JPH0685712B2 - Hydrogen gas-producing bacteria - Google Patents
Hydrogen gas-producing bacteriaInfo
- Publication number
- JPH0685712B2 JPH0685712B2 JP29538390A JP29538390A JPH0685712B2 JP H0685712 B2 JPH0685712 B2 JP H0685712B2 JP 29538390 A JP29538390 A JP 29538390A JP 29538390 A JP29538390 A JP 29538390A JP H0685712 B2 JPH0685712 B2 JP H0685712B2
- Authority
- JP
- Japan
- Prior art keywords
- strain
- hydrogen gas
- clostridium
- strains
- producing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 title claims description 66
- 241000894006 Bacteria Species 0.000 title description 25
- 241000256602 Isoptera Species 0.000 claims description 17
- 238000011160 research Methods 0.000 claims description 11
- 241000193403 Clostridium Species 0.000 claims description 8
- 230000001580 bacterial effect Effects 0.000 claims description 3
- 241000894007 species Species 0.000 claims 1
- 244000005700 microbiome Species 0.000 description 29
- 238000004519 manufacturing process Methods 0.000 description 23
- 239000007789 gas Substances 0.000 description 19
- 235000000346 sugar Nutrition 0.000 description 11
- 230000000694 effects Effects 0.000 description 8
- 239000002609 medium Substances 0.000 description 8
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 8
- 238000000034 method Methods 0.000 description 8
- 239000002699 waste material Substances 0.000 description 8
- 238000009776 industrial production Methods 0.000 description 7
- 230000000243 photosynthetic effect Effects 0.000 description 7
- 239000002351 wastewater Substances 0.000 description 7
- 229920001817 Agar Polymers 0.000 description 5
- 239000008272 agar Substances 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 239000001257 hydrogen Substances 0.000 description 5
- 229910052739 hydrogen Inorganic materials 0.000 description 5
- 241000588915 Klebsiella aerogenes Species 0.000 description 4
- 229940092559 enterobacter aerogenes Drugs 0.000 description 4
- 239000002803 fossil fuel Substances 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 150000008163 sugars Chemical class 0.000 description 4
- 241001148471 unidentified anaerobic bacterium Species 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000002485 combustion reaction Methods 0.000 description 3
- 238000003912 environmental pollution Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000004065 wastewater treatment Methods 0.000 description 3
- 239000002028 Biomass Substances 0.000 description 2
- 241000193171 Clostridium butyricum Species 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- ATUOYWHBWRKTHZ-UHFFFAOYSA-N Propane Chemical compound CCC ATUOYWHBWRKTHZ-UHFFFAOYSA-N 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 230000000593 degrading effect Effects 0.000 description 2
- 239000010840 domestic wastewater Substances 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 238000009434 installation Methods 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000010865 sewage Substances 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- GYYDPBCUIJTIBM-DYOGSRDZSA-N (2r,3s,4s,5r)-2-(hydroxymethyl)-6-[[(4r,5s)-4-hydroxy-3-methyl-2,6-dioxabicyclo[3.2.1]octan-8-yl]oxy]-4-methoxyoxane-3,5-diol Chemical compound O[C@@H]1[C@@H](OC)[C@@H](O)[C@@H](CO)OC1OC1[C@H]2OCC1OC(C)[C@H]2O GYYDPBCUIJTIBM-DYOGSRDZSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 206010003497 Asphyxia Diseases 0.000 description 1
- 241000589151 Azotobacter Species 0.000 description 1
- 235000016068 Berberis vulgaris Nutrition 0.000 description 1
- 241000335053 Beta vulgaris Species 0.000 description 1
- 241000193454 Clostridium beijerinckii Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000588722 Escherichia Species 0.000 description 1
- BDAGIHXWWSANSR-UHFFFAOYSA-M Formate Chemical compound [O-]C=O BDAGIHXWWSANSR-UHFFFAOYSA-M 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102100024295 Maltase-glucoamylase Human genes 0.000 description 1
- 241000192520 Oscillatoria sp. Species 0.000 description 1
- 206010035664 Pneumonia Diseases 0.000 description 1
- 241000191021 Rhodobacter sp. Species 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 239000004280 Sodium formate Substances 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 241000204046 Termitidae Species 0.000 description 1
- 238000003916 acid precipitation Methods 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 125000003158 alcohol group Chemical group 0.000 description 1
- 102000005840 alpha-Galactosidase Human genes 0.000 description 1
- 108010030291 alpha-Galactosidase Proteins 0.000 description 1
- 108010028144 alpha-Glucosidases Proteins 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000000721 bacterilogical effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 239000003034 coal gas Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000005868 electrolysis reaction Methods 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 239000003345 natural gas Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000001294 propane Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- HLBBKKJFGFRGMU-UHFFFAOYSA-M sodium formate Chemical compound [Na+].[O-]C=O HLBBKKJFGFRGMU-UHFFFAOYSA-M 0.000 description 1
- 235000019254 sodium formate Nutrition 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000005979 thermal decomposition reaction Methods 0.000 description 1
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 description 1
- 229920002554 vinyl polymer Polymers 0.000 description 1
- 238000010792 warming Methods 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
【発明の詳細な説明】 〔産業上の利用の分野〕 本発明は、微生物、特に水素ガスの産生菌に関するもの
であり、詳しくは、シロアリより単離された新規な水素
ガス産生菌を提供するものである。TECHNICAL FIELD The present invention relates to a microorganism, particularly a hydrogen gas-producing bacterium, and in particular, provides a novel hydrogen gas-producing bacterium isolated from termites. It is a thing.
