JP2008018413A - Method for ammonia-deodorizing, novel chlorellavulgaris having ammonia-deodorizing capacity, biomass including this novel chlorellavulgaris having ammonia-deodorizing capacity, and feed additive including this biomass - Google Patents
Method for ammonia-deodorizing, novel chlorellavulgaris having ammonia-deodorizing capacity, biomass including this novel chlorellavulgaris having ammonia-deodorizing capacity, and feed additive including this biomass Download PDFInfo
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本発明は、クロレラブルガリスを用いたアンモニア脱臭法ならびにアンモニア脱臭能を有する新規なクロレラブルガリス、およびアンモニア脱臭能を有する新規なクロレラブルガリスを含むバイオマスならびに飼料添加物に関する。 The present invention relates to an ammonia deodorizing method using Chlorella vulgaris, a novel Chlorella vulgaris having ammonia deodorizing ability, a biomass containing the novel Chlorella vulgaris having ammonia deodorizing ability, and a feed additive.
現状では、家畜糞尿は一定期間曝気処理されたのち液肥などに使用されるが、依然としてアンモニア等の悪臭が残る。また化学肥料の多用によって、利用されない糞尿が漸増しており、このため環境汚染を引き起こす懸念が高まっている。脱臭技術としては硝化細菌やアンモニア酸化細菌等の脱窒細菌等を用いた方法がある。例えば、非特許文献1,2,3には、硝化細菌を使用したアンモニア脱臭技術が、また特許文献4には、脱窒細菌を用いた窒素除去、脱臭技術が紹介されている。 At present, livestock manure is used for liquid fertilizer after aeration treatment for a certain period of time, but still a bad odor such as ammonia remains. In addition, due to heavy use of chemical fertilizers, the amount of manure that is not used is gradually increasing, which raises the concern of causing environmental pollution. As a deodorizing technique, there is a method using denitrifying bacteria such as nitrifying bacteria and ammonia oxidizing bacteria. For example, Non-Patent Documents 1, 2, and 3 introduce an ammonia deodorization technique using nitrifying bacteria, and Patent Document 4 introduces a nitrogen removal and deodorization technique using denitrifying bacteria.
非特許文献1〜3に記載されているようなアンモニア酸化細菌を用いたアンモニア脱臭には、アンモニア酸化細菌の生育には酸素が必要であることから、大量の曝気が必要となる。また、非特許文献4に記載されている脱窒細菌によるアンモニアのガス化には、炭素源としてメタノールを添加する必要があり、コスト高になるという問題がある。とくに、硝化・酸化と脱窒素が必要とされる場合には、硝化・酸化や脱窒素を行うそれぞれの細菌が好気性と嫌気性であるので、直列型の処理になり同時処理が出来ない、また、処理槽の分割・曝気処理が必要になるなど、複雑な工程とならざるを得ないという問題もある。またアンモニア酸化細菌や脱窒細菌等微生物を用いた方法は機構がまた判明しない面も多く、制御が困難である。 Ammonia deodorization using ammonia-oxidizing bacteria as described in Non-Patent Documents 1 to 3 requires a large amount of aeration since oxygen is required for growth of ammonia-oxidizing bacteria. In addition, ammonia gasification by denitrifying bacteria described in Non-Patent Document 4 requires the addition of methanol as a carbon source, which increases the cost. In particular, when nitrification / oxidation and denitrification are required, each bacterium that performs nitrification / oxidation and denitrification is aerobic and anaerobic, so it becomes a series type treatment and simultaneous treatment is not possible. In addition, there is a problem that the process tank must be a complicated process such as division of the treatment tank and aeration treatment. In addition, methods using microorganisms such as ammonia-oxidizing bacteria and denitrifying bacteria have many unclear mechanisms and are difficult to control.
そこで本発明で解決しようとする点は、これまで提案されているアンモニア酸化細菌や脱窒細菌等微生物を用いた場合に指摘されている種々の問題点を克服せんとするものである。すなわち、藻類の有する高い窒素同化能に着目し、アンモニア脱臭能を有する新しい藻類を見出して優れたアンモニア脱臭技術の確立をめざすことである。またその藻類が可及的に最大の脱臭能を発揮する培養条件について新たな技術提案を行うことである。 Therefore, the point to be solved by the present invention is to overcome various problems pointed out in the case of using microorganisms such as ammonia-oxidizing bacteria and denitrifying bacteria proposed so far. That is, paying attention to the high nitrogen assimilation ability of algae, it aims to find a new algae having ammonia deodorizing ability and to establish an excellent ammonia deodorizing technique. In addition, a new technical proposal will be made on the culture conditions under which the algae exerts the maximum possible deodorizing ability.
