JP5856749B2 - Biofilm remover - Google Patents
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- JP5856749B2 JP5856749B2 JP2011081601A JP2011081601A JP5856749B2 JP 5856749 B2 JP5856749 B2 JP 5856749B2 JP 2011081601 A JP2011081601 A JP 2011081601A JP 2011081601 A JP2011081601 A JP 2011081601A JP 5856749 B2 JP5856749 B2 JP 5856749B2
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- activated sludge
- separation
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- uronic acid
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/10—Biological treatment of water, waste water, or sewage
Landscapes
- Enzymes And Modification Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Separation Using Semi-Permeable Membranes (AREA)
- Activated Sludge Processes (AREA)
Description
本発明は、バイオフィルム除去剤に関する。本発明はまた、該バイオフィルム除去剤を用いたバイオフィルムの除去方法、膜分離方法、膜分離活性汚泥方法および有機性廃水の処理装置に関する。 The present invention relates to a biofilm remover. The present invention also relates to a biofilm removal method, a membrane separation method, a membrane separation activated sludge method, and an organic wastewater treatment apparatus using the biofilm remover.
人工、自然環境を問わず、固体表面と水とが接する場面では、必ずといっていいほどバイオフィルムが存在する。バイオフィルムとは、固体表面に吸着した微生物が、細胞外多糖などの有機高分子マトリックスを産出し、そのマトリックス内に入り込んだ微生物の群落である。バイオフィルムはあらゆる固体表面に発生し、種々の問題を引き起こす。例えば、水道管や船底での流体の摩擦抵抗の増加や、熱交換器の熱移動の減少、金属材料の腐食や、食品、医療機器の汚染などが挙げられる。そして河川水や海水、活性汚泥などの液体をろ過する膜分離においても、水相中に放出されたバイオフィルム中の有機高分子が、膜の目詰まりの原因となる。 Regardless of whether it is man-made or natural, there is always a biofilm where the surface of the solid comes into contact with water. A biofilm is a community of microorganisms in which microorganisms adsorbed on a solid surface produce an organic polymer matrix such as an extracellular polysaccharide and enter the matrix. Biofilms occur on any solid surface and cause various problems. For example, there is an increase in frictional resistance of fluids in water pipes and ship bottoms, a decrease in heat transfer of heat exchangers, corrosion of metal materials, and contamination of food and medical equipment. In membrane separation for filtering liquids such as river water, seawater, and activated sludge, the organic polymer in the biofilm released into the aqueous phase causes membrane clogging.
バイオフィルム内で増殖した微生物は、浮遊性の微生物に比べて、抗生物質や殺菌剤に対しての耐性がより強く、一度形成されると除去することは容易なことではない。バイオフィルムは次に述べる一連の基本的な過程を経て形成される：(i) 裸の固体表面へのイオン、有機物の吸着；(ii) (i)への微生物細胞の付着；(iii) 付着した細胞の増殖とそれにともなう細胞外多糖の生産；および(iv) 他の細菌、微生物も含めた共同体としてのバイオフィルムの成長。このようなバイオフィルムの形成過程に対し、常にそれを阻止する脱離作用が働いている。例えば、バイオフィルム近傍に水の流れがあれば、せん断力が働き、それによる脱離作用によって、水相中にバイオフィルムが放出される場合もある。このようにして固体表面へ付着したバイオフィルムや、水相中へ放出されたバイオフィルムは、いずれも前述のような種々の問題を引き起こす。 Microorganisms grown in biofilms are more resistant to antibiotics and fungicides than floating microorganisms and are not easy to remove once formed. Biofilms are formed through a series of basic processes as follows: (i) adsorption of ions and organic matter on bare solid surfaces; (ii) attachment of microbial cells to (i); (iii) attachment Cell proliferation and associated extracellular polysaccharide production; and (iv) growth of biofilms as a community including other bacteria and microorganisms. In the process of forming such a biofilm, a detaching action that always prevents it is working. For example, if there is a flow of water in the vicinity of the biofilm, a shearing force acts, and the biofilm may be released into the aqueous phase due to the detachment action. The biofilm adhered to the solid surface in this way and the biofilm released into the aqueous phase all cause various problems as described above.
このような問題に対して、例えば特許文献１には、プロテアーゼのグループから選ばれる少なくとも１種類の酵素と、エステラーゼのグループから選ばれる少なくとも１種類の酵素と、アミラーゼとを含む酵素混合物の作用と、アルカリ性の洗浄剤での洗浄を組み合わせた、バイオフィルムの除去方法が開示されている。 For example, Patent Document 1 discloses an action of an enzyme mixture containing at least one enzyme selected from the protease group, at least one enzyme selected from the esterase group, and amylase. A method for removing a biofilm is disclosed that combines washing with an alkaline detergent.
一方、特許文献２には、膜分離活性汚泥法において、生物処理槽（曝気槽）内に浸漬設置した第１分離手段をなす浸漬型膜分離装置で活性汚泥混合液を固液分離し、活性汚泥処理により生物処理槽内に蓄積される生物由来ポリマーを含むＣＯＤ（化学的酸素要求量）を、第２分離手段によって適時に活性汚泥混合液から固液分離して、生物処理槽内の活性汚泥量を高濃度に維持しつつ、活性汚泥混合液中の生物由来ポリマー量を低濃度に維持することを特徴とする、膜面に過剰なポリマーが付着することを防止する方法が開示されている。 On the other hand, in Patent Document 2, in the membrane separation activated sludge method, the activated sludge mixed solution is solid-liquid separated by an immersion type membrane separation apparatus that constitutes a first separation means immersed in a biological treatment tank (aeration tank). The COD (chemical oxygen demand) containing the bio-derived polymer accumulated in the biological treatment tank by the sludge treatment is solid-liquid separated from the activated sludge mixed solution in a timely manner by the second separation means, and the activity in the biological treatment tank is achieved. Disclosed is a method for preventing excessive polymer from adhering to the membrane surface, characterized in that the amount of biological polymer in the activated sludge mixed liquid is maintained at a low concentration while maintaining the amount of sludge at a high concentration. Yes.
また、特許文献３には、有機性廃水を膜分離活性汚泥法により処理する際に、膜の目詰まりの原因となる原核生物を捕食する微小動物を生物反応槽内に添加することにより、排水処理を安定して長時間継続実施する手段が開示されている。 Further, Patent Document 3 discloses that when organic wastewater is treated by a membrane separation activated sludge method, by adding a micro animal that prey on prokaryotes that cause clogging of the membrane to the biological reaction tank, Means for stably performing the processing for a long time is disclosed.
しかしながら、特許文献１に記載の方法では、酵素混合物の作用とアルカリ洗浄剤による洗浄の両方が必要であるなどの煩雑さがあった。また、特許文献２に記載の方法では、膜の目詰まりの原因となるポリマーの量を推測する手段としてＣＯＤ値を求め、代用しているが、ＣＯＤでは、膜の細孔を素通りできる有機物も検出してしまうという問題がある。さらに、特許文献３に記載の方法では、バイオフィルム等の生物由来ポリマーによる目詰まりには対処することができないという問題がある。 However, in the method described in Patent Document 1, there is a problem that both the action of the enzyme mixture and the washing with an alkaline detergent are necessary. Further, in the method described in Patent Document 2, the COD value is obtained and used as a means for estimating the amount of the polymer causing the clogging of the membrane. However, in the COD, an organic substance that can pass through the pores of the membrane is also used. There is a problem of detecting. Furthermore, the method described in Patent Document 3 has a problem that it cannot cope with clogging due to a bio-derived polymer such as a biofilm.
このような背景のもと、本発明は、バイオフィルム除去剤および除去方法を提供することを目的とする。 Under such background, an object of the present invention is to provide a biofilm remover and a removal method.
本発明者らは上記目的を達成するために鋭意検討した結果、バイオフィルムに対し、骨格に５〜１００％の割合でウロン酸ユニットを含む多糖類を分解することができる多糖類分解酵素を作用させると、バイオフィルムを除去できることを見出し、本発明を完成した。 As a result of intensive studies to achieve the above-mentioned object, the present inventors acted on a biofilm with a polysaccharide-degrading enzyme capable of degrading a polysaccharide containing a uronic acid unit at a ratio of 5 to 100% in the skeleton. As a result, it was found that the biofilm could be removed, and the present invention was completed.
すなわち、本発明は、多糖類分解酵素を含むバイオフィルム除去剤であって、前記多糖類分解酵素が、ウロン酸ユニットを全糖ユニットに対し５〜１００％含む多糖類を分解する酵素である、バイオフィルム除去剤に関する。
本発明は、また、前記多糖類分解酵素が、キサンタンガム分解酵素、ヒアルロン酸分解酵素、ジェランガム分解酵素およびポリガラクツロン酸分解酵素からなる群から選択される１種または複数種の酵素である、前記バイオフィルム除去剤に関する。
さらに、本発明は、前記バイオフィルム除去剤でバイオフィルムを処理する工程を含む、バイオフィルムの除去方法に関する。
さらに、本発明は、分離膜が設置された分離膜装置を用いて被処理水をろ過する工程を含む膜分離方法であって、前記分離膜を、前記バイオフィルム除去剤で処理して、膜差圧の上昇を抑制する工程を含む、膜分離方法に関する。
さらに、本発明は、活性汚泥槽の水相中のウロン酸ユニット濃度の経時的な変化を測定し、該ウロン酸ユニット濃度が３０ｍｇ／Ｌに達したときに、前記バイオフィルム除去剤を活性汚泥に添加する工程を含む、膜分離活性汚泥法に関する。
さらに、本発明は、微生物を含む活性汚泥を収容した、有機性廃水を生物処理する活性汚泥槽と、該活性汚泥槽中またはその後段に設置した、生物処理水を固液分離する分離膜装置と、該活性汚泥槽中の水相中のウロン酸ユニット濃度の経時的な変化を測定するウロン酸ユニット濃度測定手段と、該ウロン酸ユニット濃度が所定の濃度に達したときに、前記バイオフィルム除去剤で該分離膜装置を処理するために、バイオフィルム除去剤を添加するバイオフィルム除去剤添加手段と、を含む、膜分離活性汚泥法による有機性廃水の処理装置に関する。
That is, the present invention is a biofilm remover comprising a polysaccharide-degrading enzyme, wherein the polysaccharide-degrading enzyme is an enzyme that degrades a polysaccharide containing 5 to 100% of a uronic acid unit with respect to the total sugar units. The present invention relates to a biofilm remover.
In the present invention, the polysaccharide degrading enzyme is one or more kinds of enzymes selected from the group consisting of xanthan gum degrading enzyme, hyaluronic acid degrading enzyme, gellan gum degrading enzyme and polygalacturonic acid degrading enzyme. The present invention relates to a film remover.
Furthermore, this invention relates to the removal method of a biofilm including the process of processing a biofilm with the said biofilm removal agent.
Furthermore, the present invention is a membrane separation method comprising a step of filtering water to be treated using a separation membrane device in which a separation membrane is installed, wherein the separation membrane is treated with the biofilm remover, and the membrane The present invention relates to a membrane separation method including a step of suppressing an increase in differential pressure.
Furthermore, the present invention measures the change over time of the uronic acid unit concentration in the aqueous phase of the activated sludge tank, and when the uronic acid unit concentration reaches 30 mg / L, the biofilm remover is added to the activated sludge. The present invention relates to a membrane separation activated sludge method including a step of adding to the membrane.