本発明に係る微生物は、水素ガス産生能にすぐれている
ので、水素ガスの工業生産に有用であり、したがって本
発明は、エネルギーの技術分野において大きな貢献をな
すものであるが、そのうえ更に、本発明に係る微生物
は、各種の糖を広範に且つ強力に分解する能力も併有し
ているので、食品工業や製紙工業由来の廃水や生活廃水
等特に糖類を多量に含有する廃水の処理に有用であり、
廃水処理ないし公害防止の技術分野においても多大の貢
献をなすものである。Since the microorganism according to the present invention is excellent in hydrogen gas producing ability, it is useful for industrial production of hydrogen gas, and thus the present invention makes a great contribution in the technical field of energy, and furthermore, Since the microorganism according to the invention has a capability of decomposing various sugars extensively and strongly, it is useful for treating wastewater containing a large amount of sugars, such as wastewater and domestic wastewater derived from the food industry and paper industry. And
It also makes a great contribution to the technical field of wastewater treatment or pollution control.
現代工業社会においては、石油、石炭、天然ガスなどの
化石燃料が大量に消費され、その化石燃料は、燃焼によ
り多量のNOx、SOxおよびCO2などを排出し、その結果、
環境汚染、酸性雨、地球の温暖化などの諸問題を惹起し
ている。更には、その埋蔵量が有限で近い将来枯渇する
といわれ、重要な社会問題ともなっている。In the modern industrial society, fossil fuels such as oil, coal, and natural gas are consumed in large amounts, and the fossil fuels emit large amounts of NOx, SOx, CO 2 and the like by combustion, and as a result,
It causes various problems such as environmental pollution, acid rain, and global warming. Furthermore, it is said that the reserves are limited and will be exhausted in the near future, which is an important social problem.
これらのことから、化石燃料に代わる環境汚染のない新
しいクリーンなエネルギー源が世界的に求められてお
り、石油に代わる次世代のエネルギー源として、現在、
アルコール及びメタンガスが注目されている。しかし、
アルコールやメタンガスは、いずれも燃焼により大量に
CO2を産生する点では、依然として問題があり、しか
も、その内在エネルギーはロケットや航空機用の燃料に
使用し得るほど高いものではないという欠点を有してい
る。For these reasons, there is a global demand for a new clean energy source that replaces fossil fuels without environmental pollution, and is currently being used as a next-generation energy source that replaces petroleum.
Alcohol and methane gas are receiving attention. But,
Alcohol and methane gas are burned in large quantities
In terms of producing CO 2, there is still a problem, moreover, its inherent energy has the disadvantage that not higher may be used as fuel for rocket and aircraft.
そこで、水素ガスが注目されるようになった。水素ガス
は、単位重量当りの燃焼による発熱エネルギーが石油の
３倍もあり、しかも、燃焼による副生物がH2Oのみであ
ることから、次世代の理想的なクリーンエネルギー源と
して期待されるからである。Then, hydrogen gas came to the attention. Hydrogen gas, exothermic energy by combustion per unit weight is also three times the oil, moreover, since the by-products due to combustion is only H 2 O, since it is expected as an ideal clean energy source in the next generation Is.
しかしながら、現状での水素ガスの工業的製法は、水の
電気分解や液化プロパン（LPG）、アルコールの高圧熱
分解などの方法によっているため、これらの方法は、そ
のエネルギー源として化石燃料を消費するものであるか
ら、製造法におけるエネルギー源の問題が解決されない
限り前述した環境汚染などの諸問題の基本的解決にはな
らない。However, the current industrial manufacturing method of hydrogen gas depends on the methods such as electrolysis of water, liquefied propane (LPG), and high-pressure thermal decomposition of alcohol. Therefore, these methods consume fossil fuel as the energy source. Therefore, unless the problem of the energy source in the manufacturing method is solved, it does not become a fundamental solution to the above-mentioned problems such as environmental pollution.