本発明者は、土壌中から有機物を活発に分解し、窒素利用に優れた微細藻類(クロレラ)を単離し、水溶液中でのアンモニア除去能を調べたところ、クロレラブルガリスのうち、特に、本出願人が独立行政法人産業技術総合研究所に寄託して平成18年6月26日に受領した受領番号FERM AP−20942を有するクロレラブルガリス(以下、「クロレラブルガリスNTM06」と称す。)が優れた脱臭能を有していることを見出し本発明に至った。 The present inventors actively decomposed organic matter from soil, isolated microalgae (chlorella) excellent in nitrogen utilization, and examined ammonia removal ability in an aqueous solution. Chlorella Bulgaris (hereinafter referred to as “Chlorella Bulgaris NTM06”) having the receipt number FERM AP-20942 deposited on June 26, 2006 by the applicant deposited with the National Institute of Advanced Industrial Science and Technology. It has been found that it has an excellent deodorizing ability and has led to the present invention.
すなわち、本発明は、クロレラブルガリスNTM06を用いたアンモニア脱臭法である。豚糞消化液などのアンモニアを含む水溶液中にクロレラブルガリスNTM06を添加すると、その直後に、アンモニア量が速やかに減少することがわかった。 That is, the present invention is an ammonia deodorization method using Chlorella vulgaris NTM06. It was found that when Chlorella vulgaris NTM06 was added to an aqueous solution containing ammonia such as pig swine digestive juice, the amount of ammonia rapidly decreased immediately after that.
また、本発明者は、種々の鋭意研究の結果、クロレラブルガリスNTM06を、死菌よりも生菌の状態で用いた方がアンモニアの減少比が大きくなることがわかった。すなわち、本発明のアンモニア脱臭法においては、用いるクロレラブルガリスは、アンモニアを含む水溶液中で生存可能なものである方が望ましい。これは、クロレラブルガリスNTM06が、細胞表層吸着よりも細胞内への取り込みによってアンモニア脱臭を行っている可能性が高いことを示唆している。さらに、このクロレラブルガリスNTM06によるアンモニア脱臭において、水溶液中のアンモニア濃度が高ければ高いほどアンモニアの減少比が高いことがわかった。 Further, as a result of various intensive studies, the present inventor has found that the reduction ratio of ammonia is larger when Chlorella vulgaris NTM06 is used in a live state than a dead one. That is, in the ammonia deodorization method of the present invention, it is desirable that the chlorella vulgaris used be viable in an aqueous solution containing ammonia. This suggests that Chlorella vulgaris NTM06 is more likely to deodorize ammonia by cell uptake than by cell surface adsorption. Further, in ammonia deodorization by Chlorella vulgaris NTM06, it was found that the higher the ammonia concentration in the aqueous solution, the higher the ammonia reduction ratio.
また、本発明のアンモニア脱臭法で用いるクロレラブルガリスNTM06は、グルタミン酸を0.1〜1.5%添加した培地で培養したり、培養後24〜48時間、4℃にて飢餓状態に置いたりすると、さらに脱臭能が向上することがわかった。したがって、これらの方法で培養されたクロレラブルガリスNTM06を用いることにより、本発明のアンモニア脱臭法を、さらに脱臭能力の高い脱臭法とすることができる。 Further, Chlorella vulgaris NTM06 used in the ammonia deodorization method of the present invention can be cultured in a medium supplemented with 0.1 to 1.5% glutamic acid, or placed in a starved state at 4 ° C. for 24 to 48 hours after culturing. Then, it turned out that the deodorizing ability improves further. Therefore, by using Chlorella vulgaris NTM06 cultured by these methods, the ammonia deodorization method of the present invention can be made a deodorization method with higher deodorizing ability.