Furthermore, the present invention provides an activated sludge tank containing activated sludge containing microorganisms for biologically treating organic wastewater, and a separation membrane device installed in or at the subsequent stage of the activated sludge tank for solid-liquid separation of biologically treated water And uronic acid unit concentration measuring means for measuring a change with time of the uronic acid unit concentration in the aqueous phase in the activated sludge tank, and when the uronic acid unit concentration reaches a predetermined concentration, the biofilm The present invention relates to a device for treating organic wastewater by a membrane separation activated sludge method, comprising: a biofilm remover adding means for adding a biofilm remover to treat the separation membrane device with a remover.
本発明のバイオフィルム除去剤を使用することにより、固体表面に付着したバイオフィルムおよび水相中に放出されたバイオフィルムを除去することができる。特にこのバイオフィルム除去剤を膜分離活性汚泥法において用いることにより、分離膜の目詰まりを防ぎ、膜分離活性汚泥法による有機性廃水の処理を長期間安定的に行うことができる。 By using the biofilm remover of the present invention, the biofilm attached to the solid surface and the biofilm released into the aqueous phase can be removed. In particular, by using this biofilm removing agent in the membrane separation activated sludge method, clogging of the separation membrane can be prevented, and the treatment of organic waste water by the membrane separation activated sludge method can be performed stably for a long period of time.
以下、本発明を実施するための形態（以下、「本実施の形態」という。）について詳細に説明する。尚、本発明は、以下の本実施の形態に限定されるものではなく、その要旨の範囲内で種々変形して実施することができる。また、図面における上下左右等の位置関係は、特に断らない限り、図面に示す位置関係に基づくものとし、図面の寸法比率は、図示の比率に限られるものではない。 Hereinafter, a mode for carrying out the present invention (hereinafter referred to as “the present embodiment”) will be described in detail. The present invention is not limited to the following embodiment, and can be implemented with various modifications within the scope of the gist. In addition, the positional relationship such as up, down, left, and right in the drawing is based on the positional relationship shown in the drawing unless otherwise specified, and the dimensional ratio in the drawing is not limited to the illustrated ratio.
（バイオフィルム除去剤）
本実施の形態に係るバイオフィルム除去剤について説明する。人工、自然環境を問わず、固体表面と水とが接する場面では、必ずといっていいほどバイオフィルムが存在する。バイオフィルムとは、固体表面に吸着した微生物が、細胞外多糖などの有機高分子マトリックスを産出し、そのマトリックス内に入り込んだ微生物の群落であり、嫌気性微生物および好気性微生物のいずれも存在する。バイオフィルムはあらゆる固体表面に発生し、種々の問題を引き起こす。例えば、廃水処理における活性汚泥槽などにおいても、バイオフィルムは形成される。本発明者らは、このバイオフィルムが、多糖類分解酵素、特にウロン酸ユニットを含有する多糖類を分解することができる酵素により、分解除去可能であることを見出した。なお、本発明者らは、後述の実施例に示すとおり、特定のバイオフィルムについて構成多糖の分析を実施したところ、ヘキソース：５０％、デオキシ糖：２０％、アミノ糖：２０％およびウロン酸：１０％であったが、このような構成のバイオフィルムであっても、特にウロン酸ユニットを含有する多糖類を分解することができる酵素により、分解除去可能であることを見出した。
(Biofilm remover)
The biofilm remover according to the present embodiment will be described. Regardless of whether it is man-made or natural, there is always a biofilm where the surface of the solid comes into contact with water. A biofilm is a community of microorganisms in which microorganisms adsorbed on a solid surface produce an organic polymer matrix such as extracellular polysaccharides, and both anaerobic and aerobic microorganisms exist. . Biofilms occur on any solid surface and cause various problems. For example, biofilms are also formed in activated sludge tanks for wastewater treatment. The present inventors have found that this biofilm can be decomposed and removed by a polysaccharide-degrading enzyme, particularly an enzyme capable of degrading a polysaccharide containing a uronic acid unit. In addition, as shown in Examples described later, the present inventors conducted analysis of constituent polysaccharides on specific biofilms. As a result, hexose: 50%, deoxy sugar: 20%, amino sugar: 20%, and uronic acid: Although it was 10%, it was found that even a biofilm having such a structure can be decomposed and removed by an enzyme capable of decomposing a polysaccharide containing a uronic acid unit.
本実施の形態に係るバイオフィルム除去剤でバイオフィルムを処理することにより、主にバイオフィルムを形成する高分子マトリックスが分解され、バイオフィルムを分解および／または除去することができる。バイオフィルムの処理は、例えば、固体表面に発生したバイオフィルムと本実施の形態に係るバイオフィルム除去剤を接触させることによって行うことができる。また、活性汚泥槽などにおいて水相中に放出されたバイオフィルムの場合には、バイオフィルムを含む水（例えば、活性汚泥槽）に本実施の形態に係るバイオフィルム除去剤を添加することによって、バイオフィルムの処理を行うことができる。 By treating the biofilm with the biofilm removing agent according to the present embodiment, the polymer matrix that mainly forms the biofilm is decomposed, and the biofilm can be decomposed and / or removed. The treatment of the biofilm can be performed, for example, by bringing the biofilm generated on the solid surface into contact with the biofilm removing agent according to the present embodiment. Further, in the case of a biofilm released into the aqueous phase in an activated sludge tank or the like, by adding the biofilm remover according to the present embodiment to water containing the biofilm (for example, activated sludge tank), Biofilm processing can be performed.
本実施の形態に係るバイオフィルム除去剤は、多糖類分解酵素を含むことを特徴とする。該酵素が分解する多糖類としては、でんぷんなどの中性多糖、キチンなどのアミノ多糖、ポリウロン酸などの酸性多糖、キサンタンガムなどのウロン酸ユニット含有多糖類などが挙げられ、好ましくはウロン酸ユニット含有多糖類である。 The biofilm remover according to the present embodiment includes a polysaccharide degrading enzyme. Examples of the polysaccharide that the enzyme decomposes include neutral polysaccharides such as starch, amino polysaccharides such as chitin, acidic polysaccharides such as polyuronic acid, uronic acid unit-containing polysaccharides such as xanthan gum, and preferably uronic acid unit-containing polysaccharides It is a polysaccharide.
ウロン酸ユニット含有多糖類としては、単糖を酸化して得られる誘導体のうち、主鎖の末端のヒドロキシメチル基（−ＣＨ2ＯＨ）がカルボキシル基（−ＣＯ2Ｈ）に変換されたカルボン酸であるウロン酸をその骨格に含む多糖類であれば特に限定されず、例えば、グルクロン酸、ガラクツロン酸、マンヌロン酸、イズロン酸等のウロン酸ユニットを含む単糖類を構成単位とする多糖類であるキサンタンガム、ヒアルロン酸、アルギン酸、ヘパリン、コンドロイチン硫酸、ジェランガムや、単一のウロン酸ユニットのみからなる高分子であるポリウロン酸であるポリガラクツロン酸が挙げられるが、これらに限定されない。好ましくは、本実施の形態に係るバイオフィルム除去剤に含まれる多糖類分解酵素が分解するウロン酸ユニット含有多糖類は、キサンタンガム、ジェランガム、ヒアルロン酸またはポリガラクツロン酸であり、より好ましくは、キサンタンガム、ジェランガムまたはヒアルロン酸であり、さらに好ましくは、キサンタンガムまたはジェランガムであり、特に好ましくはキサンタンガムである。尚、本実施の形態に係るバイオフィルム除去剤に含まれる多糖類分解酵素は、これらの多糖類の１種または複数種を分解するものであってもよい。 The uronic acid unit-containing polysaccharide is a carboxylic acid in which a hydroxymethyl group (—CH 2 OH) at the end of the main chain is converted to a carboxyl group (—CO 2 H) among derivatives obtained by oxidizing a monosaccharide. It is not particularly limited as long as it is a polysaccharide containing uronic acid in its skeleton. For example, it is a polysaccharide comprising a monosaccharide containing a uronic acid unit such as glucuronic acid, galacturonic acid, mannuronic acid, iduronic acid as a structural unit. Examples include, but are not limited to, xanthan gum, hyaluronic acid, alginic acid, heparin, chondroitin sulfate, gellan gum, and polygalacturonic acid that is a polyuronic acid that is a polymer composed of only a single uronic acid unit. Preferably, the uronic acid unit-containing polysaccharide decomposed by the polysaccharide-degrading enzyme contained in the biofilm removing agent according to the present embodiment is xanthan gum, gellan gum, hyaluronic acid or polygalacturonic acid, more preferably xanthan gum, Gellan gum or hyaluronic acid, more preferably xanthan gum or gellan gum, and particularly preferably xanthan gum. In addition, the polysaccharide degrading enzyme contained in the biofilm removing agent according to the present embodiment may be one that degrades one or more of these polysaccharides.
前記多糖類を分解できる酵素としては、実際のバイオフィルムの分子構造と似ているという点から、キサンタンガム分解酵素、ヒアルロン酸分解酵素、ジェランガム分解酵素およびポリガラクツロン酸分解酵素からなる群から選択される１種または複数種の酵素が好ましく、キサンタンガム分解酵素、ヒアルロン酸分解酵素およびジェランガム分解酵素からなる群から選択される１種または複数種の酵素がより好ましく、キサンタンガム分解酵素およびジェランガム分解酵素からなる群から選択される１種または複数種の酵素がさらに好ましく、キサンタンガム分解酵素が含まれることが特に好ましい。 The enzyme capable of degrading the polysaccharide is selected from the group consisting of xanthan gum degrading enzyme, hyaluronic acid degrading enzyme, gellan gum degrading enzyme and polygalacturonic acid degrading enzyme because it is similar to the molecular structure of an actual biofilm. One or more enzymes are preferred, one or more enzymes selected from the group consisting of xanthan gum degrading enzymes, hyaluronic acid degrading enzymes and gellan gum degrading enzymes are more preferred, and groups consisting of xanthan gum degrading enzymes and gellan gum degrading enzymes One or more kinds of enzymes selected from are more preferred, and it is particularly preferred that xanthan gum-degrading enzymes are included.
本実施の形態に係るバイオフィルム除去剤に含まれる多糖類分解酵素が分解するウロン酸ユニット含有多糖類は、バイオフィルムの分解効率を高めるという観点から、好ましくはウロン酸ユニットと他の糖ユニットとを含有する多糖類である。例えば、好ましくは、ウロン酸ユニット濃度が全糖ユニット濃度の５〜１００％であり、より好ましくは７〜１００％であり、さらに好ましくは１０〜７０％であり、特に好ましくは２０〜５０％である。このような多糖類としては、例えば、キサンタンガム（ウロン酸ユニットを全糖ユニットに対し２０％含有）、ヒアルロン酸（ウロン酸ユニットを全糖ユニットに対し５０％含有）、ジェランガム（ウロン酸ユニットを全糖ユニットに対し２５％含有）、ポリガラクツロン酸（ウロン酸ユニットを全糖ユニットに対し１００％含有）、が挙げられる。 The uronic acid unit-containing polysaccharide that is decomposed by the polysaccharide-degrading enzyme contained in the biofilm remover according to the present embodiment is preferably a uronic acid unit and other saccharide units, from the viewpoint of increasing the degradation efficiency of the biofilm. Is a polysaccharide containing For example, preferably, the uronic acid unit concentration is 5 to 100% of the total sugar unit concentration, more preferably 7 to 100%, still more preferably 10 to 70%, and particularly preferably 20 to 50%. is there. Examples of such polysaccharides include xanthan gum (containing 20% uronic acid units relative to all sugar units), hyaluronic acid (containing 50% uronic acid units relative to all sugar units), and gellan gum (all uronic acid units included). 25% of sugar units) and polygalacturonic acid (containing 100% of uronic acid units with respect to all sugar units).