そこで微生物が着目され、微生物による水素ガスの産生
に関する研究がいくつか試みられてきた。たしかに、微
生物により水素ガスを生産するという方法が確立される
とすれば、その方法の利点は、反応が常温、常圧で行な
われるから、システム構成が簡単であり、また、エネル
ギー消費も極めて少ないということであり、しかも、再
生可能なバイオマスを水素ガス産生の原料として使用す
るものであって、このバイオマスはもともと太陽エネル
ギーを変換したものであるので自然エネルギーの有効利
用であることになる。更にまた、微生物による水素ガス
の産生には、通常、廃棄物または廃液中に存在する有機
物質を原料とすることが可能であるので、廃液の効率的
処理による環境浄化の問題の解決にもなるという利点が
ある。Therefore, attention has been paid to microorganisms, and several studies on the production of hydrogen gas by microorganisms have been attempted. Certainly, if a method of producing hydrogen gas by microorganisms is established, the advantage of this method is that the reaction is carried out at room temperature and atmospheric pressure, so the system configuration is simple and energy consumption is extremely low. This means that renewable biomass is used as a raw material for hydrogen gas production. Since this biomass is originally converted from solar energy, it is an effective use of natural energy. Furthermore, for the production of hydrogen gas by microorganisms, it is usually possible to use organic substances present in wastes or waste liquids as raw materials, and therefore it is possible to solve the problem of environmental purification by efficient treatment of waste liquids. There is an advantage.
上記のように微生物による水素ガスの産生に関する研究
がいくつか行われた結果、水素ガスを産生する微生物が
若干発見された。As a result of several studies on the production of hydrogen gas by microorganisms as described above, some microorganisms that produce hydrogen gas have been discovered.
これらのこれまでに知られている、水素ガスを産生する
微生物は、大別すると、光合成微生物と非光合成細菌と
に分けられる。前者には、光合成細菌のRhodobacter sp
haeroides、藍藻のOscillatoria sp.Miami BG7があり、
後者には、窒素固定細菌のAzotobacter chroococuum,Kl
ebsiella pneumonia、通性嫌気性細菌のEscherichia co
li,Enterobacter aerogenes、嫌気性細菌のClostridium
butyricum等がある。These known hydrogen gas-producing microorganisms are roughly classified into photosynthetic microorganisms and non-photosynthetic bacteria. The former includes the photosynthetic bacterium Rhodobacter sp.
haeroides, blue-green alga Oscillatoria sp. Miami BG7,
The latter includes the nitrogen-fixing bacteria Azotobacter chroococuum, Kl.
ebsiella pneumonia, a facultative anaerobic bacterium Escherichia co
li, Enterobacter aerogenes, an anaerobic bacterium Clostridium
butyricum etc.
たしかに、このように微生物による水素ガスの産生は、
水素ガスの製造方法として、極めて優れた利点を有して
はいるが、従来の研究業績においては、未だそれを工業
的な製造方法として確立するにはほど遠い状況にある。
特に、これまでに行われている研究では、工業的な生産
を可能にするほどの水素ガスの生産性の高い微生物は見
出されておらず、したがって、現状では微生物を利用し
て水素ガスを工業的に製造する方法については、全く未
開発の状況にある（福井三郎ほか監修「バイオテクノロ
ジー事典」（株）シーエムシー（1986-10-9）p.601-60
2）。Certainly, the production of hydrogen gas by microorganisms is
Although it has an extremely excellent advantage as a method for producing hydrogen gas, it is still far from being established as an industrial production method in the past research achievements.
In particular, research conducted to date has not found a microorganism with high hydrogen gas productivity that enables industrial production, and therefore, at present, the microorganism is used to generate hydrogen gas. The industrial production method is completely undeveloped (Saburo Fukui et al. "Biotechnology Encyclopedia" CMC Co., Ltd. (1986-10-9) p.601-60
2).
上記のように微生物による水素ガスの産生技術は、未だ
工業的レベルにまでは達していない。As described above, the technology for producing hydrogen gas by microorganisms has not yet reached the industrial level.
従来既知の微生物は、いずれも、水素ガスの産生効率自
体が低いだけでなく、これらの微生物の内、光合成微生
物については、それによる水素ガスの産生には、光エネ
ルギーを利用するために、表面積の広い培養槽と多量の
水を必要とする。All of the conventionally known microorganisms are not only low in hydrogen gas production efficiency, but among these microorganisms, photosynthetic microorganisms are required to use light energy to produce hydrogen gas. Requires a large culture tank and a large amount of water.
他方、非光合成細菌による水素ガスの産生は、小規模の
発酵槽によっても可能であり、地下に設置するなど、そ
の設置場所の選択肢が広いなどの利点があり、水素ガス
の産生には、非光合成細菌による方が光合成微生物によ
るよりも有利であると考えられる。On the other hand, the production of hydrogen gas by non-photosynthetic bacteria is possible even with a small-scale fermenter, and there are advantages such as a wide choice of installation locations such as installation in the underground. It is considered that photosynthetic bacteria are more advantageous than photosynthetic microorganisms.
