JPS63309181A - Lignin-decomposition microbial strain having high activity and selectivity - Google Patents
Lignin-decomposition microbial strain having high activity and selectivityInfo
- Publication number
- JPS63309181A JPS63309181A JP14234787A JP14234787A JPS63309181A JP S63309181 A JPS63309181 A JP S63309181A JP 14234787 A JP14234787 A JP 14234787A JP 14234787 A JP14234787 A JP 14234787A JP S63309181 A JPS63309181 A JP S63309181A
- Authority
- JP
- Japan
- Prior art keywords
- lignin
- wood
- decomposition
- microbial strain
- present
- 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 18
- 230000000813 microbial effect Effects 0.000 title claims abstract description 5
- 230000000694 effects Effects 0.000 title abstract description 9
- 229920005610 lignin Polymers 0.000 claims abstract description 31
- 244000005700 microbiome Species 0.000 abstract description 22
- 238000000034 method Methods 0.000 abstract description 12
- 239000002023 wood Substances 0.000 abstract description 9
- 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
- 239000002028 Biomass Substances 0.000 abstract description 2
- 239000002994 raw material Substances 0.000 abstract description 2
- 241000894006 Bacteria Species 0.000 description 10
- 239000002609 medium Substances 0.000 description 5
- 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
- 239000000463 material Substances 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 241000123346 Chrysosporium Species 0.000 description 2
- 241000233866 Fungi 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
- 230000001580 bacterial effect Effects 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 108010059892 Cellulase Proteins 0.000 description 1
- 241000222356 Coriolus Species 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
- 241000222385 Phanerochaete Species 0.000 description 1
- 241000222393 Phanerochaete chrysosporium 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
- 238000007796 conventional method Methods 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- -1 hemicellulose Chemical class 0.000 description 1
- 239000012978 lignocellulosic material Substances 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000006395 oxidase reaction Methods 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
- 241000894007 species Species 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 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、リグニン分解能にすぐれ且つ選択的にリグニンを分
解する新規微生物に関するものである。DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to a novel microorganism, and more particularly to a novel microorganism that has excellent lignin decomposition ability and selectively decomposes lignin.
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、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、ρ５
８９〜５９０）、これらのうち、コリオラス・ヴエルシ
カラー
（Ｃｏｒｉｏｌｕｓ ｖｅｒｓｉｃｏｌｏｒ）やファネ
ロケーテ・クリソスポリウム（Ｐｈａｎｅｒｏｃｈａｅ
ｔｅ ｃｈｒｙｓｏｓｐｏｒｉｕｍ）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。(Prior art) Many people have attempted to decompose lignin using microorganisms, and it is well known that microorganisms belonging to the white-rot fungi have this activity ("Chemistry"). Great Dictionary 9”, Kyoritsu Shuppan, ρ5
89-590), among these, Coriolus versicolor and Phanerochae chrysosporium
te chrysosporium) 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
１０〜１５％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, even with the above-mentioned strain, its activity was such that only about 10 to 15% of lignin decomposition was obtained after culturing for several weeks, 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
ぐれ且つリグニン分解能の選択性にもすぐれた工業的利
用に耐え得る微生物を得るためには、既存の微生物を単
にスクリーニングするという発想を根本的に転換する必
要があるとの観点に立ち到った。そして、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
微生物を新たに分離、スクリーニングする以外に途がな
いことを痛感した。Therefore, in order to solve the above problems and obtain microorganisms that have particularly excellent lignin decomposition ability and selectivity for lignin decomposition and can withstand industrial use, we have to fundamentally change the idea of simply screening existing microorganisms. We have come to the conclusion that it is necessary to change to I realized that the only way to obtain industrially useful bacteria capable of achieving the above objectives was to newly isolate and screen previously unknown new microorganisms.
そして、目的とする新規微生物を求めて、本発明Ｍらは
、先ず、森林のほか、畑地、野草地その他より広く内外
の分離源を収集した。そしてスクリーニング法について
も、従来既知の方法のほかに先に本発明者らが発明した
ところの（特願昭６１−２８１６６８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
法も併せ用いて、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 search of the target new microorganism, the inventors M and colleagues first collected a wide range of isolation sources, including forests, fields, wild grasslands, and other sources both inside and outside the country. Regarding the screening method, in addition to conventionally known methods, the present inventors have previously invented a method (Japanese Patent Application No. 61-281668) that efficiently assays for lignin-degrading bacteria from natural samples such as decaying wood. We carried out an intensive search using these methods. 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１本発明の目的とするところの、極めてリグニ
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 an extremely high lignin decomposition activity and can selectively decompose lignin, which is the objective of the present invention, and thus arrived at the present invention.
このようにして新たに分離された微生物は、次のような
菌学的諸性質を有するものである。The microorganisms newly isolated in this way have the following mycological 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℃、４日間培養）
ｐＨ３〜９付近で生育し、ｐＨ２および１０では生育し
ない。最適ｐＨは４〜６付近である。(1) Growth conditions in the medium (2) Physiological and ecological properties■ Growth PH range (potato/glucose agar medium,
(Culture at 28°C for 4 days) It grows around pH 3 to 9 and does not grow at pH 2 and 10. The optimum pH is around 4-6.
■ 生育の温度範囲（バレイショ・ブドウ糖寒天培地、
ρ１１５．４日間培養）
１０〜４５℃付近で生育し、５０℃では生育しない。■ Temperature range for growth (potato/glucose agar medium,
ρ115.4 days culture) Grows at around 10-45°C and does not grow at 50°C.