ここで全糖ユニット濃度は、以下に示すフェノール硫酸法により測定できる。
１）試験管に試料水溶液または標準単糖水溶液を５００μＬ加える
２）５ｗｔ％フェノール水溶液を５００μＬ加え、撹拌する
３）濃硫酸を２．５ｍＬ加え、すぐに１０秒間激しく撹拌する
４）３０℃水浴中に２０分以上放置する
５）分光光度計で４９０ｎｍの吸収を測定し、既知濃度の標準単糖で作製した検量線から濃度を求める
尚、ウロン酸ユニット濃度は、後述の手法を用いて測定することができる。
Here, the total sugar unit concentration can be measured by the phenol sulfuric acid method shown below.
1) Add 500 μL of sample aqueous solution or standard monosaccharide aqueous solution to the test tube 2) Add 500 μL of 5 wt% phenol aqueous solution and stir 3) Add 2.5 mL of concentrated sulfuric acid and immediately stir vigorously for 10 seconds 4) In a 30 ° C. water bath 5) Measure the absorption at 490 nm with a spectrophotometer and obtain the concentration from a calibration curve prepared with a standard monosaccharide of known concentration. The uronic acid unit concentration is measured using the method described below. be able to.
本実施の形態に係るバイオフィルム除去剤としては、例えば、ウロン酸ユニットを含む多糖類のみを炭素源として含む培地で培養可能な細菌コロニーを培養した培養液のろ液を用いることもできる。ウロン酸ユニットを含む多糖類のみを炭素源として含む培地で培養可能な細菌コロニーは、ウロン酸ユニットを含む多糖類を分解する酵素を産生していると考えられ、このような細菌コロニーの培養液のろ液は、ウロン酸ユニットを含む多糖類を分解する酵素を含むため、本実施の形態に係るバイオフィルム除去剤として用いることができる。 As the biofilm removing agent according to the present embodiment, for example, a filtrate of a culture solution obtained by culturing bacterial colonies that can be cultured in a medium containing only a polysaccharide containing a uronic acid unit as a carbon source can be used. Bacterial colonies that can be cultured in a medium containing only polysaccharides containing uronic acid units as a carbon source are considered to be producing enzymes that degrade polysaccharides containing uronic acid units. Since the filtrate contains an enzyme that decomposes a polysaccharide containing a uronic acid unit, it can be used as a biofilm remover according to the present embodiment.
本実施の形態に係るバイオフィルム除去剤は水系で水希釈物として用いるのが効果的である。本実施の形態に係るバイオフィルム除去剤で処理することにより、拭き取り、ブラッシング、水流などの物理力なしに対象物からバイオフィルムを除去することが可能であるが、短時間でのバイオフィルム除去のためには、本実施の形態に係るバイオフィルム除去剤での処理に加えて前記のような物理力を併用してもよい。除去対象のバイオフィルムが広範囲にわたる場合には、スプレー機器を用いてミストを吹き付けたり、発泡機を用いて泡状にしたものを吹き付けたりしてもよい。 It is effective to use the biofilm remover according to the present embodiment as a water dilution in an aqueous system. By treating with the biofilm remover according to the present embodiment, it is possible to remove the biofilm from the object without physical force such as wiping, brushing, water flow, etc. Therefore, in addition to the treatment with the biofilm remover according to the present embodiment, the physical force as described above may be used in combination. When the biofilm to be removed covers a wide range, mist may be sprayed using a spray device, or foamed material may be sprayed using a foaming machine.
本実施の形態に係るバイオフィルム除去剤を水系で水に溶解させて使用する場合にはバイオフィルム除去剤中の多糖類分解酵素の濃度が０．０５ｇ／Ｌ以上、好ましくは０．１ｇ／Ｌ以上、より好ましくは０．５ｇ／Ｌ以上の水溶液として使用し、バイオフィルムの処理は連続して行うことが好ましい。 When the biofilm remover according to the present embodiment is used after being dissolved in water in an aqueous system, the concentration of the polysaccharide-degrading enzyme in the biofilm remover is 0.05 g / L or more, preferably 0.1 g / L. As mentioned above, it is more preferable to use it as an aqueous solution of 0.5 g / L or more, and to carry out biofilm processing continuously.
本実施の形態に係るバイオフィルム除去剤は、バイオフィルムの危害が及ぶ広い分野に使用することができる。例えば、菌汚染リスクの高い食品製造現場、キッチン、厨房などの洗浄、医療機器の洗浄、逆浸透膜や精密ろ過膜の洗浄の際に使用することができる。また、バイオフィルムが水相中に放出される膜分離活性汚泥の場合には、本実施の形態に係るバイオフィルム除去剤で分離膜や活性汚泥を処理するよう、活性汚泥槽等へ添加して使用することができる。 The biofilm remover according to the present embodiment can be used in a wide range of fields where biofilm harms. For example, it can be used for cleaning food production sites, kitchens, kitchens, etc. at high risk of bacterial contamination, cleaning medical equipment, and cleaning reverse osmosis membranes and microfiltration membranes. In addition, in the case of membrane-separated activated sludge that is released into the aqueous phase, it is added to the activated sludge tank or the like so as to treat the separation membrane or activated sludge with the biofilm remover according to the present embodiment. Can be used.
（膜分離方法）
本実施の形態に係るバイオフィルム除去剤は、分離膜が設置された分離膜装置を用いて被処理水をろ過する工程を含む膜分離方法において使用することができる。前記分離膜を本実施の形態に係るバイオフィルム除去剤で処理することにより、膜の目詰まりの原因となるバイオフィルムを分解および除去し、膜差圧の上昇を抑制することができる。これにより、長期間安定的に膜分離方法による水処理を行うことができる。
(Membrane separation method)
The biofilm removing agent according to the present embodiment can be used in a membrane separation method including a step of filtering water to be treated using a separation membrane device in which a separation membrane is installed. By treating the separation membrane with the biofilm remover according to the present embodiment, the biofilm that causes clogging of the membrane can be decomposed and removed, and an increase in membrane differential pressure can be suppressed. Thereby, the water treatment by a membrane separation method can be performed stably for a long period of time.
（膜分離活性汚泥法）
上記の膜分離方法の一例として、本実施の形態に係るバイオフィルム除去剤を好適に用いることができる膜分離活性汚泥法について説明する。
(Membrane separation activated sludge method)
As an example of the above membrane separation method, a membrane separation activated sludge method in which the biofilm remover according to the present embodiment can be suitably used will be described.
膜分離活性汚泥法は、微生物を含む活性汚泥を収容した活性汚泥槽に有機性廃水を流入させる流入工程、および、この活性汚泥槽で生物的に処理した処理液を、活性汚泥槽に設置した分離膜装置によって固液分離する分離工程を含む。 In the membrane separation activated sludge method, an inflow process in which organic wastewater is introduced into an activated sludge tank containing activated sludge containing microorganisms, and a treatment liquid biologically treated in the activated sludge tank are installed in the activated sludge tank. Including a separation step of solid-liquid separation by a separation membrane device.
流入工程では、まず、有機性廃水から夾雑物を取り除く前処理設備によって、大きな固形分などをおおまかに除去する。前処理された有機性廃水は、流量調節槽にいったん蓄えられた後、流量を調節しながら活性汚泥槽に送り込まれる。 In the inflow process, first, large solids are roughly removed by a pretreatment facility that removes impurities from organic wastewater. The pretreated organic wastewater is once stored in the flow rate control tank, and then sent to the activated sludge tank while adjusting the flow rate.
続く分離工程では、まずこの活性汚泥槽において、活性汚泥中の微生物により有機性廃水中の有機物が分解される（生分解可能な該有機物をＢＯＤ成分とも呼ぶ）。活性汚泥槽の大きさおよび有機性廃水の活性汚泥槽での滞留時間は、活性汚泥槽における有機性廃水の処理量や、有機性廃水中の有機物濃度に応じて適宜決定することができる。活性汚泥槽中の活性汚泥濃度は一般に５〜２０ｇ／Ｌ程度が好ましいが、この範囲に限定されない。 In the subsequent separation step, first, in the activated sludge tank, organic matter in the organic wastewater is decomposed by microorganisms in the activated sludge (the biodegradable organic matter is also referred to as a BOD component). The size of the activated sludge tank and the residence time of the organic waste water in the activated sludge tank can be appropriately determined according to the treatment amount of the organic waste water in the activated sludge tank and the organic substance concentration in the organic waste water. The activated sludge concentration in the activated sludge tank is generally preferably about 5 to 20 g / L, but is not limited to this range.
分離工程では、次に、分離膜装置によって、活性汚泥槽中の活性汚泥と有機性廃水との固液分離を行う。活性汚泥槽に設置された浸漬型分離膜装置は、分離膜と集水部を含む。分離膜装置の集水部は吸引ポンプに配管され、吸引ポンプによって膜の内面と外面に圧力勾配が発生し、固液分離が達成される。この活性汚泥槽中で、活性汚泥中および有機性廃水中の微生物等によりバイオフィルムが形成され、分離膜の目詰まりの一因となる。したがって、本実施の形態に係るバイオフィルム除去剤で分離膜や活性汚泥を処理するために、活性汚泥等に本実施の形態に係るバイオフィルム除去剤を添加することにより、分離膜の目詰まりを防止し、長期間安定的に膜分離活性汚泥法による有機性廃水の処理を行うことができる。 Next, in the separation step, solid-liquid separation between the activated sludge in the activated sludge tank and the organic waste water is performed by the separation membrane device. The submerged separation membrane apparatus installed in the activated sludge tank includes a separation membrane and a water collection unit. The water collecting part of the separation membrane device is connected to a suction pump, and a pressure gradient is generated between the inner surface and the outer surface of the membrane by the suction pump to achieve solid-liquid separation. In this activated sludge tank, a biofilm is formed by microorganisms in the activated sludge and the organic waste water, which contributes to clogging of the separation membrane. Therefore, in order to treat the separation membrane and activated sludge with the biofilm remover according to this embodiment, the biofilm remover according to this embodiment is added to the activated sludge and the like, thereby clogging the separation membrane. Therefore, it is possible to stably treat organic wastewater by the membrane separation activated sludge method for a long period of time.
分離膜に用いられる膜カートリッジには、平膜、中空糸膜など公知の分離膜を用いることができる。中でも、中空糸膜は膜自身の強度が高く、有機性廃水中の夾雑物との接触から膜表面に受けるダメージが少なく、比較的長期間の使用に耐えることができる。 As the membrane cartridge used for the separation membrane, a known separation membrane such as a flat membrane or a hollow fiber membrane can be used. Among these, the hollow fiber membrane has a high strength, and is less damaged by contact with contaminants in organic wastewater, and can withstand long-term use.
分離膜の孔径や素材は、ろ過が好適に実施される限り特に限定されないが、例えば、孔径０．１μｍ程度で、ポリフッ化ビニリデン（ＰＶＤＦ）製のものが好ましく用いられる。 The pore size and material of the separation membrane are not particularly limited as long as filtration is suitably performed. For example, those having a pore size of about 0.1 μm and made of polyvinylidene fluoride (PVDF) are preferably used.