現在までに単離された微生物のなかで、最も効率よく水
素ガスを産生する微生物は、Tanisho S.らが単離したエ
ンテロバクター アロエゲネス（Enterobacter aerogen
es）E82005株であるとされている（Tanisho S.,et al.l
nt.J.Hydrogen Energy 12 623,1987;Biochim.Biophys.A
cta.973 1 1989）。しかし、この菌株は、通性嫌気性細
菌であり、嫌気状態でも増殖するが、好気性条件下でよ
り活発に増殖するため、発酵槽内で大量の水素が産生さ
れると、槽内を好気的に維持することが困難になり、水
素ガスの工業的生産には適さない。Among the microorganisms isolated to date, the one that produces hydrogen gas most efficiently is Enterobacter aerogenes (Enterobacter aerogenes) isolated by Tanisho S.
es) E82005 strain (Tanisho S., et al.l)
nt.J.Hydrogen Energy 12 623,1987; Biochim.Biophys.A
cta.973 1 1989). However, this strain is a facultative anaerobic bacterium and grows even in an anaerobic state, but since it grows more actively under aerobic conditions, when a large amount of hydrogen is produced in the fermenter, the strain in the fermentor is favored. It becomes difficult to maintain in a gas state and is not suitable for industrial production of hydrogen gas.
本発明の目的は、工業的生産に好適な水素ガス産生菌を
新たに開発するとともに、廃水、特に糖類に富む廃水を
効率よく処理しうる微生物を新たに開発することであ
る。An object of the present invention is to newly develop a hydrogen gas-producing bacterium suitable for industrial production, and also to newly develop a microorganism capable of efficiently treating wastewater, particularly saccharide-rich wastewater.
本発明は、上記目的を達成するためになされたものであ
って、各方面から鋭意検討したにもかかわらず成功する
には至らず、目的を達成しうる微生物をスクリーニング
するには、従来からの発想を大転換する必要があること
を認めた。The present invention has been made in order to achieve the above-mentioned object, and it has not been successful in spite of diligent studies from various directions. I admitted that it is necessary to change the way of thinking.
そこで、微生物の起源について本発明者らは、鋭意研究
を重ねた結果、水素ガス産生細菌の起源を過去試みられ
たことのない昆虫に求め、遂にシロアリ（Termites for
mosans）より、極めて反応速度が速く、かつ大量の水素
ガスを産生する新規な細菌を単離することに成功した。
そして更に驚くべきことに、これらの微生物は糖を分解
する能力が非常に高いことも併せ確認し、これらの有効
な新知見を基礎として本発明が完成されたのである。Therefore, as a result of intensive studies on the origin of microorganisms, the present inventors have sought the origin of hydrogen gas-producing bacteria from an insect that has never been tried, and finally found termites (Termites for
mosans) has succeeded in isolating a new bacterium that has a very high reaction rate and produces a large amount of hydrogen gas.
And, surprisingly, it was also confirmed that these microorganisms have a very high ability to decompose sugar, and the present invention was completed based on these effective new findings.
すなわち本発明は、シロアリ、つまり等翅目、シロアリ
科（Termitidae）に属する昆虫由来の水素ガス産生菌に
関する発明を基本的技術思想とするものである。That is, the present invention has as its basic technical idea an invention relating to a termite, that is, an insect-derived hydrogen gas-producing bacterium belonging to the order Heptoptera, Termitidae.
更に詳細には、本発明は、シロアリ由来の、水素ガス産
生能がきわめて高い水素ガス産生菌に関するものであっ
て、例えば下記するような分離菌を包含する。（これら
の水素ガス産生菌は、いずれも工業技術院微生物工業技
術研究所（現、生命工学工業技術研究所）に寄託されて
いる。） AM14A-2株（微工研菌寄第11793号）、AM37E株（同、第1
1794号）、AM21B株（微工研条寄第3592号）、AM9A-2株
（微工研菌寄第第11800号）、AM37F株（微工研条寄第35
93号）、AM40A株（微工研菌寄第第11798号）、AM18B株
（同、第11796号）、AM38C-1株（同、第11797号）およ
びAM42E株（同、第11801号）。More specifically, the present invention relates to a hydrogen gas-producing bacterium, which is derived from termites and has an extremely high hydrogen gas-producing ability, and includes, for example, the following isolates. (All of these hydrogen gas-producing bacteria have been deposited at the Institute of Microbial Science and Technology of the Agency of Industrial Science and Technology (currently the Institute of Biotechnology and Industrial Technology).) AM14A-2 strain (Microtechnology Research Institute No. 11793) , AM37E strain (the same, first
1794), AM21B strain (MICR 3592), AM9A-2 strain (MICR 11800), AM37F (MICR 35)
93), AM40A strain (Microtechnology Research Institute, No. 11798), AM18B strain (No. 11796), AM38C-1 strain (No. 11797) and AM42E strain (No. 11801).
以下、本発明を詳しく説明する。Hereinafter, the present invention will be described in detail.
