JPS6391077A - Inoculation of bacterium into lignocellulose - Google Patents
Inoculation of bacterium into lignocelluloseInfo
- Publication number
- JPS6391077A JPS6391077A JP23722386A JP23722386A JPS6391077A JP S6391077 A JPS6391077 A JP S6391077A JP 23722386 A JP23722386 A JP 23722386A JP 23722386 A JP23722386 A JP 23722386A JP S6391077 A JPS6391077 A JP S6391077A
- Authority
- JP
- Japan
- Prior art keywords
- culture medium
- microorganisms
- suspension
- valve
- inoculation
- 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
- 238000011081 inoculation Methods 0.000 title claims description 15
- 241000894006 Bacteria Species 0.000 title description 7
- 238000000034 method Methods 0.000 claims abstract description 23
- 239000000725 suspension Substances 0.000 claims abstract description 19
- 229920005610 lignin Polymers 0.000 claims abstract description 12
- 241000221198 Basidiomycota Species 0.000 claims abstract description 4
- 244000005700 microbiome Species 0.000 claims description 33
- 239000012978 lignocellulosic material Substances 0.000 claims description 20
- 239000007788 liquid Substances 0.000 claims description 8
- 235000015097 nutrients Nutrition 0.000 claims description 5
- 230000001954 sterilising effect Effects 0.000 claims description 5
- 241000123346 Chrysosporium Species 0.000 claims description 3
- 239000000835 fiber Substances 0.000 claims description 2
- 241000233866 Fungi Species 0.000 abstract description 16
- 238000001816 cooling Methods 0.000 abstract description 9
- 239000000243 solution Substances 0.000 abstract description 9
- 239000002023 wood Substances 0.000 abstract description 8
- 239000000126 substance Substances 0.000 abstract description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 abstract description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 3
- 241000222393 Phanerochaete chrysosporium Species 0.000 abstract description 2
- 244000052616 bacterial pathogen Species 0.000 abstract description 2
- 238000010438 heat treatment Methods 0.000 abstract description 2
- 239000011780 sodium chloride Substances 0.000 abstract description 2
- 239000001963 growth medium Substances 0.000 abstract 5
- 244000176261 Viburnum cassinoides Species 0.000 abstract 1
- 239000008354 sodium chloride injection Substances 0.000 abstract 1
- 238000011109 contamination Methods 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 239000002994 raw material Substances 0.000 description 5
- 230000000694 effects Effects 0.000 description 4
- 239000002504 physiological saline solution Substances 0.000 description 4
- 238000004537 pulping Methods 0.000 description 4
- 238000012258 culturing Methods 0.000 description 3
- 239000002054 inoculum Substances 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- 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 2
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 2
- 241000222385 Phanerochaete Species 0.000 description 2
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 description 2
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000000428 dust Substances 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000010025 steaming Methods 0.000 description 2
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 1
- 229930024421 Adenine Natural products 0.000 description 1
- 239000004254 Ammonium phosphate Substances 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- AUNGANRZJHBGPY-UHFFFAOYSA-N D-Lyxoflavin Natural products OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-UHFFFAOYSA-N 0.000 description 1
- 229920002488 Hemicellulose Polymers 0.000 description 1
- 101000876606 Homo sapiens Putative uncharacterized protein encoded by ERC2-IT1 Proteins 0.000 description 1
