JPS63309182A - Lignin-decomposition microbial strain having high activity and selectivity - Google Patents
Lignin-decomposition microbial strain having high activity and selectivityInfo
- Publication number
- JPS63309182A JPS63309182A JP14234887A JP14234887A JPS63309182A JP S63309182 A JPS63309182 A JP S63309182A JP 14234887 A JP14234887 A JP 14234887A JP 14234887 A JP14234887 A JP 14234887A JP S63309182 A JPS63309182 A JP S63309182A
- Authority
- JP
- Japan
- Prior art keywords
- lignin
- decomposition
- wood
- microbial strain
- strain
- 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.)
- Granted
Links
- 238000000354 decomposition reaction Methods 0.000 title claims abstract description 15
- 230000000694 effects Effects 0.000 title abstract description 9
- 230000000813 microbial effect Effects 0.000 title abstract description 4
- 229920005610 lignin Polymers 0.000 claims abstract description 28
- 241001061969 Porodisculus Species 0.000 claims abstract description 4
- 244000005700 microbiome Species 0.000 abstract description 20
- 238000000034 method Methods 0.000 abstract description 8
- 239000002023 wood Substances 0.000 abstract description 8
- 238000004519 manufacturing process Methods 0.000 abstract description 5
- 229920001817 Agar Polymers 0.000 abstract description 4
- 239000008272 agar Substances 0.000 abstract description 4
- 239000008103 glucose Substances 0.000 abstract description 4
- 239000002994 raw material Substances 0.000 abstract description 4
- 239000002028 Biomass Substances 0.000 abstract description 2
- 230000001580 bacterial effect Effects 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 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 3
- 244000061456 Solanum tuberosum Species 0.000 description 3
- 235000002595 Solanum tuberosum Nutrition 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 238000011282 treatment Methods 0.000 description 3
- 241000123346 Chrysosporium Species 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 241001062031 Porodisculus pendulus Species 0.000 description 2
- 229920001131 Pulp (paper) Polymers 0.000 description 2
- 241000222355 Trametes versicolor Species 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 108010059892 Cellulase Proteins 0.000 description 1
- 240000000731 Fagus sylvatica Species 0.000 description 1
- 235000010099 Fagus sylvatica Nutrition 0.000 description 1
- 229920002488 Hemicellulose Polymers 0.000 description 1
- 241000391444 Oriolus Species 0.000 description 1
- 241000222393 Phanerochaete chrysosporium Species 0.000 description 1
- 241000195888 Physcomitrella Species 0.000 description 1
- 102000003425 Tyrosinase Human genes 0.000 description 1
- 108060008724 Tyrosinase Proteins 0.000 description 1
- 229940106157 cellulase Drugs 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 210000004209 hair Anatomy 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- -1 hemicellulose Chemical class 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000012978 lignocellulosic material Substances 0.000 description 1
- 238000006395 oxidase reaction Methods 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 238000004076 pulp bleaching Methods 0.000 description 1
- 238000004537 pulping Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 235000013580 sausages Nutrition 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004065 wastewater treatment Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
【発明の詳細な説明】
（産業上の利用分野）
本発明は、新規微生物に関するものであり、更に詳細に
は、リグニン分解能に優れ且つ選択的にリグニンを分解
する性質を有するヌルデタケに属する新規菌株Ｐｏｒｏ
ｄｉｓｃｕｌｕｓ ｐｅｎｄｕｌｕｓ Ｎに一７２９ｖ
に関するものである。Detailed Description of the Invention (Industrial Application Field) The present invention relates to a novel microorganism, and more specifically, a novel strain belonging to the Physcomitrella sp. Poro
disculus pendulus N 1729v
It is related to.
したがって本発明は、木材糖化、紙パルプ製造等といっ
た各種木材利用工業の技術分野をはじめとして、その他
リグニンを分解及び／又は低分子化させるすべての技術
分野において広く利用されるものである。Therefore, the present invention can be widely used in technical fields of various wood utilization industries such as wood saccharification and paper pulp production, as well as all other technical fields in which lignin is decomposed and/or reduced in molecular weight.
（従来の技術）
従来より、リグニンを微生物によって分解しようとする
試みは多くの人々によってなされており。(Prior Art) Many people have attempted to decompose lignin using microorganisms.