本発明を実施するには、まずシロアリから嫌気性菌を分
離する必要がある。それには、例えばイエシロアリ（Te
rmites formosans）を生きた状態のままで窒息死させ、
酢酸塩及びギ酸塩（塩としてはアルカリ（土）金属塩が
好適）を含有する分離用培地を用いて目的とする微生物
を分離する。In order to carry out the present invention, it is first necessary to isolate anaerobic bacteria from termites. For example, the termite (Te
rmites formosans), suffocate and kill
A target microorganism is separated using a separation medium containing an acetate and a formate (alkaline (earth) metal salts are preferable as the salt).
例えば、本発明者らは、細菌の分離培養用培地として、
普通ブイヨン（日水製薬株式会社製）を通常の50分の１
に希釈し培養液（以下1/50Nと略記する）にメタンガス
の基質である酢酸ソーダと水素ガスの基質である蟻酸ソ
ーダとをそれぞれ2.5g/づつ加えて調製した培養液
（以下1/50N+と略記する）を考案して用いた。また、こ
の1/50N+に1.5％の割に寒天を加えた貧栄養1/50N+寒天
培地を創案した。次に培養条件として、将来エネルギー
回収型の廃液処理にも応用することを考慮し、35℃での
嫌気性培養を用いることにした。For example, the inventors of the present invention, as a culture medium for bacterial isolation culture,
Ordinary broth (manufactured by Nissui Pharmaceutical Co., Ltd.)
The diluted culture (more than 1 / abbreviated to 50 N) in the sodium formate respectively 2.5 g / increments added was prepared broth is a substrate of sodium acetate and hydrogen gas which is a substrate for methane gas (less than 1/50 N + Abbreviated as). In addition, we created an oligotrophic 1 / 50N + agar medium in which agar was added to 1.5% of this 1 / 50N + . Next, we decided to use anaerobic culture at 35 ℃ as a culture condition, considering that it will be applied to waste liquid treatment of energy recovery type in the future.
イエシロアリから嫌気性細菌を効率よく分離培養するた
めに、極力酸素との接触を避けることに留意し、そのた
めに、巣より取り出した生きているイエシロアリをペト
リー皿に入れ、それを嫌気性培養器（グローブボック
ス、米国フォーマ社製）内に納め、嫌気性混合ガス（H2
10％、CO210％、N280％）の雰囲気下で窒息死させた。
このイエシロアリをすりつぶすことなく、シロアリ10匹
を1/50N+寒天培地20mlに加え、充分に混和して固化させ
た。この操作によって、シロアリが寒天中に埋設した状
態となり混合ガスにもあまり接触しない条件が作り出さ
れた。In order to efficiently separate and culture the anaerobic bacteria from the termites, keep in mind that contact with oxygen should be avoided as much as possible, and for that purpose, living termites taken out from the nest should be placed in a Petri dish and placed in an anaerobic incubator ( It is stored in a glove box, made by U.S. former company, and anaerobic mixed gas (H 2
They were killed by suffocation in an atmosphere of 10%, CO 2 10%, N 2 80%).
Without crushing the termites, 10 termites were added to 1 / 50N + 20 ml of agar medium and mixed sufficiently to solidify. This operation created a condition in which the termites were buried in agar and did not contact the mixed gas too much.
この寒天平板10枚を嫌気性混合ガスの雰囲気下で35℃で
３週間培養を行った。Ten of these agar plates were cultured at 35 ° C. for 3 weeks in an anaerobic mixed gas atmosphere.
次いで、細菌学的な分離操作により153株の細菌を単離
し、その153株の細菌の水素ガス産生能をスクリーニン
グ法により検定した結果、93％に相当する。141株が水
素ガスを産生することが見出された。次いで酸素要求性
試験を行った結果、８株が通性嫌気性細菌であった。最
終的には、目的に適う候補菌37株の分離に成功した。Next, 153 strains of bacteria were isolated by a bacteriological separation operation, and the hydrogen gas producing ability of the 153 strains was assayed by a screening method, which corresponds to 93%. 141 strains were found to produce hydrogen gas. Then, as a result of an oxygen requirement test, 8 strains were facultative anaerobic bacteria. Finally, we succeeded in isolating 37 strains of candidate strains suitable for the purpose.
現在までに分離された微生物のなかで、最も効率よく水
素ガスを産生する微生物は、前述したとおり、Enteroba
cter aerogenes E82005株とされているが、この菌株は1
1mmol H2/‐medium.hr又は246ml H2/‐medium.hrの
水素ガスを産生するのに対し、後記するところからも明
らかなように本発明者が単離した菌株のうち、９株は、
これをはるかに超える水素ガス産生能を有する点できわ
めて特徴的である。Among the microorganisms that have been isolated to date, the one that produces hydrogen gas most efficiently is Enteroba as described above.
cter aerogenes E82005 strain, but this strain is 1
In contrast to producing hydrogen gas of 1 mmol H 2 /-medium.hr or 246 ml H 2 /-medium.hr, 9 strains among the strains isolated by the present inventor are clear as will be apparent from the later description. ,
It is extremely characteristic in having a hydrogen gas production capacity far exceeding this.