- 102100035223 Putative uncharacterized protein encoded by ERC2-IT1 Human genes 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- JZRWCGZRTZMZEH-UHFFFAOYSA-N Thiamine Natural products CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N JZRWCGZRTZMZEH-UHFFFAOYSA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 229960000643 adenine Drugs 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 229910000148 ammonium phosphate Inorganic materials 0.000 description 1
- 235000019289 ammonium phosphates Nutrition 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 235000013877 carbamide Nutrition 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 235000008216 herbs Nutrition 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 229960002477 riboflavin Drugs 0.000 description 1
- 235000019192 riboflavin Nutrition 0.000 description 1
- 239000002151 riboflavin Substances 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- -1 shakerose Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 235000019157 thiamine Nutrition 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- 239000011721 thiamine Substances 0.000 description 1
- KYMBYSLLVAOCFI-UHFFFAOYSA-N thiamine Chemical compound CC1=C(CCO)SCN1CC1=CN=C(C)N=C1N KYMBYSLLVAOCFI-UHFFFAOYSA-N 0.000 description 1
- 229960000344 thiamine hydrochloride Drugs 0.000 description 1
- 235000019190 thiamine hydrochloride Nutrition 0.000 description 1
- 239000011747 thiamine hydrochloride Substances 0.000 description 1
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 1
- 229940035893 uracil Drugs 0.000 description 1
- 238000002255 vaccination Methods 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Paper (AREA)
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明は、リグノセルロース物質にリグニン分解能を有
する微生物を接種する方法に関するものである。さらに
詳しくは、リグニン分解能を有する微生物を利用してパ
ルプを製造するいわゆるバイオパルピングにおいて、リ
グノセルロース物質にリグニン分解能を有する微生物、
特に白色腐朽菌を、雑菌に汚染されることなく、効率よ
く接種する方法に関するものである。DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a method of inoculating lignocellulosic material with microorganisms capable of degrading lignin. More specifically, in so-called biopulping, in which pulp is produced using microorganisms that have the ability to decompose lignin, microorganisms that have the ability to decompose lignin,
In particular, the present invention relates to a method for efficiently inoculating white rot fungi without being contaminated with germs.
自然界でリグニンを分解すると考えられている微生物は
現在多数報告されている。その中で白色腐朽菌と呼ばれ
ている１群の菌類は、木材等のリグノセルロース物質中
のリグニンを分解するｔＵＪが高いことが知られている
。この白色腐朽菌をリグノセルロース物質に接種、培養
し、リグニンの一部を分解させてパルプを製造するバイ
オパルピングの試みがなされている（特開昭５０−４６
９０３号）。Many microorganisms that are thought to decompose lignin in nature have now been reported. Among them, one group of fungi called white rot fungi is known to have a high tUJ that decomposes lignin in lignocellulose materials such as wood. Attempts have been made to produce pulp by inoculating and culturing this white-rotting fungus into lignocellulosic materials and decomposing part of the lignin (Japanese Patent Laid-Open No. 50-46
No. 903).
リグノセルロース物質、殊に木材に白色腐朽菌を接種す
る一般的方法は、キノコの人工培養にみられるように、
菌糸の蔓延した種駒を猾木、ソー・ダスト等に埋め込む
方法である。この方法は、極めて局所的な接種方法であ
り、数ケ月以上に亙る培養期間中に菌糸が木材中に伸び
、最終的に木材全体に蔓延させる方法である。A common method of inoculating lignocellulosic materials, especially wood, with white-rot fungi is to
This is a method of embedding seed pieces infested with mycelium in wood, saw dust, etc. This method is a very localized inoculation method in which mycelia grow into the wood during a cultivation period that lasts several months or more, eventually spreading throughout the wood.
しかし、白色腐朽菌によるリグニン分解をパルプ化に利
用した大規模なバイオパルピングを考えた場合には培養
中の他の微生物の混入の防止や製造コストの低下のため
に２週間以内の短期間の培養が望ましい。したがって、
従来のように局所的な菌糸の伸長だけに期待した微生物
の接種法ではバイオパルピングを行なうのは難しい。However, when considering large-scale biopulping that utilizes lignin decomposition by white rot fungi for pulping, it is necessary to carry out a short period of less than two weeks to prevent contamination with other microorganisms during culture and to reduce production costs. culture is desirable. therefore,
It is difficult to perform biopulping using the conventional inoculation method of microorganisms, which only aims at localized hyphal elongation.