白色腐朽菌に属する微生物にその活性が見られることは
よく知られているところである（「化学大辞典９」、共
立出版、ｐ５８９〜５９０）。これらのうち。It is well known that this activity is observed in microorganisms belonging to the white rot fungi ("Chemistry Dictionary 9", Kyoritsu Shuppan, p. 589-590). Of these.
コリオラス・ヴエルシカラー
（Ｃｏｒｉｏｌｕｓ ｖｅｒｓｉｃｏｌｏｒ）やファネ
ロケーテφクリソスポリウム（Ｐｈａｎｅｒｏｃｈａｅ
ｔｅ ｃｈｒｙｓｏｓｐｏｒｉｕ＋ａ）などが代表的な
微生物として注目され、主として研究開発の目的で用い
られてきた。Coriolus versicolor and Phanerochae φ Chrysosporium
te chrysosporiu+a) and the like have attracted attention as representative microorganisms and have been used mainly for research and development purposes.
しかしながら、上記菌株でもその活性は数週間の培養で
ｌＯ〜１５％程度のリグニン分解が得られるのみで、余
りに活性が低いため実用化には至らなかった。つまり、
従来既知の菌株では工業的処理を行うことはできなかっ
たのである。However, the activity of the above-mentioned bacterial strain was such that only about 15% of lignin was degraded after several weeks of culture, and the activity was so low that it could not be put to practical use. In other words,
It was not possible to carry out industrial treatment with conventionally known bacterial strains.
（発明が解決しようとする問題点）
このように、既存の白色腐朽菌ではリグニン分解に長期
間を要するうえに１分解力が極めて不充分であり、さら
にはリグニン分解と同時に、共存するセルロースやヘミ
セルロース等の炭水化物をも分解するため、木材糖化等
の有用な工業的用途に利用することは不可能であった。(Problems to be solved by the invention) As described above, existing white rot fungi require a long period of time to decompose lignin, and their decomposition power per unit is extremely insufficient. Because it also decomposes carbohydrates such as hemicellulose, it has been impossible to use it for useful industrial purposes such as wood saccharification.
工業的用途に使用し得る微生物としては、リグニン分解
力が非常にすぐれ、しかもそのうえ選択的にリグニンを
分解する性質をも併有するものでなければならないが、
従来、このような微生物は知られていなかったのである
。Microorganisms that can be used for industrial purposes must have extremely high lignin decomposition ability and also have the ability to selectively decompose lignin.
Until now, such microorganisms were unknown.
（問題点を解決するための手段）
本発明は、上記した技術の現状に鑑みてなされたもので
あって、既存の微生物について各種のスクリーニングを
くり返したけれども目的とする微生物を得るには到らな
かった。(Means for Solving the Problems) The present invention has been made in view of the current state of the technology described above, and although various screenings have been repeated for existing microorganisms, it has not been possible to obtain the target microorganisms. There wasn't.
そこで、上記問題点を解決して特にリグニン分解力にす
ぐれ且つ選択的にリグニンを分解しうる工業的利用に耐
えうる微生物を得るためには、既存の微生物に頼ったの
では所期の目的が達成されないとの観点に立ち到った。Therefore, in order to solve the above problems and obtain a microorganism that can withstand industrial use and has particularly excellent lignin decomposition ability and can selectively decompose lignin, relying on existing microorganisms would not meet the intended purpose. I came to the point that it would not be achieved.
そこで、上記目的を達成しうる工業的に有用な菌を得る
には、従来未知の新規微生物をスクリーニングする必要
性を痛感したのである。Therefore, in order to obtain industrially useful microorganisms that can achieve the above objectives, we became acutely aware of the necessity of screening previously unknown new microorganisms.
そして、目的とする新規微生物を得るため、先ず、本発
明者らは、広く内外の森林、畑地、野草地その他より分
離源を収集した。そしてスクリーニング方法にも工夫を
こらし、先に本発明者等が発明したところの（特願昭６
１−２８１６６８号）、腐朽材など自然界の試料から効
率的にリグニン分解菌を検定する方法により、鋭意検索
を行なった。即ち、膨大な分離源と新検定法との組合せ
により強力なリグニン分解菌の検索分離を行なったので
ある。In order to obtain the desired new microorganism, the present inventors first collected isolated sources from forests, fields, wild grasslands, and other areas in Japan and abroad. We also devised a screening method, which was invented by the present inventors (patent application filed in 1983).
