JP5046665B2 - Novel microorganism and drainage purification method using the same - Google Patents
Novel microorganism and drainage purification method using the same Download PDFInfo
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- JP5046665B2 JP5046665B2 JP2007017170A JP2007017170A JP5046665B2 JP 5046665 B2 JP5046665 B2 JP 5046665B2 JP 2007017170 A JP2007017170 A JP 2007017170A JP 2007017170 A JP2007017170 A JP 2007017170A JP 5046665 B2 JP5046665 B2 JP 5046665B2
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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
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Description
本発明は、優れた有機物分解能力を有する新規微生物およびこれを用いた排水浄化方法に関する。 The present invention relates to a novel microorganism having an excellent ability to decompose organic matter and a method for purifying wastewater using the same.
従来、微生物を用いて排水を浄化したり、汚泥を分解したりする方法が広く用いられている。 Conventionally, methods for purifying waste water using microorganisms or decomposing sludge have been widely used.
例えば、特許文献1には、排水中に存在する染料などの難分解性有機物を効果的に分解するために、酸素基を導入した炭素繊維材料に、バチルス・カルボニフィルス、バチルス・フィルムス、バチルス・プミルス、シュードモナスなどから選択される複数種類の菌を固定化し、固定化された微生物と難分解性有機物を含む排水とを接触させることによって、排水を浄化する方法が記載されている。 For example, in Patent Document 1, in order to effectively decompose difficult-to-decompose organic substances such as dyes present in wastewater, carbon fiber materials into which oxygen groups have been introduced include Bacillus carbonifils, Bacillus films, A method for purifying wastewater by immobilizing a plurality of types of bacteria selected from Bacillus pumilus, Pseudomonas, and the like and bringing the immobilized microorganisms into contact with wastewater containing persistent organic substances is described.
特許文献1に記載の排水浄化方法においては、複数種類の菌を使用しているが、この場合、どの菌が有効な活動を行っているか不明であるため、有機物分解能力を管理、制御することが難しく、場合によっては浄化能力が不安定になることがある。
そこで、本発明は、有機物分解能力に優れた新規微生物、また、有機物分解能力の管理、制御がしやすい排水浄化方法を提供することを目的とする。
In the waste water purification method described in Patent Document 1, a plurality of types of bacteria are used, but in this case, it is unclear which bacteria are performing an effective activity, and therefore management and control of the organic matter decomposition ability is performed. In some cases, the purification capacity may become unstable.
Therefore, an object of the present invention is to provide a novel microorganism excellent in organic matter decomposing ability, and a wastewater purification method that can easily manage and control the organic matter decomposing ability.
本発明者は、上記問題点に鑑み、種々の鋭意研究を行った結果、厨房排水や食品工場排水などから単離して純化培養した、バチルス・プミルス(Bacilluc pumilus)属に属する新規のグラム陽性有芽胞桿菌(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 に寄託して平成19年1月22日に受領された受託番号FERM P−21162を有する新規微生物)が、単独でも優れた有機物分解能力を持つことを見出し、本発明を想到するに至った。
In view of the above problems, the present inventor has conducted various intensive studies, and as a result, has isolated a novel gram-positive gram-positive belonging to the genus Bacillus pumilus, which has been isolated and purified from kitchen effluent and food factory effluent. Spore gonococci (a new microorganism having a deposit number of FERM P-21162 , deposited on the Patent Organism Depositary, National Institute of Advanced Industrial Science and Technology, and received on January 22, 2007) has excellent organic substance decomposing ability. As a result, the present invention has been conceived.
すなわち、本発明の排水浄化方法は、有機物を分解する能力を有する受託番号FERM P−21162の、バチルス・プミルスを排水に混合し、前記排水中の有機物を分解することを特徴とする。
That is, the waste water purification method of the present invention is characterized in that Bacillus pumilus having the accession number FERM P-21162 having the ability to decompose organic matter is mixed with waste water to decompose the organic matter in the waste water.
受託番号FERM P−21162のバチルス・プミルスは、油脂、タンパク質、デンプンなどの分解能力に長け、単独でも、優れた排水浄化能力を発揮することができるので、この微生物を用いれば、有機物分解能力の管理や制御がしやすく、有機物分解能力に優れた排水浄化方法とすることができる。この微生物には、病原因子(溶血毒素、DNA分解酵素、嘔吐性毒素遺伝子)が認められず、病原性がないので人畜無害である。従って、受託番号FERM P−21162の微生物を含む培養液を、厨房排水や食品工場排水などの有機物を含む排水に投入することで、環境に負荷をかけることなく排水を浄化することができる。
Bacillus pumilus with the accession number FERM P-21162 has excellent ability to decompose oils, fats, proteins, starches, etc., and can exhibit excellent drainage purification ability by itself. It is easy to manage and control and can be a wastewater purification method with excellent organic matter decomposition ability. In this microorganism, no virulence factor (hemolytic toxin, DNA-degrading enzyme, vomiting toxin gene) is recognized, and since it is not pathogenic, it is harmless to humans. Therefore, by putting the culture solution containing the microorganism of the accession number FERM P-21162 into wastewater containing organic matter such as kitchen wastewater and food factory wastewater, the wastewater can be purified without burdening the environment.