また、本発明のアンモニア脱臭法で用いるアンモニア脱臭能を有するクロレラブルガリスNTM06ならびにアンモニア脱臭の過程で生育したクロレラブルガリスNTM06は、バイオマスや、水産、畜産の飼料添加物として有効である。たとえば、植物に関しては、堆肥に混合させたり、動物に関しては、飼料に混合して栄養価を高めたりすることができる。 Further, Chlorella vulgaris NTM06 having ammonia deodorizing ability used in the ammonia deodorization method of the present invention and Chlorella vulgaris NTM06 grown in the process of ammonia deodorization are effective as feed additives for biomass, fisheries and livestock. For example, the plant can be mixed with compost, and the animal can be mixed with feed to increase the nutritional value.
本発明によれば、クロレラブルガリスNTM06を用いることで、優れた脱臭能力を有し、上記従来の方法で問題となっていた、大量の曝気やメタノールなどの炭素源の添加などの操作が不必要で、処理工程を簡略化することが可能なアンモニア脱臭法とすることができる。 According to the present invention, the use of Chlorella vulgaris NTM06 has excellent deodorizing ability, and operations such as addition of a large amount of aeration and addition of a carbon source such as methanol, which have been a problem in the conventional method, are not necessary. It is possible to provide an ammonia deodorization method that can simplify the treatment process.
以下、本発明の実施の形態におけるクロレラブルガリスを用いたアンモニア脱臭法について詳細に説明する。本実施の形態にかかるアンモニア脱臭法に用いるクロレラブルガリスは、受領番号FERM AP−20942のクロレラブルガリスNTM06である。以下、このクロレラブルガリスNTM06の単離方法や形態解析の結果について説明する。 Hereinafter, the ammonia deodorization method using Chlorella vulgaris in the embodiment of the present invention will be described in detail. The chlorella bulgaris used in the ammonia deodorization method according to the present embodiment is chlorella bulgaris NTM06 having a receipt number FERM AP-20942. Hereafter, the isolation method of this Chlorella vulgaris NTM06 and the result of a form analysis are demonstrated.
(実施例1)クロレラブルガリスNTM06の単離方法
自然界から土壌を採取し、その1gに滅菌生理食塩水を10ml添加し、十分攪拌した。この懸濁液50μlを、酵母培養等に汎用されるYPD培地にストリークして培養した。このYPD培地(Bacto Peptone(Difco製):20g,Yeast extract(日本製薬製):10g,glucose:20g,water 1000ml,pH7.0)は、寒天(ナカライテスク製)を1.5%になるよう添加した後、オートクレーブ滅菌し、滅菌後に約50℃付近まで冷ました後、100μg/mlになるように抗生物質クロラムフェニコール(和光純薬製)を添加し、シャーレに固化させた(YPD平板培地)ものである。(Example 1) Isolation method of Chlorella vulgaris NTM06 Soil was collected from nature, and 10 ml of sterilized physiological saline was added to 1 g of the soil and stirred sufficiently. 50 μl of this suspension was streaked and cultured in a YPD medium generally used for yeast culture or the like. This YPD medium (Bacto Peptone (Difco): 20 g, Yeast extract (Nippon Pharmaceutical): 10 g, glucose: 20 g, water 1000 ml, pH 7.0) is 1.5% agar (Nacalai Tesque). After the addition, the mixture was sterilized by autoclave, cooled to about 50 ° C. after sterilization, and then the antibiotic chloramphenicol (manufactured by Wako Pure Chemical Industries) was added to a concentration of 100 μg / ml and solidified in a petri dish (YPD plate) Medium).
暗所で30℃、24時間保温の結果、4日で、緑色のコロニーを形成する微生物を肉眼で観察できるようになった。コロニーの形状は酵母によく似ているが、濃い緑色を呈していた。YPD液体培地にて30℃で3日間振盪培養すると、培地は深い緑色を呈した。この緑色コロニーを再度平板培地に移し、シングルコロニーアイソレーションを繰り返し、単離クロレラ菌とした。単離したクロレラの18SrRNA遺伝子の塩基配列を決定し、ホモロジーを検索すると、単離クロレラ菌はクロレラブルガリスに最も近縁であった。単離微生物はその生理学的性質からクロレラブルガリスの新種と考えられ、この単離微生物をクロレラブルガリスNTM06と命名した。 As a result of incubation at 30 ° C. for 24 hours in the dark, the microorganisms forming green colonies could be observed with the naked eye in 4 days. The shape of the colony resembled that of yeast, but it was dark green. When shake culture was performed at 30 ° C. for 3 days in YPD liquid medium, the medium showed a deep green color. This green colony was again transferred to a plate medium, and single colony isolation was repeated to obtain isolated chlorella. When the nucleotide sequence of the isolated 18S rRNA gene of chlorella was determined and the homology was searched, the isolated chlorella was most closely related to Chlorella vulgaris. The isolated microorganism is considered to be a new species of Chlorella vulgaris due to its physiological properties, and this isolated microorganism was named Chlorella vulgaris NTM06.