また、ソーダストチップダストの如き細片化したリグノ
セルロース物質に微生物を接種する際には、雑菌の混入
を防止するために原料を蒸気滅菌することが一般的であ
る。この場合原料の蒸気滅菌後は、接種する微生物の培
養に適した温度まで冷却する工程が必要であり、リグニ
ンを分解することが知られている白色腐朽菌を接種する
場合、菌糸より耐熱性があるとされている胞子でさえ５
０℃以上の温度に曝されると経時的に死滅していくため
、通常は原料を冷却した後に接種し、菌の接種の安定性
、均一性を保っている。したがって、大規模なバイオパ
ルピングを考えた場合もスチーミングやオートクレーブ
等の蒸気滅菌が考えられるが、この後に接種する微生物
の培養に適した温度まで下げる冷却工程にかなりのコス
トがかかると予想される。また、冷却工程に時間がかか
り、微生物の接種、培養までの間に雑菌による汚染の可
能性も考えられる。Furthermore, when inoculating microorganisms into fragmented lignocellulosic material such as sawdust chip dust, the raw material is generally steam sterilized to prevent contamination with bacteria. In this case, after steam sterilization of the raw material, a step of cooling to a temperature suitable for culturing the microorganisms to be inoculated is necessary. Even the spores that are said to be 5
If exposed to temperatures above 0°C, they will die over time, so the inoculation is usually done after the raw material has been cooled to maintain stability and uniformity of the inoculation. Therefore, when considering large-scale biopulping, steam sterilization such as steaming or autoclaving may be considered, but the subsequent cooling process to lower the temperature to a temperature suitable for culturing the microorganisms to be inoculated is expected to be quite costly. Ru. In addition, the cooling process takes time, and there is a possibility of contamination with various bacteria during the period from inoculation and cultivation of microorganisms.
本発明は、前述の従来の問題点を解消し、リグノセルロ
ース物質に微生物を均一に接種し、微生物のリグノセル
ロース物質全体への蔓延を約２週間で行なえるようにし
て雑菌の混入を防ぐとともにバイオパルピングを効率的
に行なうことを目的とするものである。The present invention solves the above-mentioned conventional problems, uniformly inoculates the lignocellulosic material with microorganisms, allows the microorganisms to spread throughout the lignocellulosic material in about two weeks, and prevents the contamination of various bacteria. The purpose is to perform biopulping efficiently.
さらに本発明は、冷却工程を省略することにより、製造
コストを低減することを目的とするものである。Furthermore, the present invention aims to reduce manufacturing costs by omitting the cooling step.
本発明はリグノセルロース物質に微生物を作用させ、リ
グニンを分解する方法において、リグノセルロース物質
を加熱滅菌し、次いで該リグノセルロース物質に微生物
の懸濁液を高温下で接触せしめることを特徴とする微生
物のリグノセルロース物質への接種方法である。The present invention provides a method for decomposing lignin by allowing a microorganism to act on a lignocellulosic material, the method comprising heating the lignocellulosic material, and then contacting the lignocellulosic material with a suspension of the microorganism at a high temperature. This is a method of inoculating lignocellulosic materials.
一般にリグノセルロース物質を加熱滅菌した後で微生物
、特に白色腐朽菌を接種する場合、冷却することなしに
接種の処理を行うと、白色腐朽菌は、高温に曝され死滅
し接種が極めて不良乃至不可能となる。したがって通常
は、冷却を経た後に接種する。Generally, when inoculating microorganisms, especially white rot fungi, after heat sterilizing lignocellulosic materials, if the inoculation is performed without cooling, the white rot fungi will be exposed to high temperatures and die, resulting in extremely poor or incomplete inoculation. It becomes possible. Therefore, inoculation is usually performed after cooling.
しかし、本発明は冷却することなしに高温下で微生物、
特に白色腐朽菌の分生胞子を）キ濁液としてリグノセル
ロース物質に接触させることにより菌の接種を良好に行
うことを可能にした。即ち、微生物を懸、濁液として接
触させることから、送入された胞子懸濁液の先端は高温
になるが、後続する微生物懸濁液が曝される温度は、次
第に低下する。その上チップの層があるため液の移動が
阻止され、液の対流による熱の移動が少ない。このため
、送入された微生物懸濁液は、必然的に温度差を生じ、
懸濁液中の全ての微生物が高温に曝されることにならな
いため、菌の接種が可能となる。However, the present invention allows microorganisms to be extracted at high temperatures without cooling.