最適温度は２８〜３７℃付近である。The optimum temperature is around 28-37°C.
■ フェノールオキシダーゼ反応（２８℃、４日間培養
）
微弱または陰性を示す。■ Phenol oxidase reaction (cultured at 28°C for 4 days) Shows weak or negative results.
■ 菌叢の特徴（バレイショ・ブドウ糖寒天培地、ＰＨ
５，２８℃、４日間培養）
白色でフェルト状である。■Characteristics of bacterial flora (potato/glucose agar medium, PH
5. Cultured at 28°C for 4 days) It is white and felt-like.
これらの生理的、生態的性質及び各培地における生育状
態といった菌学的性質を詳細に検討したが、既知の菌と
同定するには至らなかった。Although their physiological and ecological properties and mycological properties such as growth conditions in each culture medium were investigated in detail, it was not possible to identify them as known bacteria.
しかも後記する実施例からも明らかなように、リグニン
分解菌として従来より特に知られているコリオラス属菌
及びファネロケーテ属菌よりも、本菌はリグニン分解能
において特にすぐれているのみでなくその選択性につい
ても格段のものが認められ、このように卓越したリグニ
ン分解菌は従来既知の微生物には存在しないことから、
本菌株を新菌株と認定してＮＫ’−１１４８株と命名し
、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９３８４号）。本省の分類学上の正確
な位置については、なお不明な点がいくつかあるため正
確な同定が現時点においては困難であり、今後の研究に
またねばならない。Moreover, as is clear from the examples described later, this bacterium is not only superior in lignin decomposition ability, but also in its selectivity, compared to Coriolus and Phanerochaete bacteria, which have been known as lignin-degrading bacteria. This outstanding lignin-degrading bacteria does not exist among previously known microorganisms.
This strain was recognized as a new strain and named strain NK'-1148, and was deposited at the Institute of Microbial Technology, Agency of Industrial Science and Technology (Deposit number: National Institute of Microbiological Sciences No. 9384). As there are still some unknown points regarding the exact taxonomic position of this species, accurate identification is difficult at present, and will require further research.
本省を各種の木材利用工業に利用するには従来法と同様
に処理すればよく、そこには格別の配慮は何も必要とさ
れず、この点も本発明のすぐれた特徴の１つである。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
種して適温で所定時間インキニーベートすればよい。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
常に短かくてすみ、工業的に利用するのに好適である。In order to use this product in various wood-utilizing industries, it is sufficient to treat it in the same manner as conventional methods, and no special consideration is required, which is also one of the outstanding features of the present invention. . For example, in order to use this material to decompose lignin, it is necessary to inoculate a medium containing wood material with this material and incubate it at an appropriate temperature for a predetermined period of time, 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.
本発明は、また、その他木材糖化工業等にも広く利用す
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。The present invention can also be widely used in other industries such as wood saccharification, and does not require delicate operations, making it particularly suitable for industrialization. This is also one of the outstanding features of the present invention. It is one.
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
５０１容三角フラスコにブナ脱脂木粉（６０〜８０メツ
シユ） １．Ｏｇ、水２．５ｍＮを加えた培地を１２０
℃で１５分間加熱殺菌した後１本発明に係るＮＫ−１１
４８株（ＦＥＲＭ Ｐ−９３８４）、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（Ｃｏｒｉｏｌｕｓ ｖｅｒｓｉｃｏｌｏｒ）
ＩＦＯ３０３４０およびファネロケーテ・クリソスポ
リウム（Ｐｈａｎｅｒｏｃｈａｅｔｅｃｈｒｙｓｏｓｐ
ｏｒｉｕｍ） 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 mesh) in a 501 volume Erlenmeyer flask 1. 120 g of culture medium with 2.5 mN of water added.
NK-11 according to the present invention after heat sterilization at ℃ for 15 minutes
48 strain (FERM P-9384), and Coriolus versicolor, which is conventionally known as a lignin-degrading bacterium, as a control.
IFO30340 and Phanerochaetechrysosporium
orium) ATCC34541, respectively.
The cells were cultured at 28°C for one month.
培養後、木粉中のクラーソンリグニン（ＪＩＳＰ８００
８−１９６１）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
8-1961) 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。
゛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。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.
゛It can also be applied to various processes 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１本発明に係る新規微生物は、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
借越した効果を奏するものである。In addition, the novel microorganism 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, , which has the effect of being completely new and overdrafted.
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)
る微生物菌株ＮＫ−１１４８。Microbial strain NK-1148 has excellent lignin decomposition ability and selectively decomposes lignin.
Priority Applications (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP14234787A JPS63309181A (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 |
|---|---|---|---|
| JP14234787A JPS63309181A (en) | 1987-06-09 | 1987-06-09 | Lignin-decomposition microbial strain having high activity and selectivity |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS63309181A true JPS63309181A (en) | 1988-12-16 |
| JPH0332996B2 JPH0332996B2 (en) | 1991-05-15 |
Family
ID=15313252
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP14234787A Granted JPS63309181A (en) | 1987-06-09 | 1987-06-09 | Lignin-decomposition microbial strain having high activity and selectivity |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS63309181A (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 |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3262646B2 (en) * | 1993-07-09 | 2002-03-04 | 日本製紙株式会社 | Pulp bleaching method |
-
1987
- 1987-06-09 JP JP14234787A patent/JPS63309181A/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 |
|---|---|
| JPH0332996B2 (en) | 1991-05-15 |
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