単離したクロレラの光学顕微鏡写真を図1に、蛍光顕微鏡(励起波長:545±15nm、放出波長:610±37.5nm)を図2に示す。単離微生物を光学顕微鏡で観察すると、図1に示すように、細胞形態は球形で大きさは5−9μmであり、細菌ではないことがわかった。UV励起下、蛍光顕微鏡で観察すると図2に示すように赤い発色が見られた。葉緑体はUV励起下では赤く発色するので、分離微生物は葉緑体を持つことが考えられた。なお、YPD液体培地における生育最適温度およびpHは、それぞれ35〜40℃、7.0であった。 An optical micrograph of the isolated chlorella is shown in FIG. 1, and a fluorescence microscope (excitation wavelength: 545 ± 15 nm, emission wavelength: 610 ± 37.5 nm) is shown in FIG. When the isolated microorganism was observed with an optical microscope, as shown in FIG. 1, it was found that the cell morphology was spherical and the size was 5-9 μm, and it was not a bacterium. When observed with a fluorescence microscope under UV excitation, a red color was observed as shown in FIG. Since chloroplasts develop a red color under UV excitation, it was considered that the separated microorganisms had chloroplasts. The optimum growth temperature and pH in the YPD liquid medium were 35 to 40 ° C. and 7.0, respectively.
なお、暗所での保温は、20〜40℃、好ましくは30℃前後の温度で3〜7日程度培養するとよい。この場合の培養では光照射はあっても、なくてもかまわない。
コロニー形成を肉眼確認(約4日)の後、低温(例えば、4℃)で保管し保存菌株とする。この保存菌株は数ヶ月毎、標準的には4か月毎で新しいYPD寒天平板培地に植え替える。The incubation in the dark is about 20 to 40 ° C., preferably about 30 ° C. for about 3 to 7 days. The culture in this case may or may not be irradiated with light.
Colony formation is visually confirmed (about 4 days), and then stored at a low temperature (for example, 4 ° C.) to obtain a preserved strain. This stock is replanted on a new YPD agar plate every few months, typically every 4 months.
(実施例2)クロレラブルガリスNTM06のアンモニア脱臭能の評価
実施例1で保管した保存菌株から白金耳を用いてコロニーをかきとり、YPD液体培地に植え付け、好ましくは37℃で数日間(例えば、4日間)、震盪培養する。これを前培養菌体とする。前培養菌体を、グルタミン酸を0.1〜1.5%含むYPD培地に移し、同様の培養条件で震盪培養する。この培養液をさらに低温(例えば、4℃)で一日前後静置することによって得られるクロレラブルガリスNTM06をアンモニア脱臭に使用することができる。クロレラブルガリスNTM06は自重で沈澱しているので、培地の上澄のみを廃棄することが好ましい。(Example 2) Evaluation of ammonia deodorizing ability of Chlorella vulgaris NTM06 The colony was scraped off from the preserved strain stored in Example 1 using a platinum loop and planted in a YPD liquid medium, preferably at 37 ° C for several days (for example, 4 Incubate for days. This is designated as a pre-cultured cell. The precultured cells are transferred to a YPD medium containing 0.1 to 1.5% glutamic acid and shake-cultured under the same culture conditions. Chlorella vulgaris NTM06 obtained by allowing this culture solution to stand still for about one day at a lower temperature (for example, 4 ° C.) can be used for ammonia deodorization. Since Chlorella vulgaris NTM06 is precipitated by its own weight, it is preferable to discard only the supernatant of the medium.
単離したクロレラブルガリスNTM06は、酢酸資化能が高いので、低級脂肪酸の同化と光合成が共役していることが考えられた。そこで低級脂肪酸等を含むと思われる豚糞消化液を用いて、光照射培養を試みた。 Since the isolated Chlorella vulgaris NTM06 has high acetic acid assimilation ability, it was considered that the assimilation of lower fatty acids and photosynthesis were coupled. Then, the light irradiation culture was tried using the digestive liquid of pig excretion which seems to contain a lower fatty acid etc.