In particular, by bringing conidia of white-rot fungi into contact with lignocellulose materials as a suspension, it has become possible to successfully inoculate the fungi. That is, since the microorganisms are suspended and brought into contact as a turbid liquid, the tip of the introduced spore suspension becomes high in temperature, but the temperature to which the subsequent microorganism suspension is exposed gradually decreases. Furthermore, the layer of chips prevents the movement of liquid, and there is less heat transfer due to liquid convection. For this reason, the introduced microbial suspension inevitably experiences a temperature difference,
Bacterial inoculation is possible because all the microorganisms in the suspension are not exposed to high temperatures.
微生物を接種し、培養器内の温度が平衡化した後に胞子
を懸濁していた溶液を排出することにより、大部分の熱
を排出することができ、世子閑の培養に適した温度、か
つ固体培養の状態となり、そのまま菌を培養することが
できる。By inoculating the microorganisms and discharging the solution in which the spores were suspended after the temperature in the incubator has equilibrated, most of the heat can be discharged, and the temperature is suitable for cultivating Seikokan. The cell enters a culture state and the bacteria can be cultured as is.
リグセルロース物質に微生物、特に白色腐朽菌を接種す
る種菌の形態としては、胞子と菌糸がある。液体培養の
場合には、種菌の形態は接種の均一性に大きな影響を与
えることはないがリグノセルロース物質は固体であるた
め、糸状の菌糸を用いると原料表面に堆積されてしまい
、均一な閑の接種が困難となる。この点、胞子は形状が
小さいので胞子Ｑｉ液を用いることにより（原料表面に
堆積されることなく）均一な接種を行うことができる。The forms of inoculum used to inoculate microorganisms, particularly white rot fungi, into ligcellulose materials include spores and hyphae. In the case of liquid culture, the morphology of the inoculum does not have a major effect on the uniformity of inoculation, but since lignocellulosic material is solid, if filamentous mycelium is used, it will be deposited on the surface of the raw material, resulting in a uniform inoculum. This makes vaccination difficult. In this regard, since spores are small in shape, uniform inoculation can be performed by using the spore Qi solution (without being deposited on the surface of the raw material).
さて、白色腐朽菌の多くが属する担子菌には、有性胞子
である担子胞子と分生胞子等の無性胞子がある。菌の接
種には、いずれの胞子を利用しても良いが、胞子の調製
は分生胞子の方が前便である場合が多い。Basidiomycetes, to which most white rot fungi belong, include sexual spores, basidiospores, and asexual spores, such as conidia. Although any spores may be used for inoculating the fungus, spores are often prepared using conidia.
本発明に用いるリグノセルロース物質とは、針葉樹、広
葉樹、草本等の植物体の主要構成成分中、リグニンを含
むセルロース、ヘミセルロース等のことでその形状がチ
ップ状あるいは繊維状のものである。The lignocellulose material used in the present invention refers to cellulose, hemicellulose, etc. containing lignin, which is one of the main constituents of plants such as coniferous trees, broad-leaved trees, and herbs, and is in the form of chips or fibers.
本発明に用いることのできる微生物は、リグニン分解能
があり、胞子を形成するものであれば全て可能であるが
、特に脱リグニン能力が大きく、生長の早い担子菌のフ
ァネロケーテ・クリソスポリウム（Ｐｈａｎｅｒｏｃｈ
ａｅｔｅ ｃｈｒｙｓｏｓｐｏｒｉｕｍ （八ＴＣＣ２
４７２５）、及びＰｈａｎｅｒｏｃｈａｅｔｅ ｃｈｒ
ｙｓｏｓｐｏｒｉｕｍ （八ＴＣＣ３４５４１） ）が
存効である。Any microorganism that can be used in the present invention can be used as long as it has the ability to decompose lignin and form spores. In particular, the basidiomycete Phanerochaete chrysosporium has a large delignification ability and is a fast-growing microorganism.
aete chrysosporium (8TCC2
4725), and Phanerochaete chr.
ysosporium (8TCC34541)) is still active.