No. 1-281668), we carried out an intensive search using a method that efficiently assays lignin-degrading bacteria from natural samples such as decayed wood. In other words, we were able to search for and isolate powerful lignin-degrading bacteria by combining a huge number of isolation sources and a new assay method.
その結果、本発明の目的とするところの、Ｖｉめてリグ
ニン分解活性が高く、かつリグニンを選択的に分解しう
る新規微生物を分離することに成功し、本発明に到達し
たのである、。As a result, we succeeded in isolating a new microorganism that has a high lignin decomposition activity and can selectively decompose lignin, which is the objective of the present invention, and the present invention has been achieved.
このようにして新たに分離された微生物は、次のような
生育状態、生理学、生態学的性質を有するものである。The microorganisms newly isolated in this manner have the following growth conditions, physiology, and ecological properties.
（１）培地における生育状態
（２）生理的、生態的性質
■ 生育のｐＨ範囲（バレイショ・ブドウ糖寒天培地、
２８℃、培養４日間）
ＰＨ３〜７付近で生育し、ｐＨ２および８では生育しな
い。最適ｐＨは４〜５付近である。(1) Growth conditions in the medium (2) Physiological and ecological properties■ pH range for growth (potato/glucose agar medium,
28°C, 4 days of culture) Grows around pH 3 to 7, and does not grow at pH 2 and 8. The optimum pH is around 4-5.
■ 生育の温度範囲（バレイショ・ブドウ糖寒天培地、
ｐＨ５、培養４日間）
１０〜３２℃付近で生育し、３７℃では生育しない。■ Temperature range for growth (potato/glucose agar medium,
(pH 5, culture for 4 days) It grows at around 10-32°C and does not grow at 37°C.
最適温度は２０〜３０℃付近である。The optimum temperature is around 20-30°C.
■ フェノールオキシダーゼ反応（２８℃、培養４日間
）陽性を示す。■ Phenol oxidase reaction (28°C, cultured for 4 days) is positive.
■ 菌叢の形状（バレイショ・ブドウ糖寒天培地、ｐＨ
５，２８℃、培養４日間）
白色で毛状である。■ Shape of bacterial flora (potato/glucose agar medium, pH
5.28°C, 4 days of culture) White and hairy.
■ 子実体の形状
・大きさ ：径２〜５ＩＩＩ■
・形 ：倒椀形（鼻の形）
・縁や表面 ：縁は内側へ巻き、表面は黄褐色で全面に
褐色毛がある６
・管孔面 ：淡灰白色で倒皿状にへこんでおり管孔は
小さい。■ Shape and size of fruiting body: Diameter 2-5III■ ・Shape: Inverted bowl shape (nose shape) ・Edge and surface: Edge curls inward, surface is yellowish brown with brown hairs all over 6 ・Tube Pore surface: Pale grayish white, concave in the shape of an inverted dish, and the pores are small.
・肉質 ：柔軟な革質でほぼ白色である。・Flesh: Flexible, leathery and almost white in color.
■ 胞子の形状 ３〜４×１μｍ程度のソーセージ形で無色平滑である。■ Spore shape It is colorless and smooth with a sausage shape of about 3 to 4 x 1 μm.
これらの菌学的性質からして、本省は、ポロディスキュ
ラス属に属するものと認められ、分類学上、特にその内
のヌルデタケ（Ｐｏｒｏｄｉｓｃｕｌｕｓｐｅｎｄｕｌ
ｕｓ）に属せしめるのが適当であると認められた。Based on these mycological properties, the Ministry recognizes that they belong to the genus Porodisculus, and taxonomically, they are particularly sensitive to the genus Porodisculus.
It was recognized that it was appropriate to make it belong to the United States.