固液分離した豚糞消化液のアンモニア濃度を血中アンモニア比色定量法にて求め、アンモニア濃度が例えば、200〜300ppmになるように希釈し、また、pHが酢酸や乳酸を用いて中性からpH6.0の間となるように調整した。この豚糞消化液に、前述の条件にて培養されたクロレラブルガリスNTM06を1mlあたり108〜1010個、最終濃度が2×1010/mlとなるように添加し、緩やかに撹拌し、そのまま静置した。すると、60分でアンモニア濃度を35ppmまで減少させることができた(図3、参照。)。The ammonia concentration of the solid-liquid separated pig feces digestion solution is obtained by a blood ammonia colorimetric determination method, diluted so that the ammonia concentration becomes, for example, 200 to 300 ppm, and the pH is neutral using acetic acid or lactic acid. To pH 6.0. To this swine manure digestive juice, Chlorella vulgaris NTM06 cultured under the above-mentioned conditions was added at 10 8 to 10 10 per ml, and the final concentration was 2 × 10 10 / ml, and gently stirred. It was left as it was. Then, in 60 minutes, the ammonia concentration could be reduced to 35 ppm (see FIG. 3).
また、アンモニア濃度が50ppmの豚糞消化液に、クロレラブルガリスNTM06を最終濃度が2×1010/mlとなるように添加すると、36時間でアンモニア濃度が0になりアンモニア臭を完全に取り除くことができた(図4、参照)。In addition, when Chlorella vulgaris NTM06 is added to the swine manure digestion solution with an ammonia concentration of 50 ppm so that the final concentration becomes 2 × 10 10 / ml, the ammonia concentration becomes 0 in 36 hours and the ammonia odor is completely removed. (See FIG. 4).
(実施例3)(グルタミン酸を添加した培地で培養したクロレラブルガリスNTM06アンモニア脱臭能の評価)
実施例1で保管した保存菌株から白金耳を用いてコロニーをかきとり、グルタミン酸0.1〜0.5%を含むYPD培地で培養し、これをアンモニア脱臭に用いた。グルタミン酸を添加したYPD培地で培養されたものは、YPD培地のみで培養されたものよりアンモニア吸収速度が20〜30%増加することがわかった。(Example 3) (Evaluation of Chlorella vulgaris NTM06 ammonia deodorizing ability cultured in a medium supplemented with glutamic acid)
Colonies were scraped from the stocks stored in Example 1 using platinum loops and cultured in YPD medium containing 0.1 to 0.5% glutamic acid, which was used for ammonia deodorization. It was found that those cultured in YPD medium supplemented with glutamic acid had a 20-30% increase in ammonia absorption rate over those cultured in YPD medium alone.
(実施例4)クロレラブルガリスNTM06の培地
実施例1におけるクロレラブルガリスNTM06は、YPD培地で最もよく生育した。そこで、グルコース濃度を変えて得られる菌体量を調べた。すると、30℃培養においてグルコースをまったく含まない培地では120時間の培養で10g湿重量/l弱が得られたが、グルコース濃度3%にすると約50g/lもの菌体を得ることができた。1%では35g/l程度、4%になると生育は阻害された。このように、YPD培地に含まれるグルコース濃度は、簡単な培地で多くのクロレラブルガリスNTM06を得るための重要な要素であることがわかった。(Example 4) Chlorella vulgaris NTM06 medium Chlorella vulgaris NTM06 in Example 1 grew best on a YPD medium. Therefore, the amount of cells obtained by changing the glucose concentration was examined. Then, in a medium containing no glucose at 30 ° C., a 10 g wet weight / l was obtained after 120 hours of cultivation, but about 50 g / l of cells could be obtained at a glucose concentration of 3%. At 1%, growth was inhibited at about 35 g / l and at 4%. Thus, it was found that the glucose concentration contained in the YPD medium is an important factor for obtaining many Chlorella vulgaris NTM06 with a simple medium.
以下に示す無機塩のみで、有機物を含まない培地(MBM培地)を調製し、片方はアルミ遮光し、もう一方は蛍光灯の光りを当てて30℃で7日間震盪培養したところ、光りを当てたほうは薄い緑色を呈し、遮光したほうは透明のままであった。このことは二酸化炭素固定能があることを示している。 Prepare a medium (MBM medium) that contains only the following inorganic salts and does not contain organic substances. One side is shielded from aluminum light, and the other is exposed to light from a fluorescent lamp and shaken at 30 ° C for 7 days. The one was light green and the one that was shielded from light remained transparent. This indicates that there is a carbon dioxide fixing ability.