これら９株の一般的性状、生化学的性状、酵素活性を表
１−１、表１−２及び表１−３に示す。The general properties, biochemical properties and enzyme activity of these 9 strains are shown in Table 1-1, Table 1-2 and Table 1-3.
これらの各菌株の性状（API 20A,Rap ANA II systemに
よる）より、AM21BはClostridium beijerinckii,AM37E
とAM14A-2はClostridium butyricumと同定された。AM38
C-1,AM42E及びAM18Bの３菌株はClostridium属に属する
と認められるが、このように高い水素ガス産生能を有す
る微生物は知られておらず、またシロアリ由来である点
も考慮に入れて、これらを新規な細菌と同定し、それぞ
れ、Clostridium AM38C-1、Clostridium AM42E、及びCl
ostridium 18Bと命名した。 From the properties of each of these strains (API 20A, Rap ANA II system), AM21B was Clostridium beijerinckii, AM37E.
And AM14A-2 were identified as Clostridium butyricum. AM38
Three strains of C-1, AM42E and AM18B are recognized as belonging to the genus Clostridium, but no microorganisms having such high hydrogen gas-producing ability are known, and taking into consideration that they are derived from termites, We identified these as novel bacteria and identified them with Clostridium AM38C-1, Clostridium AM42E, and Cl, respectively.
It was named ostridium 18B.
さらに、AM37F、AM9A-2及びAM40Aの３株は、同定不能な
全く新しい水素産生菌である。更に詳細に言えば、AM37
F、AM9A-2及びAM40Aのように、グラム陽性、桿菌、無芽
胞性嫌気性細菌で水素ガスを産生する細菌については、
これまで全く報告されておらず、例をみないものであ
る。これらの３菌株は、世界で初めて見出された新規細
菌である。Furthermore, the three strains AM37F, AM9A-2 and AM40A are completely new hydrogen-producing bacteria that cannot be identified. In more detail, AM37
For F, AM9A-2 and AM40A, for Gram-positive, bacilli, and asporogenic anaerobic bacteria that produce hydrogen gas,
It has never been reported so far, and it is an example. These three strains are new bacteria found for the first time in the world.
そこで、既述した菌学的諸性質を検討し、特に従来のCl
ostridium属とは、シロアリを由来とする点、水素産生
能がきわめて高い点、糖分解能が高い点、芽胞形成能を
欠く点等で決定的且つ明確に区別できることから、これ
らを新規な属に属せしめるのが相当と認め、新たにテル
ミテバクター（Termitebacter）属を創設し、これらの
３菌株をこの新属に属せしめ、Termitebacter AM37F、T
ermitebacter AM9A-2、Termitebacter AM40Aとそれぞれ
命名した。Therefore, we examined the various mycological properties described above, and especially the conventional Cl
The genus ostridium is a genus that belongs to a new genus because it can be decisively and clearly distinguished from the termite origin, the extremely high hydrogen-producing ability, the high sugar decomposing ability, and the lack of spore-forming ability. Recognizing that it is appropriate to allow them, a new genus Termitebacter was created, and these three strains were made to belong to this new genus, Termitebacter AM37F, T
ermitebacter AM9A-2 and Termitebacter AM40A were named respectively.
これら菌株のガム寒天の高層斜面培地におけるガス産生
性能を表Ｉ−４に示す。The gas production performance of gum agar of these strains in the high-rise slant medium is shown in Table I-4.
また、人工汚水におけるガス産生能を表１−５に示す。 Table 1-5 shows the gas production capacity of artificial sewage.
ガス産生量の測定は、添付図面第１図に示す装置を用い
て行った。 The amount of gas produced was measured using the apparatus shown in FIG.
すなわち、培養ビンの口を密栓したゴム栓に18号注射針
を刺し通し、そこにビニールチューブをつなぎ、この排
気管をガス測定用シリンダーに接続した。培養後、逆さ
にしたシリンダーに溜まったガス産生量を測定した。That is, a No. 18 injection needle was pierced through a rubber stopper in which the mouth of a culture bottle was sealed, a vinyl tube was connected there, and this exhaust pipe was connected to a gas measuring cylinder. After the culture, the amount of gas produced in the inverted cylinder was measured.
前記したとおり、Enterobacter aerogenes E82005株の
水素ガス産生能は246ml/・hrであるのに対し、本発明
者がイエシロアリより分離に成功した前記の９菌株の水
素産生能は、表１−６に示すようにE82005株と比較して
全く比較にならないほど大量の水素ガスを産生し、従来
菌に比してその10倍以上も産生する菌株も認められ、こ
れらの菌株は工業的用途に充分使用可能であることが実
証された。As described above, the hydrogen gas producing ability of the Enterobacter aerogenes E82005 strain is 246 ml / .hr, whereas the hydrogen producing ability of the 9 strains successfully separated by the present inventor from the termite is shown in Table 1-6. As can be seen, there are strains that produce a large amount of hydrogen gas that is incomparable to the E82005 strain, and produce more than 10 times that of conventional strains, and these strains can be used for industrial purposes sufficiently. Was proved.