しかしながら、後記する実施例からも明らかなように、
本省は卓越したリグニン分解能を示すのみでなくその選
択性も格段にすぐれており、このような卓越した性質を
示す菌は従来既知のヌルデタケには存在しないことから
、本省はヌルデタケの新菌株と認定し、これをボロディ
スキュラス・ペンデュラスＮＫ−７２９すと命名して工
業技術院微生物工業技術研究所に寄託した（寄託番号：
微工研菌寄第９３８５号）。However, as is clear from the examples described later,
This strain not only exhibits excellent lignin decomposition ability but also extremely high selectivity, and as there are no previously known strains of Nurudetake that exhibit such outstanding properties, the Ministry has recognized it as a new strain of Nurudetake. This was named Borodisculus pendulus NK-729 and deposited at the Institute of Microbial Technology, Agency of Industrial Science and Technology (Deposit number:
Microtechnical Research Institute No. 9385).
本省を例えばリグニンの分解に利用するには、他のリグ
ニン分解菌の場合と同様に、木質原料含有培地に本省を
接種して適温でインキュベートすればよい。本省はその
性能が卓越しているために、大型タンクを用いる大量処
理、大量生産に特に適しており、処理時間も非常に短か
く、工業的に利用するのに好適である。また、木材糖化
工業やその他の処理にも１本省は自由に使用することが
でき、しかもその際デリケートな操作を必要としないの
で、工業化には特に適しており、この点も本発明のすぐ
れた特徴の１つである。For example, in order to use this species to decompose lignin, it is sufficient to inoculate a medium containing woody raw materials with this species and incubate it at an appropriate temperature, as in the case of other lignin-degrading bacteria. Due to its outstanding performance, this product is particularly suitable for mass processing and mass production using large tanks, and the processing time is very short, making it suitable for industrial use. In addition, it can be freely used in the wood saccharification industry and other treatments, and does not require delicate operations, making it particularly suitable for industrialization, and this is another advantage of the present invention. This is one of its characteristics.
次に本発明の実施例を示す。Next, examples of the present invention will be shown.
実施例１
５０ｍｆｌ容三角フラスコにブナ脱脂木粉（６０〜８０
メツシユ） １．０ｇ、水２．５＋Ｉ２を加えた培地を
１２０℃で１５分間加熱殺菌した後、Ｐｏｒｏｄｉｓｃ
ｕｌｕｓ ｐｅｎｄｕｌｕｓＮＫ−７２９Ｗ株、（ＦＥ
ＲＭ Ｐ−９３８５）、並びに、対照としてリグニン分
解菌として従来既知のコリオラス・ヴエルシカラー（Ｃ
ｏｒｉｏｌｕｓ ｖｅｒｓｉｃｏｌｏｒ）ＩＦＯ３０３
４０およびファネロケーテ・クリソスポリウム（Ｐｈａ
ｎｅｒｏｃｈａｅｔｅ ｃｈｒｙｓｏｓｐｏｒｉｕｍ）
ＡＴＣＣ３４５４１をそれぞれ接種し、２８℃で１ケ月
間培養した。Example 1 Beech defatted wood flour (60-80%
After heating and sterilizing a medium containing 1.0 g of methane and 2.5 g of water + I2 at 120°C for 15 minutes, Porodisc
ulus pendulus NK-729W strain, (FE
RM P-9385), and Coriolus versicolor (C
oriolus versicolor) IFO303
40 and Phanerochaete chrysosporium (Pha
chrysosporium)
ATCC34541 was inoculated and cultured at 28°C for one month.
培養後、木粉中のクラーソンリグニン（ＪＩＳＰ８００
ｇ−１９６１）および酸可溶性リグニン（中野準三編：
リグニンの化学、ｐ５３（１９８２））を定量した。な
お、重量減少も測定し、炭水化物量を算出した。After culturing, Klason lignin (JISP800
g-1961) and acid-soluble lignin (edited by Junzo Nakano:
Chemistry of Lignin, p53 (1982)) was quantified. In addition, weight loss was also measured and the amount of carbohydrates was calculated.
その結果を表１に示す。The results are shown in Table 1.
上記表１の結果からも明らかなように、本発明に係る新
規微生物はリグニン分解能が格段にすぐれ、選択性も非
常に高いことが判る。As is clear from the results in Table 1 above, the novel microorganism according to the present invention has an extremely excellent ability to decompose lignin and has very high selectivity.