なお、受託番号FERM P−21162の、バチルス・プミルス属に属する新規微生物を投入した排水を曝気するのが望ましい。曝気することで、排水中に酸素送り続けることができ、前記バチルス・プミルスの有機物分解を促進させることができる。
In addition, it is desirable to aerate the waste water into which the new microorganism belonging to the genus Bacillus pumilus having the accession number FERM P-21162 is added. By aeration, oxygen can continue to be fed into the waste water, and the organic matter decomposition of the Bacillus pumilus can be promoted.
ここで、受託番号FERM P−21162の、バチルス・プミルス属に属する新規微生物の菌学的性質を表1に示す。
Here, Table 1 shows the mycological properties of a novel microorganism belonging to the genus Bacillus pumilus, under the accession number FERM P-21162 .
本発明によれば、厨房排水や食品工場排水などから単離して純化培養することによって得られた受託番号FERM P−21162の、バチルス・プミルス属に属する新規微生物を用いることにより、有機物分解能力の管理や制御がしやすい排水浄化方法ならびに肥料製造方法とすることができる。また、本発明の微生物には病原性がないので、環境に負荷をかけることのない排水浄化方法とすることができる。 According to the present invention, by using a novel microorganism belonging to the genus Bacillus pumilus, the accession number FERM P-21162 obtained by isolation and purification culture from kitchen wastewater or food factory wastewater, It can be set as the waste water purification method and fertilizer manufacturing method which are easy to manage and control. In addition, since the microorganism of the present invention is not pathogenic, it can be a wastewater purification method that does not place a burden on the environment.
本発明の排水浄化方法は、受託番号FERM P−21162の、バチルス・プミルス属に属する新規微生物(以下、「本菌」と称す。)の培養液を、処理槽に貯留された厨房排水や食品工場排水に投入し、排水中に含まれる有機物を分解して浄化するものである。処理槽は、常時曝気され、これにより、本菌の有機物分解をさらに促進させることができる。ここで、本実施の形態で用いる処理槽の大きさや形状はどのようなものであってもよい。
The method for purifying wastewater of the present invention comprises a culture solution of a novel microorganism belonging to the genus Bacillus pumilus (hereinafter referred to as “the present bacterium”) having a deposit number of FERM P-21162 , and kitchen wastewater and food stored in a treatment tank. It is thrown into factory wastewater and decomposes and purifies organic substances contained in wastewater. The treatment tank is constantly aerated, thereby further promoting organic matter decomposition of the bacterium. Here, the processing tank used in the present embodiment may have any size and shape.
なお、本菌の培養液は、排水の容量の1/2000〜1/100量投入する方が望ましい。投入量が1/2000より少ないと、浄化効果が激減する。また、投入量と有機物の分解力との関係は、投入量が排水の容量の1/100程度で飽和する。したがって、これ以上投入しても効果はさほど変わらず、不必要に多くの菌液を使用することになり、コスト高となる。 In addition, it is desirable to add the culture solution of the present bacterium in an amount of 1/2000 to 1/100 of the capacity of the waste water. When the input amount is less than 1/2000, the purification effect is drastically reduced. Further, the relationship between the input amount and the decomposition power of the organic matter is saturated when the input amount is about 1/100 of the capacity of the waste water. Therefore, even if it inputs more than this, an effect does not change so much, Unnecessarily many fungus liquids will be used, and cost will become high.
本実施の形態における排水浄化方法は、以下のとおりである。
まず、排水設備に曝気装置(散気管とエアーポンプ)を取り付ける。次いで、グリストラップに付着している油脂や汚泥など内容物を清掃する。なお、無害化施設などの浄化設備の場合はそのままでよい。その後、排水の容量の1/2000〜1/100程度の本菌の培養液を、毎日1回注入する。例えば、400Lのグリストラップにあっては、200mL〜4Lである。培養液の量は、排水の汚濁度に応じて加減する。培養液の注入は、飲食店のグリストラップにあっては、排水が停止している夜間が好ましい。この場合、翌朝までの十数時間のうちに、本菌が千倍近くに増殖するため、高い浄化効果が見込まれる。なお、培養液タンクにポンプとタイマーを組み合わせた自動注入装置を用いることにより、毎日定時に一定量の培養液を注入することができる。
The waste water purification method in the present embodiment is as follows.