MBM培地
x1 MBMmediumをオートクレーブ滅菌し、濾過滅菌したx100 Fe mixtureとx100 A5 metal mixtureをそれぞれ100分の1量加えて使用した。組成は以下に示す。
x1 MBM medium:
KNO325mg,MgSO4・7H2O7.5mg,K2HPO47.5mg,KH2PO417.5mg,NaCl2.5mg,CaCl2・2H2O1mg,Fe mixture0.1ml,A5 metal mixture0.1ml,
D.W.99.8ml,pH6.0
x100 Fe mixture:FeSO4・7H2O1g,D.W.500ml,H2SO4 two drops
x100 A5 metal mixture:H3BO3286mg,MnSO4・7H2O250mg,ZnSO4・7H2O 22.2mg,CuSO4・H2O7.9mg,Na2MoO42.1mg,D.W.100ml MBM medium x1 MBM medium was sterilized by autoclaving, and x100 Fe mixture and x100 A5 metal mix sterilized by filtration were added to each one-hundredth amount and used. The composition is shown below.
x1 MBM medium :
KNO 3 25 mg, MgSO 4 · 7H 2 O 7.5 mg, K 2 HPO 4 7.5 mg, KH 2 PO 4 17.5 mg, NaCl 2.5 mg, CaCl 2 · 2H 2 O 1 mg, Fe mixture 0.1 ml, A5 metal mix 0.1 ml ,
D. W. 99.8 ml, pH 6.0
x100 Fe mixture : FeSO 4 .7H 2 O 1 g, D.I. W. 500 ml, H 2 SO 4 two drops
x100 A5 metal mixture: H 3 BO 3 286mg, MnSO 4 · 7H 2 O250mg, ZnSO 4 · 7H 2 O 22.2mg, CuSO 4 · H 2 O7.9mg, Na 2 MoO 4 2.1mg, D. W. 100ml
単離されたクロレラブルガリスNTM06のMBM培地を用いた暗条件培養において、イーストナイトロジェンベースの添加(1%)では増殖は見られなかったが、グルコースを0.5%になるように添加すると増殖するようになった。暗条件における炭素源の資化性を以下に示す。以下の結果よりクロレラブルガリスNTM06はヘテロトロフィックな性質を有する。
グルコース、+
スクロース、+
マルトース、+
ガラクトース、+
可溶性でんぷん、+
カルボキシメチルセルロース、+
イヌリン、+
酢酸、+
セロビオース、−
ラクトース、−
キシロース、−
ラフィノース、−
ソルボース、−
メチルアルコール、−In dark culture using MBM medium of isolated Chlorella vulgaris NTM06, growth was not observed with the addition of yeast nitrogen base (1%), but when glucose was added to 0.5% It began to proliferate. The assimilation of the carbon source under dark conditions is shown below. From the following results, Chlorella vulgaris NTM06 has heterotrophic properties.
Glucose, +
Sucrose, +
Maltose, +
Galactose, +
Soluble starch, +
Carboxymethylcellulose, +
Inulin, +
Acetic acid, +
Cellobiose, −
Lactose,-
Xylose, −
Raffinose,-
Sorbose, −
Methyl alcohol, −
窒素源の利用性を調べるため、グルコースを1.0%になるように添加したMBM培地を調製した。クロレラブルガリスNTM06を培地に植え付け、暗条件培養にて、30℃、4日間、震盪培養し、増殖の有無を確かめた。以下に窒素源の利用性を示す。
KNO3+
NaNO3+
(NH4)Cl+
(NH4)2SO4+
Glycine+
なし +
上記のように、窒素源が存在しなくても弱い増殖が認められたので、クロレラブルガリスNTM06には窒素固定能が備わっていると考えられる。In order to examine the availability of the nitrogen source, an MBM medium supplemented with glucose at 1.0% was prepared. Chlorella vulgaris NTM06 was planted in the medium, and cultured under shaking in dark conditions at 30 ° C. for 4 days to confirm the presence or absence of proliferation. The availability of the nitrogen source is shown below.