分離膜の目詰まりを防ぐためには、本実施の形態に係るバイオフィルム除去剤による処理に加え、物理的手法として、分離膜装置にスカートを設置してスカートへブロワーから気体を送り込んで膜を揺動させたり、膜面に水流をあててせん断力を与えたりしてもよい。ろ過方向とは逆方向にろ過水等を噴出させることによってバイオフィルム等膜表面の付着物を除去する逆洗を行ってもよい。 In order to prevent clogging of the separation membrane, in addition to the treatment with the biofilm remover according to the present embodiment, as a physical method, a skirt is installed in the separation membrane device and gas is sent from the blower to the skirt to shake the membrane. It may be moved, or a shearing force may be applied by applying a water flow to the membrane surface. You may perform backwashing which removes the deposit | attachment on membrane surfaces, such as a biofilm, by ejecting filtered water etc. in the reverse direction to the filtration direction.
分離膜装置は、活性汚泥槽内に浸漬して設置するだけでなく、活性汚泥槽の後段に接続して設置してもよい。したがって本実施の形態に係るバイオフィルム除去剤は、分離膜装置浸漬型の膜分離活性汚泥法だけでなく、分離膜装置を活性汚泥槽とは別の槽へ設ける場合や、加圧型の分離膜装置を用いる場合にも適用できる。これらの方法の場合は、活性汚泥槽と分離膜装置の間で活性汚泥を循環させ、濃縮液を活性汚泥槽へ戻す。また、これらの方法の場合、バイオフィルム除去剤は、分離膜装置および活性汚泥槽の両方に添加することも、どちらか一方に添加することもできる。どちらか一方に添加する場合、本実施の形態に係るバイオフィルム除去剤で分離膜を処理して分離膜の目詰まりを効率よく防止するという観点から、分離膜装置への添加が好ましい。 The separation membrane device may be installed not only by being immersed in the activated sludge tank, but also connected to the subsequent stage of the activated sludge tank. Therefore, the biofilm remover according to the present embodiment is not limited to the separation membrane device immersion type membrane separation activated sludge method, but when the separation membrane device is provided in a tank different from the activated sludge tank, or a pressure type separation membrane. The present invention can also be applied when using an apparatus. In the case of these methods, the activated sludge is circulated between the activated sludge tank and the separation membrane device, and the concentrated liquid is returned to the activated sludge tank. In these methods, the biofilm remover can be added to both the separation membrane device and the activated sludge tank, or can be added to either one. When added to either one, the addition to the separation membrane device is preferable from the viewpoint of efficiently preventing clogging of the separation membrane by treating the separation membrane with the biofilm removing agent according to the present embodiment.
分離膜は必要に応じて複数系列としてもよい。複数系列とすることにより、分離膜の系列毎に分離作業を行ったり分離作業を止めたりすることもできるので、廃水処理スピードの調整や分離膜の管理が可能になる。この場合、複数の分離膜のそれぞれを本実施の形態に係るバイオフィルム除去剤で処理することが好ましい。 A plurality of separation membranes may be used as necessary. By using a plurality of series, it is possible to perform separation work for each separation membrane series or to stop the separation work. Therefore, it is possible to adjust the wastewater treatment speed and manage the separation membrane. In this case, it is preferable to treat each of the plurality of separation membranes with the biofilm remover according to the present embodiment.
本実施の形態のバイオフィルム除去剤を用いた膜分離活性汚泥法で処理することのできる有機性廃水は、特に限定されないが、例えば、食品工場廃水、製糖工場廃水、洗剤工場廃水、スターチ工場廃水、豆腐工場廃水などが挙げられる。 The organic wastewater that can be treated by the membrane separation activated sludge method using the biofilm remover of the present embodiment is not particularly limited. For example, food factory wastewater, sugar factory wastewater, detergent factory wastewater, and starch factory wastewater. And tofu factory wastewater.
（廃水処理装置）
上記の膜分離活性汚泥法による廃水処理方法に用いる処理に用いる装置としては、例えば、図１に示される装置が挙げられる。
(Waste water treatment equipment)
As an apparatus used for the process used for the wastewater treatment method by said membrane separation activated sludge method, the apparatus shown by FIG. 1 is mentioned, for example.
まず、有機性廃水１は、前処理設備２で夾雑物を除去した後に流量調整槽３に一旦貯留され、流量調整槽３から一定の流量で活性汚泥槽（曝気槽）４に供給される。 First, the organic wastewater 1 is temporarily stored in the flow rate adjusting tank 3 after removing impurities by the pretreatment facility 2, and is supplied from the flow rate adjusting tank 3 to the activated sludge tank (aeration tank) 4 at a constant flow rate.
活性汚泥槽４では、槽に入れられた活性汚泥中の微生物によって有機性廃水１中の有機物（ＢＯＤ成分）が分解除去される。活性汚泥槽４における活性汚泥混合液の固液分離は槽内に浸漬された分離膜装置５で行う。活性汚泥槽４に本実施の形態のバイオフィルム除去剤を添加して分離膜装置５を処理することにより、バイオフィルムによる分離膜装置５の目詰まりを防止することができる。例えば、ウロン酸ユニット濃度の経時的な変化を測定する濃度測定手段としてウロン酸ユニット濃度測定装置１３を活性汚泥槽４中に設け、該濃度測定装置で測定したウロン酸ユニット濃度が所定の濃度に達したときに、バイオフィルム除去剤を活性汚泥中に添加するバイオフィルム除去剤添加装置１４を設けることにより、本実施の形態のバイオフィルム除去剤を添加することができる。また、分離膜装置５の下部にはスカート６およびブロワー７が設置されており、スカート６へブロワー７から気体が送り込まれることで、物理的にも分離膜装置５の目詰まりを防止することができる。分離膜装置５で処理されたろ過液９は、吸引ポンプ８で吸引されて、必要に応じて滅菌槽１０で消毒後、処理水１１として放流される。余剰汚泥１５は、必要に応じて活性汚泥槽（曝気槽）４から汚泥引き抜きポンプ１２により引き抜かれる。 In the activated sludge tank 4, the organic matter (BOD component) in the organic waste water 1 is decomposed and removed by microorganisms in the activated sludge put in the tank. Solid-liquid separation of the activated sludge mixed liquid in the activated sludge tank 4 is performed by the separation membrane device 5 immersed in the tank. By adding the biofilm remover of the present embodiment to the activated sludge tank 4 and treating the separation membrane device 5, clogging of the separation membrane device 5 due to biofilm can be prevented. For example, a uronic acid unit concentration measuring device 13 is provided in the activated sludge tank 4 as a concentration measuring means for measuring a change in uronic acid unit concentration with time, and the uronic acid unit concentration measured by the concentration measuring device is set to a predetermined concentration. When the biofilm remover is reached, the biofilm remover of the present embodiment can be added by providing the biofilm remover addition device 14 for adding the biofilm remover into the activated sludge. In addition, a skirt 6 and a blower 7 are installed in the lower part of the separation membrane device 5, and the clogging of the separation membrane device 5 can be physically prevented by sending gas from the blower 7 to the skirt 6. it can. The filtrate 9 treated by the separation membrane device 5 is sucked by the suction pump 8, disinfected in the sterilization tank 10 as necessary, and then discharged as treated water 11. Excess sludge 15 is extracted from the activated sludge tank (aeration tank) 4 by a sludge extraction pump 12 as necessary.
（廃水処理方法）
上記の廃水処理装置を用いた本実施の形態に係る廃水処理方法において、前記活性汚泥槽の水相中のウロン酸ユニット濃度の経時的な変化を測定し、該ウロン酸ユニット濃度が所定の濃度、例えば３０ｍｇ／Ｌに達したときに、本実施の形態に係るバイオフィルム除去剤を添加することができる。
(Waste water treatment method)
In the wastewater treatment method according to the present embodiment using the above wastewater treatment apparatus, a change with time of the uronic acid unit concentration in the aqueous phase of the activated sludge tank is measured, and the uronic acid unit concentration is a predetermined concentration. For example, when it reaches 30 mg / L, the biofilm removing agent according to the present embodiment can be added.
上述のとおり、膜分離活性汚泥法における分離膜の目詰まりは、活性汚泥中の微生物が代謝する多糖類を含むバイオフィルムが一因である。ウロン酸ユニットを含む多糖類を分解する酵素を含む、本実施の形態に係るバイオフィルム除去剤によって、バイオフィルムを除去することができるため、活性汚泥槽に収容された活性汚泥の水相中の糖濃度、好ましくはウロン酸ユニット濃度を測定し、これに基づいて本実施の形態に係るバイオフィルム除去剤を適時に適量加えて、安定した膜分離を効率よく達成することができる。 As described above, the clogging of the separation membrane in the membrane separation activated sludge method is partly due to a biofilm containing polysaccharides metabolized by microorganisms in the activated sludge. Since the biofilm can be removed by the biofilm removing agent according to the present embodiment, which includes an enzyme that decomposes the polysaccharide containing the uronic acid unit, the water in the activated sludge contained in the activated sludge tank A stable membrane separation can be efficiently achieved by measuring a sugar concentration, preferably a uronic acid unit concentration, and adding an appropriate amount of the biofilm removing agent according to the present embodiment on a timely basis.
一般に、活性汚泥の水相中のウロン酸ユニット濃度は、好ましくは５０ｍｇ／Ｌ以下であり、より好ましくは３０ｍｇ／Ｌ以下であり、さらに好ましくは２０ｍｇ／Ｌ以下、最も好ましくは１０ｍｇ／Ｌである。従って、本実施の形態に係るバイオフィルム除去剤は、このウロン酸ユニット濃度をこれらの濃度以下に維持するように添加することが好ましいが、一般に、０．６ｍ／Ｄａｙ以下のフラックスで運転した場合には、ウロン酸ユニット濃度が３０ｍｇ／Ｌ以下において膜の目詰まりが起こりにくい傾向にあるため、ウロン酸ユニット濃度が３０ｍｇ／Ｌに達したときにバイオフィルム除去剤を添加すれば、経済的にも有利である。一方、ウロン酸ユニット濃度が５０ｍｇ／Ｌになると膜の目詰まりが起こり始めるため、５０ｍｇ／Ｌに達する前、好ましくは４０ｍｇ／Ｌに達する前に本実施の形態に係るバイオフィルム除去剤を添加することが望ましい。 In general, the uronic acid unit concentration in the aqueous phase of activated sludge is preferably 50 mg / L or less, more preferably 30 mg / L or less, even more preferably 20 mg / L or less, and most preferably 10 mg / L. . Therefore, the biofilm remover according to the present embodiment is preferably added so that the uronic acid unit concentration is maintained below these concentrations, but generally when operated at a flux of 0.6 m / day or less. Since there is a tendency that clogging of the membrane does not easily occur when the uronic acid unit concentration is 30 mg / L or less, it is economically possible to add a biofilm remover when the uronic acid unit concentration reaches 30 mg / L. Is also advantageous. On the other hand, when the uronic acid unit concentration reaches 50 mg / L, clogging of the film begins to occur. Therefore, the biofilm remover according to the present embodiment is added before reaching 50 mg / L, preferably before reaching 40 mg / L. It is desirable.