First, an aeration apparatus (aeration pipe and air pump) is attached to the drainage facility. Next, the contents such as oil and sludge adhering to the grease strap are cleaned. In the case of a purification facility such as a detoxification facility, it can be left as it is. Thereafter, a culture solution of the bacterium of about 1/2000 to 1/100 of the drainage capacity is injected once daily. For example, in a 400L grease strap, it is 200mL-4L. The amount of the culture solution is adjusted according to the effluent pollution degree. The culture solution is preferably injected at night when drainage is stopped in the grease trap of the restaurant. In this case, since this bacterium is proliferated nearly 1000 times in the tens of hours until the next morning, a high purification effect is expected. In addition, by using an automatic injection device that combines a pump and a timer in the culture solution tank, a fixed amount of culture solution can be injected every day at a fixed time.
以下、本発明の実施の形態に係る受託番号FERM P−21162のバチルス・プミルスの単離方法ならびに浄化能について詳細に説明する。
Hereinafter, the isolation method and purification ability of Bacillus pumilus of the accession number FERM P-21162 according to the embodiment of the present invention will be described in detail.
1−1:単離方法
多くのグリストラップから汚水試料を採取し、その希釈液を油脂分解試験用培地(ツイーン20及び無機塩類からなる培地)に接種後、30℃で3日間培養した。そして、高い菌数で油脂分解菌のコロニーが得られた培地から白金線を用いて細胞を取り、これを滅菌生理食塩水中で懸濁してから肉汁寒天培地に接種し、コロニーを形成させて、純粋な細菌株を約100菌株得た。これら細菌分離株の油脂分解能、デンプン分解能、及びタンパク質分解能をテストし、これら3つのいずれにおいても強い分解能を持つ菌株を数株得た。さらに、病原因子(溶血毒素、DNA分解酵素、及び嘔吐性毒素遺伝子)に関する試験を行い、病原因子を持たない株を選抜し、本菌を得た。
1-1: Isolation method Sewage samples were collected from many grease traps, and the diluted solution was inoculated into a fat and oil degradation test medium (medium consisting of Tween 20 and inorganic salts) and then cultured at 30 ° C. for 3 days. Then, using a platinum wire from the medium in which a colony of oleolytic bacteria was obtained with a high number of bacteria, using a platinum wire, suspending this in sterile physiological saline, inoculating into a gravy agar medium, forming a colony, About 100 pure bacterial strains were obtained. These bacterial isolates were tested for fat, starch, and protein resolution, and several strains with strong resolution in all three were obtained. Further, a test on pathogenic factors (hemolytic toxin, DNA degrading enzyme, and emetic toxin gene) was performed, and a strain having no pathogenic factor was selected to obtain the bacterium.
1−2:本菌の油脂分解能測定
1−1に記載された方法で単離された本菌を、厨房排水に1/100量接種し、30℃で24〜48時間曝気した。このときの、厨房排水に含まれるn−ヘキサン抽出物(油脂濃度)の経時変化について図1に示す。なお、比較例として、本菌を接種しない厨房排水のn−ヘキサン抽出物(油脂濃度)の経時変化についても図1に示す。
1-2: Measurement of oil-and-fat resolving power of this bacterium The present bacterium isolated by the method described in 1-1 was inoculated into kitchen drainage in an amount of 1/100 and aerated at 30 ° C. for 24-48 hours. FIG. 1 shows the change over time of the n-hexane extract (oil concentration) contained in the kitchen waste water at this time. In addition, as a comparative example, the time-dependent change of the n-hexane extract (fat concentration) of the kitchen wastewater which does not inoculate this microbe is also shown in FIG.
図1に示すように、時間の経過に伴い、本菌未接種でも厨房排水中の常在菌の効果によりn−ヘキサン抽出物(油脂濃度)は減少する(図1中白丸)が、本菌の接種により、厨房排水の浄化が促進されることがわかった(図1中黒丸)。 As shown in FIG. 1, with the passage of time, the n-hexane extract (oil concentration) decreases due to the effect of the resident bacteria in the kitchen drainage even when the bacteria are not inoculated (white circles in FIG. 1). It was found that the purification of kitchen wastewater was promoted by inoculation of (indicated by black circles in FIG. 1).