次に、上記の９菌株の糖分解能と糖分解系酵素活性をみ
ると、非常に多種多様な活性を有していることが判明し
た（表１−２、表１−３参照）。 Next, the sugar degrading and sugar degrading enzyme activities of the above 9 strains were found to have a very wide variety of activities (see Table 1-2 and Table 1-3).
特に、上記の９菌株は、いずれも、α‐glucosidase活
性及びα‐galactosidase活性が強力であり、とりわ
け、AM9A-2、AM37F、AM14A-2及びAM37Eの４株の酵素活
性は卓越していることが判明した。In particular, all of the above 9 strains have strong α-glucosidase activity and α-galactosidase activity, and in particular, AM9A-2, AM37F, AM14A-2 and AM37E 4 strains have excellent enzyme activity. There was found.
これらの結果からして、糖を含有するないし糖を成分と
する天然物を処理するに当って、これらの菌株は非常に
有効であって、ジュース中各種農産物の清澄化、低粘
化、流動化に利用することができるのみでなく、有機廃
水、例えばビート工場やオレンジ加工工場その他パルプ
植物繊維、殿粉あるいは多糖類などを含む廃液を処理す
るにあたって、これらの菌株は特に有効な細菌株である
ということができ、廃液・廃棄物の処理による環境浄化
に利用することができ、そして更に、水素ガスの製造の
工業化においてこれらの菌株は極めて重要な役割を果た
すものである。Based on these results, these strains are very effective in processing sugar-containing or sugar-containing natural products, and are effective in clarifying, reducing viscosity, and reducing flow of various agricultural products in juice. These strains are particularly effective bacterial strains in treating organic wastewater, such as beet mills, orange processing mills, and other pulp plant fibers, starch or polysaccharides, etc. It can be said that it can be used for environmental purification by treating waste liquid / waste, and furthermore, these strains play an extremely important role in the industrialization of hydrogen gas production.
本発明者らの見出した新規な水素ガス産生菌は、極めて
速い反応速度で、かつ時間あたり極めて大量の水素ガス
を産生するという画期的な特徴的性格を有するものであ
り、それとともに食品工業・製紙工業などに由来する廃
水、あるいは生活廃水あるいはあらゆる有機物質廃棄物
などの処理にとっても極めて有効な細菌である。また、
各種廃水を処理すると同時に水素ガスを産生させるこ
と、つまり廃水処理と水素ガスの産生とを同時に行うの
にも、本発明に係る微生物は極めて有効に利用すること
ができるのである。The novel hydrogen gas-producing bacterium found by the present inventors has an epoch-making characteristic character of producing an extremely large amount of hydrogen gas per hour at an extremely fast reaction rate, and together with it, the food industry.・ It is an extremely effective bacterium for the treatment of wastewater derived from the paper industry, domestic wastewater, and all organic substance wastes. Also,
The microorganisms according to the present invention can be extremely effectively used for treating various kinds of wastewater and producing hydrogen gas at the same time, that is, for simultaneously performing the wastewater treatment and the production of hydrogen gas.
次に本発明の実施例を示す。Next, examples of the present invention will be described.
実施例１ グルコースをそれぞれ、0.3％及び1.0％加えた普通ブイ
ヨン（日水製薬製）にAM21B菌（FERM BP-3592）を接種
し、第１図に示す装置を用いて、36℃の水槽で培養し、
発生する水素ガスの量を１時間毎に測定した。結果は表
１−７に示すとおりであった。Example 1 AM21B bacterium (FERM BP-3592) was inoculated into ordinary broth (manufactured by Nissui Pharmaceutical Co., Ltd.) to which glucose was added at 0.3% and 1.0%, respectively, and the same apparatus as shown in FIG. Cultivate,
The amount of hydrogen gas generated was measured every hour. The results are shown in Table 1-7.
実施例２ 人工汚水からの水素ガスの産生 0.3％グルコース加普通ブイヨン（日水製薬製）とガム
ブイヨン培地（日水製薬製）の各300mlに各菌を接種し
て、36℃で１夜静置培養した。産生されたガス量と水素
ガス濃度より水素ガス産生量を算出した。結果は表１−
８に示すとおりであった。 Example 2 Production of hydrogen gas from artificial sewage 300 ml of 0.3% glucose-added normal broth (manufactured by Nissui Pharmaceutical) and 300 ml of gum broth medium (manufactured by Nissui Pharmaceutical) were inoculated with each bacterium and left overnight at 36 ° C. The culture was performed. The hydrogen gas production amount was calculated from the produced gas amount and the hydrogen gas concentration. The results are shown in Table 1-
It was as shown in 8.