活性汚泥の水相中のウロン酸ユニット濃度を測定するためには、ろ紙など、分離膜装置の分離膜より大きな孔径を有するろ材によってろ過して汚泥ろ液を得てから、測定することが好ましい。この操作によって、活性汚泥中の浮遊物のみがろ材に捕捉され、糖成分はろ紙を通過する。したがってそのろ液中の全糖ユニット濃度および／またはウロン酸ユニット濃度を測定すれば膜の目詰まり物質（バイオフィルム）となる生物由来多糖類の濃度をより正確に知ることができる。 In order to measure the uronic acid unit concentration in the aqueous phase of activated sludge, it is preferable to measure after obtaining a sludge filtrate by filtering with a filter medium having a larger pore size than the separation membrane of the separation membrane device, such as filter paper. . By this operation, only the suspended matter in the activated sludge is captured by the filter medium, and the sugar component passes through the filter paper. Therefore, by measuring the total sugar unit concentration and / or the uronic acid unit concentration in the filtrate, the concentration of the biological polysaccharide that becomes a membrane clogging substance (biofilm) can be known more accurately.
ろ材の孔径は、分離膜装置に備えられた分離膜の孔径の好ましくは５倍以上、さらに好ましくは１０倍以上である。また、分離膜装置に備えられた分離膜の約１００倍以下を上限とすることが好ましく、孔径の上限は１０μｍであることがより好ましい。さらに、親水性の素材の方が糖成分の吸着が少ないので好適であり、例えば、セルロースを素材とするろ紙を用いることができる。 The pore size of the filter medium is preferably 5 times or more, more preferably 10 times or more the pore size of the separation membrane provided in the separation membrane device. The upper limit is preferably about 100 times or less the separation membrane provided in the separation membrane device, and the upper limit of the pore diameter is more preferably 10 μm. Furthermore, a hydrophilic material is preferable because it has less adsorption of sugar components, and for example, a filter paper made of cellulose can be used.
尚、本実施の形態においてウロン酸ユニット濃度は、ウロン酸ユニット含有多糖類のウロン酸ユニットのみの濃度であり、ウロン酸濃度としてＮＥＬＬＹ ＢＬＵＭＥＮＫＲＡＮＴＺ，ＧＵＳＴＡＶ ＡＳＢＯＥ−ＨＡＮＳＥＮ著「Ｎｅｗ Ｍｅｔｈｏｄ ｆｏｒ Ｑｕａｎｔｉｔａｔｉｖｅ Ｄｅｔｅｒｍｉｎａｔｉｏｎ ｏｆ Ｕｒｏｎｉｃ Ａｃｉｄ」ＡＮＡＬＹＴＩＣＡＬ ＢＩＯＣＨＥＭＩＳＴＲＹ５４巻、４８４〜４８９貢（１９７３年発行）に記載された以下の方法に従い、ポリウロン酸の一つであるポリガラクツロン酸を用いて作成した検量線により測定することができる。
１）０．５ｍＬの汚泥ろ液および既知濃度のポリガラクツロン酸水溶液を試験管にとり、各々に３．０ｍＬの０．０１２５ＭのＮａ2Ｂ4Ｏ7濃硫酸溶液を加える。
２）各液をよくふって、沸騰湯浴中で５分間温め、その後氷水中で２０分間冷やす。
３）各液に５０μＬの０．１５％ｍ−ヒドロキシジフェニルの０．５％ＮａＯＨ溶液を加える。
４）各液をよく撹拌して、５分たったら３）の各液の５２０ｎｍ吸光度を測定し、既知濃度のポリガラクツロン酸水溶液の値と汚泥ろ液の値とを比較して濃度を求める。
In the present embodiment, the uronic acid unit concentration is a concentration of only the uronic acid unit of the uronic acid unit-containing polysaccharide, and the uronic acid concentration is NELLY BLUMENKRANTZ, GUSTAV ASBOE-HANSEN, “New Method for Quantitative Active According to the following method described in ANALYTICAL BIOCHEMISTRY Vol. 54, 484-489 Mitsugu (published in 1973), it can be measured by a calibration curve prepared using polygalacturonic acid which is one of polyuronic acids.
1) Take 0.5 mL of sludge filtrate and a known concentration of polygalacturonic acid aqueous solution into a test tube, and add 3.0 mL of 0.0125 M Na 2 B 4 O 7 concentrated sulfuric acid solution to each.
2) Wipe each solution well and warm in boiling water bath for 5 minutes, then cool in ice water for 20 minutes.
3) Add 50 μL of 0.15% m-hydroxydiphenyl in 0.5% NaOH to each solution.
4) Stir each solution well, and after 5 minutes, measure the 520 nm absorbance of each solution in 3), and compare the value of the polygalacturonic acid aqueous solution of known concentration with the value of the sludge filtrate to determine the concentration.
本実施の形態は、一態様において、膜分離活性汚泥法による有機性廃水処理装置（既存の装置であってもよい）に、活性汚泥槽の水相中のウロン酸ユニット濃度の経時的な変化を測定する濃度測定手段および該ウロン酸ユニット濃度が所定の濃度に達したときに、前記バイオフィルム除去剤を添加するバイオフィルム除去剤添加手段を備えた、膜分離活性汚泥法による有機性廃水の処理装置を提供する。該装置によれば、ウロン酸ユニット濃度に応じたバイオフィルム除去剤の添加を行うため、連続的な廃水処理を経済的に行うことができる。ウロン酸ユニット濃度の測定は、例えば上記の記載を参照して行うことができる。また、バイオフィルム除去剤添加手段は、当業者に公知に手法を用いて行うことができ、該バイオフィルム除去剤の添加を自動で行うこともできる。 In this embodiment, in one aspect, an organic wastewater treatment apparatus (which may be an existing apparatus) using a membrane separation activated sludge method is used to change the uronic acid unit concentration in the aqueous phase of the activated sludge tank over time. Organic wastewater by the membrane separation activated sludge method, comprising a concentration measuring means for measuring uronic acid units and a biofilm remover adding means for adding the biofilm remover when the uronic acid unit concentration reaches a predetermined concentration A processing device is provided. According to this apparatus, since the biofilm removing agent is added according to the uronic acid unit concentration, continuous wastewater treatment can be economically performed. The uronic acid unit concentration can be measured with reference to the above description, for example. The biofilm remover addition means can be performed using a method known to those skilled in the art, and the addition of the biofilm remover can also be performed automatically.
以下に実施例および比較例を挙げて、本実施の形態をさらに具体的に説明するが、本発明の範囲は、これらの実施例のみに限定されるものではない。
［実施例１］
リン分０．１５％、窒素分０．０４％およびＭｇ分０．００１％を含む寒天平板培地を作成した。その後、ジェランガム（シグマアルドリッチ社製）を寒天溶液に分散させ、それを前記の培地の上に積層させた。こうしてジェランガムを唯一の炭素源とする寒天平板培地を作製した。
Hereinafter, the present embodiment will be described more specifically with reference to examples and comparative examples. However, the scope of the present invention is not limited only to these examples.
[Example 1]
An agar plate medium containing 0.15% phosphorus, 0.04% nitrogen and 0.001% Mg was prepared. Thereafter, gellan gum (manufactured by Sigma-Aldrich) was dispersed in an agar solution and laminated on the medium. Thus, an agar plate medium using gellan gum as the sole carbon source was prepared.
この培地に活性汚泥、土壌および湖沼を一様に塗沫接種し、２８℃で７２時間インキュベートすると寒天平板上にカビ類、細菌類のコロニーが発生した。 When this medium was uniformly inoculated with activated sludge, soil and lakes and incubated at 28 ° C. for 72 hours, mold and bacterial colonies were formed on the agar plate.
栄養塩類としてリン分０．１５％、窒素分０．０４％およびＭｇ分０．００１％さらに、濃度１ｇ／Ｌのジェランガムを唯一の炭素源とする液体培地２０ｍＬ中に、寒天平板上に発生したカビ、細菌コロニーを白金耳で無菌的に植菌した。 As nutrients, phosphorus content was 0.15%, nitrogen content was 0.04% and Mg content was 0.001%. Furthermore, it was generated on agar plates in 20 mL of liquid medium containing gellan gum at a concentration of 1 g / L as the sole carbon source. Molds and bacterial colonies were aseptically inoculated with platinum ears.
２８℃に維持して１２０ｒｐｍで２４時間振とう通気しながら培養した。２４時間後に上述の方法でウロン酸ユニット濃度を測定したところ、ジェランガム濃度に換算すると、０．７ｇ／Ｌの濃度まで減少しており、ジェランガム分解酵素が含まれていることを確認した。以下、このジェランガム分解酵素が含まれた培養液を培養液Ａ（添加剤）と呼ぶ。 The culture was carried out with aeration at 120 rpm for 24 hours while maintaining at 28 ° C. After 24 hours, when the uronic acid unit concentration was measured by the above-described method, it was reduced to a concentration of 0.7 g / L in terms of gellan gum concentration, and it was confirmed that gellan gum-degrading enzyme was contained. Hereinafter, the culture solution containing the gellan gum-degrading enzyme is referred to as culture solution A (additive).
続いて、上述の方法で測定したウロン酸ユニット濃度が１００ｍｇ／Ｌに達した活性汚泥をアドバンテック社製５Ｃのろ紙でろ過した液１Ｌを、５Ｌ三角フラスコ中に用意し、前述の培養液Ａを１０ｍＬ、無菌的に加え、２８℃に維持して１２０ｒｐｍの速度で振とう通気処理を２４時間行った。 Subsequently, 1 L of liquid obtained by filtering the activated sludge whose uronic acid unit concentration measured by the above method reached 100 mg / L with 5C filter paper manufactured by Advantech Co., Ltd. was prepared in a 5 L Erlenmeyer flask. 10 mL, aseptically added, was maintained at 28 ° C. and aerated with shaking at a speed of 120 rpm for 24 hours.
２４時間後に上述の方法でウロン酸ユニット濃度を測定すると、５０ｍｇ／Ｌまで減少していることを確認し、その液の分子量分布を以下の条件を用いて測定すると１００万〜１億の分子量を示すピークが減少していることも確認した。 When the uronic acid unit concentration was measured by the above-mentioned method after 24 hours, it was confirmed that the concentration was reduced to 50 mg / L, and the molecular weight distribution of the liquid was measured using the following conditions. It was also confirmed that the peak shown was decreasing.
条件：分子量分布の測定にはＨＩＴＡＣＨＩ製ＨＰＬＣシステムＤ７０００シリーズを使用し、カラムはサイズ排除カラム（昭和電工製Ｓｈｏｄｅｘ−ＯＨ ｐａｃｋ ＳＢ−８０６ＨＱ）を用いた。カラム温度は４０℃、移動相には０．０５Ｍ ＫＨ2ＰＯ4 ｂｕｆｆｅｒ（ｐＨ３．０）を１．０ｍＬ／ｍｉｎの流量で送液し、２００μＬのサンプルを注入し、屈折率計により検出した。また、１×１０4、１×１０5、１×１０6、１×１０7、１×１０8および１×１０9Ｄａの分子量を有するプルランで校正曲線を描き、該校正曲線を用いてサンプルの分子量を決定した。 Conditions: A HITACHI HPLC system D7000 series was used for measuring the molecular weight distribution, and a size exclusion column (Showex-OH pack SB pack SB-806HQ) was used as the column. The column temperature was 40 ° C., 0.05M KH 2 PO 4 buffer (pH 3.0) was fed to the mobile phase at a flow rate of 1.0 mL / min, a 200 μL sample was injected, and detection was performed with a refractometer. A calibration curve is drawn with a pullulan having molecular weights of 1 × 10 4 , 1 × 10 5 , 1 × 10 6 , 1 × 10 7 , 1 × 10 8 and 1 × 10 9 Da, and a sample is obtained using the calibration curve. The molecular weight of was determined.