（発明の効果）
本発明は、リグニン分解能に優れ、かつ選択的にリグニ
ンを分解する新規微生物の開発に成功したものである。(Effects of the Invention) The present invention has successfully developed a novel microorganism that has excellent lignin decomposition ability and selectively decomposes lignin.
したがって１本発明に係る微生物はリグニンに作用して
これを分解または低分子化するすぐれた性質を有するた
め、木材などのリグノセルロース物質を原料とする糖化
工程において、糖化の前段処理としてリグニンを分解す
ることによってセルラーゼの作用を高めるというセルロ
ース系バイオマス利用の分野に有利に利用できる。Therefore, the microorganism according to the present invention has an excellent property of acting on lignin and decomposing it or reducing its molecular weight. Therefore, in the saccharification process using lignocellulosic materials such as wood as a raw material, it decomposes lignin as a preliminary treatment of saccharification. By doing so, it can be advantageously used in the field of cellulosic biomass utilization to enhance the action of cellulase.
また、紙パルプ製造工程における種々の工程にも適用で
きる。すなわち、パルプ化工程、パルプ漂白工程、排水
処理工程等におけるリグニンの分解または低分子化を行
なわせることにも有利に利用できる。It can also be applied to various steps in the paper pulp manufacturing process. That is, it can be advantageously used to decompose or reduce the molecular weight of lignin in pulping processes, pulp bleaching processes, wastewater treatment processes, and the like.
なお、その他リグニンを分解または低分子化させる技術
分野に全て適用できる。In addition, it can be applied to all other technical fields in which lignin is decomposed or reduced in molecular weight.
しかも、本発明に係る新菌株は、リグニンの分解能及び
選択性がきわめて高いので、いずれの用途においても工
業的大量処理に有利に利用することができるものであっ
て、従来既知の微生物とは異なり、全く新規にして卓越
した効果を奏するものである。Moreover, the new bacterial strain according to the present invention has extremely high lignin decomposition ability and selectivity, so it can be advantageously used for industrial large-scale processing in any application, and unlike conventionally known microorganisms, it can be used advantageously for industrial large-scale processing. , which is completely new and has outstanding effects.
このように本発明によって、はじめて、リグニン分解の
工業化が可能となったのである。In this way, the present invention has made it possible for the first time to industrialize lignin decomposition.
Claims (1)
ｕｓ）に属し、リグニン分解能に優れ、且つ選択的にリ
グニンを分解する菌株ＮＫ−７２９Ｗ。Porodisculus pendul
strain NK-729W, which belongs to the US) and has excellent lignin decomposition ability and selectively decomposes lignin.
Priority Applications (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP14234887A JPS63309182A (en) | 1987-06-09 | 1987-06-09 | Lignin-decomposition microbial strain having high activity and selectivity |
| CA000568230A CA1321158C (en) | 1987-06-09 | 1988-05-31 | Lignin decomposing bacteria having high activity and selectivity |
| EP88305196A EP0295063B1 (en) | 1987-06-09 | 1988-06-08 | Lignin-degrading microorganism having high activity and selectivity |
| FI882743A FI88406C (en) | 1987-06-09 | 1988-06-09 | Extremely active and selective microorganisms that decompose lignin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP14234887A JPS63309182A (en) | 1987-06-09 | 1987-06-09 | Lignin-decomposition microbial strain having high activity and selectivity |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS63309182A true JPS63309182A (en) | 1988-12-16 |
| JPH0332997B2 JPH0332997B2 (en) | 1991-05-15 |
Family
ID=15313279
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP14234887A Granted JPS63309182A (en) | 1987-06-09 | 1987-06-09 | Lignin-decomposition microbial strain having high activity and selectivity |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS63309182A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1995004131A1 (en) * | 1993-07-28 | 1995-02-09 | Nippon Seishi Kabushiki Kaisha | Microbe skb-1152 strain, method of separating the same, and method of bleaching pulp therewith |
-
1987
- 1987-06-09 JP JP14234887A patent/JPS63309182A/en active Granted
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1995004131A1 (en) * | 1993-07-28 | 1995-02-09 | Nippon Seishi Kabushiki Kaisha | Microbe skb-1152 strain, method of separating the same, and method of bleaching pulp therewith |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0332997B2 (en) | 1991-05-15 |
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