実施例３ 新規な水素ガス産生菌による水素ガス産生 人工汚染としてのガムブイヨン（日水製薬製）培地900m
lに各培養菌液100mlを添加し撹拌して培養した。１時間
毎にガス産生量、菌量（OD）、pH及び糖濃度を測定し、
ガス産生量が200ml以上の場合のガスの組成分析を行っ
た。ガス産生量とガス濃度から、水素ガスの産生量を算
出し、培養時間と水素ガス産生量の関係を検討した。表
１−９はその結果を示すものである。 Example 3 Production of hydrogen gas by a novel hydrogen gas-producing bacterium Gum broth (manufactured by Nissui Pharmaceutical) medium 900 m as artificial contamination
100 ml of each culture broth was added to 1 and stirred to culture. Measure the amount of gas produced, the amount of bacteria (OD), pH and sugar concentration every hour,
The composition of the gas was analyzed when the gas production was 200 ml or more. The production amount of hydrogen gas was calculated from the gas production amount and the gas concentration, and the relationship between the culture time and the hydrogen gas production amount was examined. Table 1-9 shows the result.
〔発明の効果〕 以上述べたところから明らかなように、本発明に係るシ
ロアリ由来の新規な水素ガス産生菌は、きわめて水素ガ
ス産生能が高いのみでなく、有機物とりわけ単糖、少
糖、多糖類等各種の糖類の分解能にもすぐれているとい
う特徴を有するものである。 EFFECTS OF THE INVENTION As is clear from the above description, the novel termite-derived hydrogen gas-producing bacterium according to the present invention has not only a very high hydrogen gas-producing ability, but also organic substances, particularly monosaccharides, oligosaccharides, and polysaccharides. It has a feature that it is also excellent in decomposing various sugars such as sugars.
したがって、本発明に係る新規な水素ガス産生菌は、水
素ガスの工業的製造法及び／又は廃水処理、廃棄物処理
に有用な優れた活性を有し、産業上極めて価値ある微生
物である。Therefore, the novel hydrogen gas-producing bacterium according to the present invention is an extremely valuable industrial microorganism having an excellent activity useful for industrial production method of hydrogen gas and / or wastewater treatment and waste treatment.
添付第１図は、本発明に係る新規微生物のガス産生量を
測定するために使用した装置の１例を示すものである。FIG. 1 attached herewith shows an example of an apparatus used for measuring the gas production of a novel microorganism according to the present invention.
───────────────────────────────────────────────────── フロントページの続き (72)発明者 鷹野 幹雄 東京都調布市飛田給２丁目19番１号 鹿島 建設株式会社技術研究所内 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Mikio Takano 2-19-1 Tobita-Su, Chofu-shi, Tokyo Kashima Construction Co., Ltd. Technical Research Institute
Claims (2)
を有する次の菌種： クロストリジウム・ベイエリンキイ（Clostridium beij
erinckii）、 クロストリジウム・AM18B、 クロストリジウム・AM38C-1、 クロストリジウム・AM42E テルミテバクター・AM9A-2、 テルミテバクター・AM37Fまたは テルミテバクター・AM40A。1. The following bacterial species derived from termites and having a high hydrogen gas-producing ability: Clostridium beijinkyi (Clostridium beij)
erinckii), Clostridium AM18B, Clostridium AM38C-1, Clostridium AM42E Thermitebactor AM9A-2, Thermitebactor AM37F or Thermitebactor AM40A.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP29538390A JPH0685712B2 (en) | 1990-11-02 | 1990-11-02 | Hydrogen gas-producing bacteria |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP29538390A JPH0685712B2 (en) | 1990-11-02 | 1990-11-02 | Hydrogen gas-producing bacteria |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH04169178A JPH04169178A (en) | 1992-06-17 |
| JPH0685712B2 true JPH0685712B2 (en) | 1994-11-02 |
Family
ID=17819917
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP29538390A Expired - Lifetime JPH0685712B2 (en) | 1990-11-02 | 1990-11-02 | Hydrogen gas-producing bacteria |
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| Country | Link |
|---|---|
| JP (1) | JPH0685712B2 (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6978572B1 (en) | 1998-11-06 | 2005-12-27 | Colorado State University Research Foundation | Method and device for attracting insects |
| JP3898454B2 (en) | 2001-03-06 | 2007-03-28 | シャープ株式会社 | Polymer electrolyte fuel cell |
| US7432091B2 (en) | 2003-02-24 | 2008-10-07 | Research Institute Of Innovative Technology For The Earth | Highly efficient hydrogen production method using microorganism |
| JP4866771B2 (en) * | 2007-03-29 | 2012-02-01 | サッポロビール株式会社 | New microorganism |
| JP4860659B2 (en) | 2008-05-12 | 2012-01-25 | シャープ株式会社 | Hydrogen generating method and hydrogen generating apparatus |
-
1990
- 1990-11-02 JP JP29538390A patent/JPH0685712B2/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| JPH04169178A (en) | 1992-06-17 |
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