［実施例２］
リン分０．１５％、窒素分０．０４％、Ｍｇ分０．００１％およびキサンタンガム（シグマアルドリッチ社製）を１ｇ／Ｌを含む寒天平板培地を作成した。こうしてキサンタンガムを唯一の炭素源とする寒天平板培地を作製した。
[Example 2]
An agar plate medium containing 0.1 g of phosphorus content, 0.04% nitrogen content, 0.001% Mg content and 1 g / L of xanthan gum (manufactured by Sigma-Aldrich) was prepared. Thus, an agar plate medium using xanthan gum as the sole carbon source was prepared.
この培地に活性汚泥、土壌および湖沼を一様に塗沫接種し、２８℃で７２時間インキュベートすると寒天平板上にカビ類、細菌類のコロニーが発生した。 When this medium was uniformly inoculated with activated sludge, soil and lakes and incubated at 28 ° C. for 72 hours, mold and bacterial colonies were formed on the agar plate.
栄養塩類としてリン分０．１５％、窒素分０．０４％およびＭｇ分０．００１％さらに、濃度１ｇ／Ｌのキサンタンガムを唯一の炭素源とする液体培地２０ｍＬ中に、寒天平板上に発生したカビ、細菌コロニーを白金耳で無菌的に植菌した。 As nutrients, phosphorus content was 0.15%, nitrogen content was 0.04% and Mg content was 0.001%. Furthermore, it was generated on an agar plate in 20 mL of liquid medium containing xanthan gum as a sole carbon source at a concentration of 1 g / L. Molds and bacterial colonies were aseptically inoculated with platinum ears.
２８℃に維持して１２０ｒｐｍで２４時間振とう通気しながら培養した。２４時間後に上述の方法でウロン酸ユニット濃度を測定したところ、キサンタンガム濃度に換算すると０．６ｇ／Ｌの濃度まで減少しており、キサンタンガム分解酵素が含まれていることを確認した。以下、このキサンタンガム分解酵素が含まれた培養液を培養液Ｂ（添加剤）と呼ぶ。 The culture was carried out with aeration at 120 rpm for 24 hours while maintaining at 28 ° C. When the uronic acid unit concentration was measured by the above-mentioned method after 24 hours, it was reduced to a concentration of 0.6 g / L in terms of xanthan gum concentration, and it was confirmed that xanthan gum degrading enzyme was contained. Hereinafter, this culture solution containing xanthan gum degrading enzyme is referred to as culture solution B (additive).
続いて、上述の方法で測定したウロン酸ユニット濃度が１００ｍｇ／Ｌに達した活性汚泥をアドバンテック社製５Ｃのろ紙でろ過した液１Ｌを、５Ｌ三角フラスコ中に用意し、前述の培養液Ｂを１０ｍＬ、無菌的に加え、２８℃に維持して１２０ｒｐｍの速度で振とう通気処理を２４時間行った。 Subsequently, 1 L of liquid obtained by filtering the activated sludge whose uronic acid unit concentration measured by the above method reached 100 mg / L with 5C filter paper manufactured by Advantech Co., Ltd. was prepared in a 5 L Erlenmeyer flask. 10 mL, aseptically added, was maintained at 28 ° C. and aerated with shaking at a speed of 120 rpm for 24 hours.
２４時間後に上述の方法でウロン酸ユニット濃度を測定すると、４０ｍｇ／Ｌまで減少していることを確認し、またその液の分子量分布を実施例１と同様の手法で測定すると１００万〜１億の分子量を示すピークが減少していることも確認した。 When the uronic acid unit concentration was measured by the above-mentioned method after 24 hours, it was confirmed that the concentration was reduced to 40 mg / L, and when the molecular weight distribution of the liquid was measured by the same method as in Example 1, 1 million to 100 million It was also confirmed that the peak indicating the molecular weight of was decreased.
［実施例３］
リン分０．１５％、窒素分０．０４％、Ｍｇ分０．００１％およびヒアルロン酸１ｇ／Ｌ（シグマアルドリッチ社製）を含む寒天平板培地を作成した。こうしてヒアルロン酸を唯一の炭素源とする寒天平板培地を作製した。
[Example 3]
An agar plate medium containing 0.15% phosphorus, 0.04% nitrogen, 0.001% Mg and 1 g / L hyaluronic acid (Sigma Aldrich) was prepared. Thus, an agar plate medium using hyaluronic acid as the only carbon source was prepared.
この培地に活性汚泥、土壌および湖沼を一様に塗沫接種し、２８℃で７２時間インキュベートすると寒天平板上にカビ類、細菌類のコロニーが発生した。 When this medium was uniformly inoculated with activated sludge, soil and lakes and incubated at 28 ° C. for 72 hours, mold and bacterial colonies were formed on the agar plate.
栄養塩類としてリン分０．１５％、窒素分０．０４％およびＭｇ分０．００１％さらに、濃度１ｇ／Ｌのヒアルロン酸を唯一の炭素源とする液体培地２０ｍＬ中に、寒天平板上に発生したカビ、細菌コロニーを白金耳で無菌的に植菌した。 As nutrients, phosphorus content 0.15%, nitrogen content 0.04% and Mg content 0.001%. Furthermore, it is generated on agar plates in 20 mL of liquid medium containing only 1 g / L hyaluronic acid as a carbon source. The fungi and bacterial colonies were aseptically inoculated with platinum ears.
２８℃に維持して１２０ｒｐｍで２４時間振とう通気しながら培養した。２４時間後にウロン酸ユニット濃度を測定したところ、ヒアルロン酸濃度に換算すると０．６ｇ／Ｌの濃度まで減少しており、ヒアルロン酸分解酵素が含まれていることを確認した。以下、このヒアルロン酸分解酵素が含まれた培養液を培養液Ｃ（添加剤）と呼ぶ。 The culture was carried out with aeration at 120 rpm for 24 hours while maintaining at 28 ° C. When the uronic acid unit concentration was measured 24 hours later, it was reduced to a concentration of 0.6 g / L in terms of hyaluronic acid concentration, and it was confirmed that hyaluronic acid-degrading enzyme was contained. Hereinafter, the culture solution containing the hyaluronic acid-degrading enzyme is referred to as culture solution C (additive).
続いて、上述の方法で測定したウロン酸ユニット濃度が１００ｍｇ／Ｌに達した活性汚泥をアドバンテック社製５Ｃのろ紙でろ過した液１Ｌを、５Ｌ三角フラスコ中に用意し、前述の培養液Ｃを１０ｍＬ、無菌的に加え、２８℃に維持して１２０ｒｐｍの速度で振とう通気処理を２４時間行った。 Subsequently, 1 L of liquid obtained by filtering the activated sludge whose uronic acid unit concentration measured by the above method reached 100 mg / L with 5C filter paper manufactured by Advantech Co., Ltd. was prepared in a 5 L Erlenmeyer flask. 10 mL, aseptically added, was maintained at 28 ° C. and aerated with shaking at a speed of 120 rpm for 24 hours.
２４時間後に上述の方法でウロン酸ユニット濃度を測定すると、４９ｍｇ／Ｌまで減少していることを確認し、またその液の分子量分布を実施例１の条件と同様の手法で測定すると１００万〜１億の分子量を示すピークが減少していることも確認した。 When the uronic acid unit concentration was measured by the above method after 24 hours, it was confirmed that the uronic acid unit concentration was reduced to 49 mg / L, and the molecular weight distribution of the liquid was measured by the same method as in Example 1 to 1 million to It was also confirmed that the peak indicating a molecular weight of 100 million decreased.
［実施例４］
リン分０．１５％、窒素分０．０４％、Ｍｇ分０．００１％およびポリガラクツロン酸（シグマアルドリッチ社製）を１ｇ／Ｌを含む寒天平板培地を作成した。こうしてポリガラクツロン酸を唯一の炭素源とする寒天平板培地を作製した。
[Example 4]
An agar plate medium containing 0.1 g of phosphorus content, 0.04% nitrogen content, 0.001% Mg content and 1 g / L of polygalacturonic acid (manufactured by Sigma-Aldrich) was prepared. Thus, an agar plate medium using polygalacturonic acid as the sole carbon source was prepared.
この培地に活性汚泥、土壌および湖沼を一様に塗沫接種し、２８℃で７２時間インキュベートすると寒天平板上にカビ類、細菌類のコロニーが発生した。 When this medium was uniformly inoculated with activated sludge, soil and lakes and incubated at 28 ° C. for 72 hours, mold and bacterial colonies were formed on the agar plate.
栄養塩類としてリン分０．１５％、窒素分０．０４％およびＭｇ分０．００１％さらに、濃度１ｇ／Ｌのポリガラクツロン酸を唯一の炭素源とする液体培地２０ｍＬ中に、寒天平板上に発生したカビ、細菌コロニーを白金耳で無菌的に植菌した。 As nutrients, phosphorus content 0.15%, nitrogen content 0.04% and Mg content 0.001%. Furthermore, in 20 mL of liquid medium containing polygalacturonic acid with a concentration of 1 g / L as the sole carbon source, on an agar plate. The generated mold and bacterial colonies were aseptically inoculated with platinum ears.
２８℃に維持して１２０ｒｐｍで２４時間振とう通気しながら培養した。２４時間後に上述の方法でウロン酸ユニット濃度を測定したところ、ポリガラクツロン酸濃度に換算すると０．７ｇ／Ｌの濃度まで減少しており、ポリガラクツロン酸分解酵素が含まれていることを確認した。以下、このポリガラクツロン酸分解酵素が含まれた培養液を培養液Ｄ（添加剤）と呼ぶ。 The culture was carried out with aeration at 120 rpm for 24 hours while maintaining at 28 ° C. After 24 hours, when the uronic acid unit concentration was measured by the above-mentioned method, it was reduced to a concentration of 0.7 g / L in terms of polygalacturonic acid concentration, and it was confirmed that polygalacturonic acid-degrading enzyme was contained. . Hereinafter, the culture solution containing the polygalacturonic acid-degrading enzyme is referred to as culture solution D (additive).
続いて、上述の方法で測定したウロン酸ユニット濃度が１００ｍｇ／Ｌに達した活性汚泥をアドバンテック社製５Ｃのろ紙でろ過した液１Ｌを、５Ｌ三角フラスコ中に用意し、前述の培養液Ｄを１０ｍＬ、無菌的に加え、２８℃に維持して１２０ｒｐｍの速度で振とう通気処理を２４時間行った。 Subsequently, 1 L of liquid obtained by filtering activated sludge whose uronic acid unit concentration measured by the above method reached 100 mg / L with 5C filter paper manufactured by Advantech Co., Ltd. was prepared in a 5 L Erlenmeyer flask. 10 mL, aseptically added, was maintained at 28 ° C. and aerated with shaking at a speed of 120 rpm for 24 hours.
２４時間後に上述の方法でウロン酸ユニット濃度を測定すると、５０ｍｇ／Ｌまで減少していることを確認し、またその液の分子量分布を実施例１と同様の手法で測定すると１００万〜１億の分子量を示すピークが減少していることも確認した。 When the uronic acid unit concentration was measured by the above-mentioned method after 24 hours, it was confirmed that the concentration was reduced to 50 mg / L, and when the molecular weight distribution of the liquid was measured by the same method as in Example 1, 1 million to 100 million It was also confirmed that the peak indicating the molecular weight of was decreased.