KNO 3 +
NaNO 3 +
(NH 4 ) Cl +
(NH 4 ) 2 SO 4 +
Glycine +
None +
As described above, since weak growth was observed even in the absence of a nitrogen source, it is considered that Chlorella vulgaris NTM06 has a nitrogen fixing ability.
クロレラブルガリスNTM06はYPD平板培地を用いて酸素を除去した嫌気条件での生育を確認しているので、発酵または嫌気呼吸を行うこともできる。その場合は光照射が必要であるので、光合成と共役していると予想される。 Chlorella vulgaris NTM06 has been confirmed to grow under anaerobic conditions from which oxygen has been removed using a YPD flat plate medium. Therefore, fermentation or anaerobic respiration can also be performed. In that case, since light irradiation is required, it is expected to be conjugated with photosynthesis.
(実施例5)クロレラブルガリスNTM06を含む飼料添加物
アンモニア脱臭工程で得られたクロレラブルガリスNTM06は、簡単な水洗・水切りをして、あるいは、乾燥工程を経て得られたものを、飼料に添加することができる。例えば、培養されたクロレラブルガリスNTM06の懸濁液を遠心分離にかけ、水洗を繰り返した後、脱水する。脱水して得られた高濃度のクロレラブルガリスNTM06懸濁液を加熱処理して冷却した後、凍結乾燥させて乾燥クロレラブルガリスNTM06を得ることも可能である。なお、クロレラブルガリスNTM06の消化吸収を助けるため、細胞膜破砕加工を施すことも目的に応じて採用することができる。(Example 5) Feed additive containing Chlorella vulgaris NTM06 Chlorella vulgaris NTM06 obtained in the ammonia deodorization step is simply washed with water, drained, or obtained through a drying step. Can be added. For example, the cultured suspension of Chlorella vulgaris NTM06 is centrifuged, washed repeatedly with water, and then dehydrated. It is also possible to obtain a dried Chlorella vulgaris NTM06 by heat-treating and cooling the high-concentration Chlorella vulgaris NTM06 suspension obtained by dehydration and then freeze-drying it. In addition, in order to assist the digestion and absorption of Chlorella vulgaris NTM06, it is also possible to employ cell membrane disruption processing depending on the purpose.
本発明によれば、クロレラブルガリスNTM06を用いることで優れた脱臭能力を有するアンモニア脱臭法として有用である。また、大量の曝気やメタノールなどの炭素源の添加などの操作を必要としないので、操作や制御が容易でコストの低いアンモニア脱臭法として好適に用いられる。 According to the present invention, the use of Chlorella vulgaris NTM06 is useful as an ammonia deodorization method having excellent deodorization ability. Further, since it does not require an operation such as a large amount of aeration or addition of a carbon source such as methanol, it can be suitably used as an ammonia deodorization method that is easy to operate and control and low in cost.
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Cited By (3)
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CN105621795A (en) * | 2015-12-30 | 2016-06-01 | 中天正涵(天津)环保科技有限公司 | Method for removing odor substances from wastewater by utilization of deodorant biological bacteria |
JP2020114186A (en) * | 2019-01-18 | 2020-07-30 | 国立大学法人 宮崎大学 | Ammonia gas-resistant bacteria and ammonia gas-utilizing bacteria, and ammonia odor deodorization method using them |
WO2023188660A1 (en) * | 2022-03-29 | 2023-10-05 | 株式会社村田製作所 | Algal cultivation method, algal cultivation device, algal cultivation package, and algal slurry |
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
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CN105621795A (en) * | 2015-12-30 | 2016-06-01 | 中天正涵(天津)环保科技有限公司 | Method for removing odor substances from wastewater by utilization of deodorant biological bacteria |
JP2020114186A (en) * | 2019-01-18 | 2020-07-30 | 国立大学法人 宮崎大学 | Ammonia gas-resistant bacteria and ammonia gas-utilizing bacteria, and ammonia odor deodorization method using them |
JP7272551B2 (en) | 2019-01-18 | 2023-05-12 | 国立大学法人 宮崎大学 | Ammonia gas-tolerant bacteria, ammonia gas-utilizing bacteria, and ammonia odor deodorizing method using them |
WO2023188660A1 (en) * | 2022-03-29 | 2023-10-05 | 株式会社村田製作所 | Algal cultivation method, algal cultivation device, algal cultivation package, and algal slurry |
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