1−3:本菌の厨房排水浄化能測定
1−1に記載された方法で単離された本菌を、厨房排水に1/100量接種し、30℃で24〜48時間曝気した。このときの、厨房排水の汚染度を表す指標の1つである生物化学的酸素要求量(BOD)の経時変化について図2に示す。なお、比較例として、本菌を接種しない厨房排水のn−ヘキサン抽出物(油脂濃度)の経時変化についても図2に示す。
1-3: Measurement of kitchen drainage purification ability of the present bacteria 1/100 amount of the present bacteria isolated by the method described in 1-1 was inoculated into the kitchen drainage and aerated at 30 ° C. for 24 to 48 hours. FIG. 2 shows the change over time in the biochemical oxygen demand (BOD), which is one of the indexes representing the degree of contamination of kitchen wastewater at this time. In addition, as a comparative example, the time-dependent change of the n-hexane extract (oil concentration) of the kitchen drainage which does not inoculate this microbe is also shown in FIG.
図2に示すように、時間の経過に伴い、本菌未接種でも厨房排水中の常在菌の効果によりBODは減少する(図2中白丸)が、本菌の接種により、厨房排水の浄化が促進されることがわかった(図2中黒丸)。 As shown in FIG. 2, with the passage of time, BOD decreases due to the effect of resident bacteria in the kitchen drainage even when the bacteria are not inoculated (white circle in FIG. 2). Was found to be promoted (black circle in FIG. 2).
1−4:本菌の厨房排水浄化能と増殖能との関係
1−1に記載された方法で単離された本菌を、滅菌した厨房排水に1/2000量接種し、30℃で24〜48時間曝気した(測定A)。また、非滅菌の厨房排水に測定Aと同様の量の本菌を接種し、30℃で24〜48時間曝気した(測定B)。このときの、厨房排水に含まれるn−ヘキサン抽出物(油脂濃度)の経時変化について図3に示す。
1-4: Relationship between the ability to purify the kitchen wastewater and the growth ability of the fungus Inoculate 1/2000 amount of the fungus isolated by the method described in 1-1 into a sterilized kitchen wastewater at 30 ° C for 24 hours. Aerated for ~ 48 hours (Measurement A). In addition, non-sterile kitchen wastewater was inoculated with the same amount of the bacterium as in measurement A and aerated at 30 ° C. for 24-48 hours (measurement B). FIG. 3 shows the change over time of the n-hexane extract (oil concentration) contained in the kitchen waste water at this time.
図3に示すように、測定Aでは、約1000mg/Lの油脂濃度が24時間で約75%に、48時間で約30%に減少した。このとき、図3のCで示すように、本菌の菌数は、24時間で106CFU/mLから109CFU/mLまで約1000倍も増加しており、油脂分解に伴い本菌が油脂分を栄養源として増殖したことがわかった。 As shown in FIG. 3, in measurement A, the fat concentration of about 1000 mg / L decreased to about 75% in 24 hours and to about 30% in 48 hours. At this time, as indicated by C in FIG. 3, the number of bacteria of the bacterium increased about 10 times from 10 6 CFU / mL to 10 9 CFU / mL in 24 hours. It was found that it grew using oil and fat as nutrients.
一方、図3に示すように、測定Bでは、油脂分は24時間で約35%に減少し、48時間では約15%にまで減少した。この結果より、本菌は、厨房排水中の常在菌と共同で排水浄化に当たっていることがわかった。なお、測定Aと測定Bとの差が常在菌による油脂分解量である。 On the other hand, as shown in FIG. 3, in the measurement B, the fat and oil content decreased to about 35% in 24 hours and decreased to about 15% in 48 hours. From this result, it was found that the present bacterium was used for wastewater purification in cooperation with the resident bacteria in the kitchen effluent. The difference between measurement A and measurement B is the amount of fats and oils decomposed by resident bacteria.
なお、本菌は、単独でも優れた有機物分解能力を発揮することから、家畜の糞尿に混合させることにより、環境に負荷をかけることなく家畜の糞尿の堆肥化を図ることも可能であると推測される。 In addition, since this bacterium exhibits an excellent ability to decompose organic matter even by itself, it is estimated that it can be composted with livestock manure without burdening the environment by mixing it with livestock manure. Is done.
本発明によれば、受託番号FERM P−21162の、バチルス・プミルス属に属する新規微生物は、油脂、タンパク質、デンプンなどの分解能力に長け、単独でも、優れた有機物分解能力を発揮することができるので、分解能力が高く、また有機物分解能力の管理や制御がしやすい排水浄化方法として有用である。
According to the present invention, a novel microorganism belonging to the genus Bacillus pumilus having an accession number of FERM P-21162 is excellent in the ability to decompose oils, fats, proteins, starches, etc., and can exhibit an excellent ability to decompose organic substances alone. Therefore, it is useful as a wastewater purification method that has a high decomposing ability and is easy to manage and control the organic substance decomposing ability.
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