 0.02% yeast extract and adjusted to contain NaCl concentrations of 1M, 2M, 3M, 4M and 5M. After addition, the cells were cultured for 1 week, and the value of OD 660 was plotted against the number of culture days. The obtained growth curve is shown in FIG. From FIG. 2, the fungus grew slightly with 1M NaCl, but did not grow with 4M NaCl or more. On the other hand, with 2M and 3M NaCl, the growth of the fungi was good.
本発明に係る微生物（ハロモナス（Halomonas）sp. 2Ｍ 株（FERM P-21006））は、高塩濃度下で有機化合物を分解する廃液処理方法に広く利用することができる。 The microorganism (Halomonas sp. 2M strain (FERM P-21006)) according to the present invention can be widely used in a waste liquid treatment method for decomposing an organic compound under a high salt concentration.
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| JP2010536561A (en) * | 2007-08-23 | 2010-12-02 | ダウ グローバル テクノロジーズ インコーポレイティド | Methods, adapted microorganisms, compositions and equipment for the purification of industrial brine |
| CN104894016A (en) * | 2015-05-22 | 2015-09-09 | 国家海洋局第三海洋研究所 | Halomonas axialensis and application thereof in degrading of decabromodiphenyl ether |
| CN109929777A (en) * | 2019-02-26 | 2019-06-25 | 新疆农业科学院微生物应用研究所（中国新疆-亚美尼亚生物工程研究开发中心） | A kind of Halomonas strain H6, composition and its application in salt tolerant growth-promoting |
| JP2019162080A (en) * | 2018-03-20 | 2019-09-26 | 学校法人 東洋大学 | Method for decomposing lees of unrefined soy sauce and composition for decomposing lees of unrefined soy sauce |
| JP2019528169A (en) * | 2016-08-25 | 2019-10-10 | コベストロ、ドイチュラント、アクチエンゲゼルシャフトＣｏｖｅｓｔｒｏ Ｄｅｕｔｓｃｈｌａｎｄ Ａｇ | Formate-catalyzed reaction from high salinity environment by salt-tolerant halomonas species (Halomonas sp.) |
| JP2021506575A (en) * | 2017-12-19 | 2021-02-22 | コベストロ、ドイチュラント、アクチエンゲゼルシャフトＣｏｖｅｓｔｒｏ Ｄｅｕｔｓｃｈｌａｎｄ Ａｇ | Continuous methods for reducing the amount of organic compounds in wastewater |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2010536561A (en) * | 2007-08-23 | 2010-12-02 | ダウ グローバル テクノロジーズ インコーポレイティド | Methods, adapted microorganisms, compositions and equipment for the purification of industrial brine |
| CN104894016A (en) * | 2015-05-22 | 2015-09-09 | 国家海洋局第三海洋研究所 | Halomonas axialensis and application thereof in degrading of decabromodiphenyl ether |
| JP2019528169A (en) * | 2016-08-25 | 2019-10-10 | コベストロ、ドイチュラント、アクチエンゲゼルシャフトＣｏｖｅｓｔｒｏ Ｄｅｕｔｓｃｈｌａｎｄ Ａｇ | Formate-catalyzed reaction from high salinity environment by salt-tolerant halomonas species (Halomonas sp.) |
| JP2021506575A (en) * | 2017-12-19 | 2021-02-22 | コベストロ、ドイチュラント、アクチエンゲゼルシャフトＣｏｖｅｓｔｒｏ Ｄｅｕｔｓｃｈｌａｎｄ Ａｇ | Continuous methods for reducing the amount of organic compounds in wastewater |
| JP2019162080A (en) * | 2018-03-20 | 2019-09-26 | 学校法人 東洋大学 | Method for decomposing lees of unrefined soy sauce and composition for decomposing lees of unrefined soy sauce |
| JP7055356B2 (en) | 2018-03-20 | 2022-04-18 | 学校法人 東洋大学 | Method for Decomposing Soy Sauce Moromi Lees and Composition for Decomposing Soy Sauce Moromi Lees |
| CN109929777A (en) * | 2019-02-26 | 2019-06-25 | 新疆农业科学院微生物应用研究所（中国新疆-亚美尼亚生物工程研究开发中心） | A kind of Halomonas strain H6, composition and its application in salt tolerant growth-promoting |
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