［実施例５］
図１の系を用い、ＢＯＤが７５０ｍｇ／Ｌの製糖工場の廃水を膜分離活性汚泥法により処理した。この廃水中のウロン酸ユニット濃度は０ｍｇ／Ｌであった。
[Example 5]
Using the system shown in FIG. 1, wastewater from a sugar factory having a BOD of 750 mg / L was treated by a membrane separation activated sludge method. The uronic acid unit concentration in the wastewater was 0 mg / L.
分離膜装置５として、孔径０．１μｍの精密ろ過中空糸膜をモジュール化した分離膜装置（旭化成ケミカルズ社製、ポリフッ化ビニリデン（ＰＶＤＦ）製、膜面積０．０１５ｍ2、有効膜長さ１５ｃｍ、内径／外径：０．６／１．２ｍｍ）を、有効容積１０Ｌの活性汚泥槽４に浸漬し、吸引ろ過をした。 As the separation membrane device 5, a separation membrane device obtained by modularizing a microfiltration hollow fiber membrane having a pore diameter of 0.1 μm (manufactured by Asahi Kasei Chemicals Co., Ltd., made of polyvinylidene fluoride (PVDF), membrane area 0.015 m 2 , effective membrane length 15 cm, (Inner diameter / outer diameter: 0.6 / 1.2 mm) was immersed in an activated sludge tank 4 having an effective volume of 10 L and subjected to suction filtration.
活性汚泥槽中のＭＬＳＳ濃度は１０ｇ／Ｌで一定とし、活性汚泥槽における廃水の滞留時間は１８時間とした。処理開始時のろ過圧力は４ｋＰａであった。活性汚泥の液量は常に一定とし、ろ過Ｆｌｕｘは０．６ｍ／Ｄａｙに設定してろ液は全量活性汚泥槽外に排出した。膜への曝気は、空気を膜モジュールの下部から２００Ｌ／ｈの流量で送気した。こうした装置を装置Ａ、Ｂ、Ｃ、ＤおよびＥの５つ用意し、同条件で運転を開始した。 The MLSS concentration in the activated sludge tank was fixed at 10 g / L, and the residence time of the wastewater in the activated sludge tank was 18 hours. The filtration pressure at the start of the treatment was 4 kPa. The amount of activated sludge liquid was always constant, the filtration flux was set to 0.6 m / day, and the entire amount of filtrate was discharged out of the activated sludge tank. For aeration of the membrane, air was fed from the lower part of the membrane module at a flow rate of 200 L / h. Five such devices A, B, C, D and E were prepared, and the operation was started under the same conditions.
ウロン酸ユニット濃度は上述したのと同様の手順で、グルクロン酸の検量線により求めた。１日に１回、活性汚泥の水相中のウロン酸ユニット濃度を測定した。 The uronic acid unit concentration was determined by a glucuronic acid calibration curve in the same procedure as described above. Once a day, the uronic acid unit concentration in the aqueous phase of activated sludge was measured.
運転開始から約１週間を経過すると活性汚泥の水相中のウロン酸ユニット濃度が急激に上昇し、１１日目の装置Ａ、Ｂ、Ｃ、ＤおよびＥの活性汚泥の水相中のウロン酸ユニット濃度は、それぞれ５０ｍｇ／Ｌ、５５ｍｇ／Ｌ、５３ｍｇ／Ｌ、５０ｍｇ／Ｌおよび５０ｍｇ／Ｌとなった。そこで、実施例１〜４で得られた培養液Ａ、Ｂ、ＣおよびＤを、毎日１０ｍＬずつ装置Ａ、ＢおよびＣにそれぞれ加えた。運転開始から２０日目の装置Ａ、ＢおよびＣの活性汚泥の水相中のウロン酸ユニット濃度は、それぞれ２０ｍｇ／Ｌ、１５ｍｇ／Ｌ、１９ｍｇ／Ｌおよび２０ｍｇ／Ｌまで減少し、いずれの装置においても膜間差圧は急上昇することなく安定に運転することができた。 After about one week from the start of operation, the uronic acid unit concentration in the aqueous phase of activated sludge increases rapidly, and uronic acid in the aqueous phase of activated sludge of apparatuses A, B, C, D and E on the 11th day The unit concentrations were 50 mg / L, 55 mg / L, 53 mg / L, 50 mg / L and 50 mg / L, respectively. Therefore, 10 mL of the culture solutions A, B, C, and D obtained in Examples 1 to 4 were added to the devices A, B, and C each day. The concentration of uronic acid units in the aqueous phase of the activated sludge of devices A, B and C on the 20th day from the start of operation decreased to 20 mg / L, 15 mg / L, 19 mg / L and 20 mg / L, respectively. However, the transmembrane pressure difference could be stably operated without increasing rapidly.
一方、装置Ｅでは、処理開始後２０日を経過すると、ウロン酸ユニット濃度が６０ｍｇ／Ｌになったがそのまま処理を続けた。するとその５日後には、ろ過圧力が２５ｋＰａを超え、分離膜の洗浄が必要となった。 On the other hand, in the apparatus E, when 20 days passed from the start of the treatment, the uronic acid unit concentration reached 60 mg / L, but the treatment was continued as it was. Then, after 5 days, the filtration pressure exceeded 25 kPa, and the separation membrane had to be washed.
［実施例６〜９、比較例１〜４］
実施例５で使用した装置Ａの運転１１日目の活性汚泥を捕捉粒子径１μｍのろ紙でろ過し、ろ液を得た。この液をバイオフィルム液と呼ぶ。このバイオフィルム液の分子量分布を実施例１と同様の手法で測定すると１００万から１億の分子量にメインピークを有していた。また、このバイオフィルムについて、以下の手法を用いて構成多糖を分析したところ、ヘキソース：５０％、デオキシ糖：２０％、アミノ糖：２０％およびウロン酸：１０％であった。
[Examples 6 to 9, Comparative Examples 1 to 4]
The activated sludge on the 11th day of operation of the apparatus A used in Example 5 was filtered with a filter paper having a trapped particle diameter of 1 μm to obtain a filtrate. This liquid is called a biofilm liquid. When the molecular weight distribution of this biofilm solution was measured by the same method as in Example 1, it had a main peak at a molecular weight of 1 million to 100 million. Moreover, when the constituent polysaccharide was analyzed about this biofilm using the following methods, it was hexose: 50%, deoxy sugar: 20%, amino sugar: 20%, and uronic acid: 10%.
バイオフィルムの構成多糖の分析：バイオフィルムサンプルは酸によって単糖まで加水分解し、ＰＭＰ（１−Ｐｈｅｎｙｌ−３−ｍｅｔｈｙｌ−５−ｐｙｒａｚｏｌｏｎｅ）でラベル化した。こうして得られたＰＭＰラベル化バイオフィルムサンプルはＬＣ／ＭＳ（アジレント製、カラム：ＣＤ−Ｃ１８、移動相流量：０．２ｍｌ／ｍｉｎ、カラム温度：４０℃）を用いて分析した。溶離液には０．１Ｍ酢酸アンモニウム緩衝液と１００％メタノールを用い、１０−１００％の範囲で濃度勾配をかけ、２５４ｎｍのＵＶ吸収により構成多糖を検出した。 Biofilm constituent polysaccharide analysis: Biofilm samples were hydrolyzed to acid monosaccharides with acid and labeled with PMP (1-Phenyl-3-methyl-5-pyrazole). The PMP-labeled biofilm sample thus obtained was analyzed using LC / MS (manufactured by Agilent, column: CD-C18, mobile phase flow rate: 0.2 ml / min, column temperature: 40 ° C.). Using 0.1M ammonium acetate buffer and 100% methanol as the eluent, a concentration gradient was applied in the range of 10-100%, and the constituent polysaccharide was detected by UV absorption at 254 nm.
表１の比較例１〜４に示す市販の酵素を各０．０５ｇ／Ｌの濃度で５０ｍＭトリス塩酸緩衝液に溶解した溶液を、上記のバイオフィルム液と１：１で混合し、２８℃で３時間インキュベート後の分子量分布を実施例１と同様の手法で測定した。 A solution prepared by dissolving the commercially available enzymes shown in Comparative Examples 1 to 4 in Table 1 in a 50 mM Tris-HCl buffer at a concentration of 0.05 g / L was mixed 1: 1 with the above biofilm solution at 28 ° C. The molecular weight distribution after incubation for 3 hours was measured in the same manner as in Example 1.
一方、実施例１〜４で得られた培養液Ａ〜Ｄについて、捕捉粒子径０．４５μｍのフィルターでろ過して微生物を除去した液を、それぞれ培養液ろ液Ａ〜Ｄとした。これらの培養液ろ液は、それぞれ表１に記載の酵素を含む。これらの培養液ろ液についてもそれぞれバイオフィルム液と１：１で混合し、２８℃で３時間インキュベート後の分子量分布を実施例１と同様の手法で測定した。 On the other hand, for the culture solutions A to D obtained in Examples 1 to 4, the solutions obtained by filtering the microorganisms by filtering with a filter having a trapped particle size of 0.45 μm were used as the culture solution filtrates A to D, respectively. Each of these culture filtrates contains the enzymes listed in Table 1. Each of these culture filtrates was mixed with the biofilm solution at a ratio of 1: 1, and the molecular weight distribution after incubation at 28 ° C. for 3 hours was measured in the same manner as in Example 1.
結果を表１に示す。表１中、分解率は、処理前のバイオフィルム液の１００万から１億の分子量帯のピーク高さをもとに、以下の式を用いて算出した。
分解率＝１００×（１−（インキュベート後のピーク高さ）／（インキュベート前のピーク高さ）
The results are shown in Table 1. In Table 1, the degradation rate was calculated using the following formula based on the peak height in the molecular weight band from 1 million to 100 million of the biofilm solution before treatment.
Degradation rate = 100 × (1− (peak height after incubation) / (peak height before incubation)
表１から、比較例１〜４に示す市販の酵素を用いた場合、バイオフィルムの分解率は低いことが明らかになった。比較例１〜４で用いた酵素はいずれも細菌の細胞壁等の成分を分解する作用を有する酵素であるが、これらの酵素の作用ではバイオフィルムを高い割合で分解することはできなかった。 From Table 1, when the commercially available enzyme shown to Comparative Examples 1-4 was used, it became clear that the decomposition rate of a biofilm is low. The enzymes used in Comparative Examples 1 to 4 are all enzymes that have an action of decomposing components such as bacterial cell walls. However, the biofilm could not be decomposed at a high rate by the action of these enzymes.
一方、培養液ろ液Ａ〜Ｄを用いた場合、いずれもバイオフィルムの分解率が高かった。この結果から、培養液ろ液Ａ〜Ｄに含まれる、ジェランガム、キサンタンガム、ヒアルロン酸またはポリガラクツロン酸を分解する酵素により、バイオフィルムを効率よく分解できると考えられた。
リン分０．１５％、窒素分０．０４％、Ｍｇ分０．００１％およびグルコース（シグマアルドリッチ社製）１ｇ／Ｌを含む寒天平板培地を作成した。こうしてグルコースを唯一の炭素源とする寒天平板培地を作製した。
On the other hand, when the culture filtrates A to D were used, the biofilm decomposition rate was high. From this result, it was considered that the biofilm can be efficiently decomposed by an enzyme that decomposes gellan gum, xanthan gum, hyaluronic acid or polygalacturonic acid contained in the culture fluid filtrates A to D.
An agar plate medium containing 0.15% phosphorus, 0.04% nitrogen, 0.001% Mg and 1 g / L glucose (Sigma Aldrich) was prepared. Thus, an agar plate medium using glucose as the sole carbon source was prepared.
この培地に活性汚泥、土壌および湖沼を一様に塗沫接種し、２８℃で７２時間インキュベートすると寒天平板上に細菌類のコロニーが発生した。 When activated sludge, soil and lake were uniformly inoculated on this medium and incubated at 28 ° C. for 72 hours, bacterial colonies were formed on the agar plate.
栄養塩類としてリン分０．１５％、窒素分０．０４％およびＭｇ分０．００１％さらに、濃度１ｇ／Ｌのグルコースを唯一の炭素源とする液体培地２０ｍＬ中に、寒天平板上に発生した細菌コロニーを白金耳で無菌的に植菌した。 As nutrients, phosphorus content was 0.15%, nitrogen content was 0.04% and Mg content was 0.001%. Furthermore, it was generated on agar plates in 20 mL of liquid medium containing glucose at a concentration of 1 g / L as the sole carbon source. Bacterial colonies were aseptically inoculated with platinum ears.
２８℃に維持して１２０ｒｐｍで２４時間振とう通気しながら培養した。２４時間後に全糖濃度を測定したところ、グルコース濃度に換算すると０．６ｇ／Ｌの濃度まで減少しており、グルコース分解酵素が含まれていることを確認した。以下、このグルコース分解酵素が含まれた培養液を培養液Ｅ（添加剤）と呼ぶ。 The culture was carried out with aeration at 120 rpm for 24 hours while maintaining at 28 ° C. When the total sugar concentration was measured after 24 hours, it was reduced to a concentration of 0.6 g / L in terms of glucose concentration, and it was confirmed that glucose-degrading enzyme was contained. Hereinafter, the culture solution containing the glucose degrading enzyme is referred to as a culture solution E (additive).
続いて、上述の方法で測定したウロン酸ユニット濃度が１００ｍｇ／Ｌに達した活性汚泥をアドバンテック社製５Ｃのろ紙でろ過した液１Ｌを、５Ｌ三角フラスコ中に用意し、前述の培養液Ｅを１０ｍＬ、無菌的に加え、２８℃に維持して１２０ｒｐｍの速度で振とう通気処理を２４時間行った。 Subsequently, 1 L of liquid obtained by filtering the activated sludge whose uronic acid unit concentration measured by the above method reached 100 mg / L with 5C filter paper manufactured by Advantech Co., Ltd. was prepared in a 5 L Erlenmeyer flask. 10 mL, aseptically added, was maintained at 28 ° C. and aerated with shaking at a speed of 120 rpm for 24 hours.
２４時間後に上述の方法でウロン酸ユニット濃度を測定すると、１００ｍｇ／Ｌとほとんど変化しておらず、またその液の分子量分布を実施例１の条件と同様の手法で測定すると１００万〜１億の分子量を示すピークも全く変化しておらず、多糖類の分解は確認されなかった。 When the uronic acid unit concentration was measured by the above-mentioned method after 24 hours, it hardly changed to 100 mg / L, and when the molecular weight distribution of the liquid was measured by the same method as in the conditions of Example 1, 1 million to 100 million The peak indicating the molecular weight was not changed at all, and degradation of the polysaccharide was not confirmed.
［比較例６］
リン分０．１５％、窒素分０．０４％、Ｍｇ分０．００１％および炭素源としてペプトンおよびポリガラクツロン酸（シグマアルドリッチ社製）をそれぞれ０．５ｇ／Ｌを含む寒天平板培地を作成した。
[Comparative Example 6]
An agar plate medium containing 0.15% phosphorus, 0.04% nitrogen, 0.001% Mg, and 0.5 g / L each of peptone and polygalacturonic acid (Sigma Aldrich) as a carbon source was prepared. .
この培地に活性汚泥、土壌および湖沼を一様に塗沫接種し、２８℃で７２時間インキュベートすると寒天平板上に細菌類のコロニーが発生した。 When activated sludge, soil and lake were uniformly inoculated on this medium and incubated at 28 ° C. for 72 hours, bacterial colonies were formed on the agar plate.
栄養塩類としてリン分０．１５％、窒素分０．０４％およびＭｇ分０．００１％さらに、ペプトンおよびポリガラクツロン酸をそれぞれ濃度０．５ｇ／Ｌ含む、液体培地２０ｍＬ中に、寒天平板上に発生した細菌コロニーを白金耳で無菌的に植菌した。 As nutrients, phosphorus content 0.15%, nitrogen content 0.04%, Mg content 0.001%, and peptone and polygalacturonic acid in a concentration of 0.5 g / L, respectively. The resulting bacterial colonies were aseptically inoculated with platinum ears.
２８℃に維持して１２０ｒｐｍで２４時間振とう通気しながら培養した。全糖濃度は０．５ｇ／Ｌまで減少していた。以下、この培養液を培養液Ｆ（添加剤）と呼ぶ。 The culture was carried out with aeration at 120 rpm for 24 hours while maintaining at 28 ° C. The total sugar concentration was reduced to 0.5 g / L. Hereinafter, this culture solution is referred to as culture solution F (additive).
続いて、上述の方法で測定したウロン酸ユニット濃度が１００ｍｇ／Ｌに達した活性汚泥をアドバンテック社製５Ｃのろ紙でろ過した液１Ｌを、５Ｌ三角フラスコ中に用意し、前述の培養液Ｆを１０ｍＬ、無菌的に加え、２８℃に維持して１２０ｒｐｍの速度で振とう通気処理を２４時間行った。 Subsequently, 1 L of liquid obtained by filtering the activated sludge whose uronic acid unit concentration measured by the above-mentioned method reached 100 mg / L with 5C filter paper manufactured by Advantech Co., Ltd. was prepared in a 5 L Erlenmeyer flask, 10 mL, aseptically added, was maintained at 28 ° C. and aerated with shaking at a speed of 120 rpm for 24 hours.
２４時間後に上述の方法でウロン酸ユニット濃度を測定すると、１００ｍｇ／Ｌとほとんど変化しておらず、またその液の分子量分布を実施例１の条件と同様の手法で測定すると１００万〜１億の分子量を示すピークも全く変化しておらず、多糖類の分解は確認されなかった。 When the uronic acid unit concentration was measured by the above-mentioned method after 24 hours, it hardly changed to 100 mg / L, and when the molecular weight distribution of the liquid was measured by the same method as in the conditions of Example 1, 1 million to 100 million The peak indicating the molecular weight was not changed at all, and degradation of the polysaccharide was not confirmed.
本発明のバイオフィルム除去剤によれば、固体表面に付着したバイオフィルムや水相中に放出されたバイオフィルムを分解除去することができる。バイオフィルムは、水道管や船底での流体の摩擦抵抗の増加や、熱交換器の熱移動の減少、金属材料の腐食や、食品、医療機器の汚染など、種々の問題を引き起こすため、本発明は、このような問題の解決に寄与するという産業上の利用可能性を有する。特に膜分離活性汚泥法において、本発明のバイオフィルム除去剤を用いることにより、分離膜の目詰まりを防ぎ、膜分離活性汚泥法による有機性廃水の処理を長期間安定的に行うことができる。 According to the biofilm removing agent of the present invention, the biofilm attached to the solid surface or the biofilm released into the aqueous phase can be decomposed and removed. Biofilms cause various problems such as increased frictional resistance of fluids in water pipes and ship bottoms, decreased heat transfer of heat exchangers, corrosion of metal materials, and contamination of food and medical equipment. Has industrial applicability to help solve such problems. In particular, in the membrane separation activated sludge method, by using the biofilm removing agent of the present invention, clogging of the separation membrane can be prevented, and the treatment of organic wastewater by the membrane separation activated sludge method can be performed stably for a long period of time.
１…有機性廃水（流入）、２…前処理設備、３…流量調整槽、４…活性汚泥槽（曝気槽）、５…分離膜装置、６…スカート、７…ブロワー、８…吸引ポンプ、９…ろ過液、１０…滅菌槽、１１…処理水（放流）、１２…汚泥引き抜きポンプ、１３…ウロン酸ユニット濃度測定装置、１４…バイオフィルム除去剤添加装置、１５…余剰汚泥 DESCRIPTION OF SYMBOLS 1 ... Organic waste water (inflow), 2 ... Pretreatment equipment, 3 ... Flow control tank, 4 ... Activated sludge tank (aeration tank), 5 ... Separation membrane apparatus, 6 ... Skirt, 7 ... Blower, 8 ... Suction pump, DESCRIPTION OF SYMBOLS 9 ... Filtrate, 10 ... Sterilization tank, 11 ... Treated water (release), 12 ... Sludge extraction pump, 13 ... Uronic acid unit concentration measuring device, 14 ... Biofilm remover addition device, 15 ... Excess sludge
Claims (6)
前記分離膜を、請求項１に記載のバイオフィルム除去剤で処理して、膜差圧の上昇を抑制する工程を含む、膜分離方法。 A membrane separation method comprising a step of filtering water to be treated using a separation membrane device in which a separation membrane is installed,
A membrane separation method comprising the step of treating the separation membrane with the biofilm removing agent according to claim 1 to suppress an increase in membrane differential pressure.
該活性汚泥槽中またはその後段に設置した、生物処理水を固液分離する分離膜装置と、
該活性汚泥槽中の水相中のウロン酸ユニット濃度の経時的な変化を測定するウロン酸ユニット濃度測定手段と、
該ウロン酸ユニット濃度が所定の濃度に達したときに、請求項１に記載のバイオフィルム除去剤で該分離膜装置を処理するために、バイオフィルム除去剤を添加するバイオフィルム除去剤添加手段と、
を含む、膜分離活性汚泥法による有機性廃水の処理装置。 An activated sludge tank containing activated sludge containing microorganisms for biologically treating organic wastewater;
A separation membrane device for solid-liquid separation of biologically treated water, installed in the activated sludge tank or in the subsequent stage;
Uronic acid unit concentration measuring means for measuring a change with time of the uronic acid unit concentration in the aqueous phase in the activated sludge tank;
When the uronic acid unit concentration reaches a predetermined concentration, to handle the separation membrane device in biofilm removal agent according to claim 1, and biofilm removal agent addition means for adding the biofilm-removing agent ,
Organic wastewater treatment equipment using membrane separation activated sludge process.
採取した細菌コロニーを培養する工程；Culturing the collected bacterial colonies;
得られた培養液のろ液を含むバイオフィルム除去剤を得る工程；A step of obtaining a biofilm remover containing the filtrate of the obtained culture solution;
を含む、バイオフィルム除去剤の製造方法。A method for producing a biofilm remover.
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| JP4925208B2 (en) * | 2007-09-12 | 2012-04-25 | 学校法人早稲田大学 | Aerobic granule formation method, water treatment method and water treatment apparatus |
| JP5728662B2 (en) * | 2009-03-27 | 2015-06-03 | 日之出産業株式会社 | Wastewater treatment of hydrophilic polymer colloid materials |
| CN102428169B (en) * | 2009-06-22 | 2014-05-07 | 住友重机械工业株式会社 | Microbial Activity Improvement Agent, Microbial Activity Improvement Method, And Biological Waste Treatment Method |
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