又、本発明に用いる胞子懸濁液は、担子菌分生胞子を生
理食塩水に懸濁させてもよいが、その微生物の生育を促
進させるグルコース、シェークロース、デンプンの加水
分解物、廃糖蜜等の炭素源、硫安、リン酸アンモニウム
、塩化アンモニウム、尿素、アミノ酸等の窒素源、ビオ
チン、チアミン、リボフラビン、ニコチン酸等のビタミ
ン類、アデニン、グアニン、ウラシル等の核酸類を含む
溶液中に懸濁させても何ら問題はない。In addition, the spore suspension used in the present invention may be made by suspending basidiomycete conidia in physiological saline, but glucose, shakerose, starch hydrolyzate, and molasses that promote the growth of the microorganisms may be used. Suspended in a solution containing carbon sources such as ammonium sulfate, ammonium phosphate, ammonium chloride, urea, and amino acids, vitamins such as biotin, thiamine, riboflavin, and nicotinic acid, and nucleic acids such as adenine, guanine, and uracil. There is no problem in making it muddy.
以下、本発明を実施例によって説明するが本発明は、こ
れらの実施例に限定されるものではない。EXAMPLES The present invention will be described below with reference to Examples, but the present invention is not limited to these Examples.
実施例１
第１図は本願発明を実施するための装置の略図であって
、外部から胞子懸濁液２を流入するためのバイブ３を持
つオートクレーブ８に、圧力弁１０を持つ２１容量の培
養器を接続し、乾燥重量１５０ｇの木片を入れ、液比１
になるように水を加え、１２１℃、３０分間滅菌処理を
行う。このとき、圧力弁１０、バルブ４．５は開放して
おく。Example 1 FIG. 1 is a schematic diagram of an apparatus for carrying out the present invention, in which an autoclave 8 with a vibrator 3 for injecting a spore suspension 2 from the outside, and a 21-volume culture culture vessel with a pressure valve 10 are provided. Connect the container, add a piece of wood with a dry weight of 150g, and add a liquid ratio of 1.
Add water to the solution and sterilize at 121°C for 30 minutes. At this time, pressure valve 10 and valve 4.5 are kept open.
通気用の無菌フィルター１２の付いた胞子懸濁液タンク
１には、２×１０６個胞子／ｆｆ１／の無菌生理食塩水
の胞子Ｑ ？ｒＪ液２を準備する。滅菌後、圧力弁９よ
り圧力を抜き、その後に圧力弁９は閉じる。A spore suspension tank 1 equipped with a sterile filter 12 for ventilation contains 2 x 106 spores/ff1/ of sterile saline spores Q? Prepare rJ solution 2. After sterilization, the pressure is released from the pressure valve 9, and then the pressure valve 9 is closed.
次に閉じていたバルブ６．７を開放すると胞子懸濁液が
流入しはじめ、同時にオートクレーブ内が冷され、減圧
状態となり、さらに流入が進む。Next, when the previously closed valve 6.7 is opened, the spore suspension begins to flow in. At the same time, the inside of the autoclave is cooled and the pressure is reduced, and the flow continues further.
液比が４、すなわち４５０ｍｆの胞子懸濁液が流入した
ら、バルブ６を閉じ、次に圧力弁９を開放し、常圧に戻
す。オートクレーブのフタを開け、バルブ４を閉め、培
養器１１を取り出し、圧力弁１０で圧力を調整した後に
、バルブ４から胞子を懸濁させていた生理食塩水を排水
する。この時点で、内部温度は４５°Ｃとなる。When the spore suspension with a liquid ratio of 4, ie, 450 mf, flows in, the valve 6 is closed, and the pressure valve 9 is then opened to return to normal pressure. The lid of the autoclave is opened, the valve 4 is closed, the incubator 11 is taken out, and the pressure is adjusted with the pressure valve 10, and then the physiological saline in which the spores are suspended is drained from the valve 4. At this point, the internal temperature is 45°C.
培養器を３７℃に移し、培養した。この方法によりＰｈ
ａｎｅｒｏｃｈａｅｔｅ ｃｈｒｙｓｏｓｐｏｒｉｌｏ
ｎ （ＡＴＣＣ２４７２５）の分生胞子を、均一に接種
することができ、接種した分生胞子は、１〜２日間で発
芽し、約２週間で培養器内に蔓延した。The culture vessel was moved to 37°C and cultured. By this method, Ph
anerochaete chrysosporillo
Conidia of n (ATCC24725) could be uniformly inoculated, and the inoculated conidia germinated in 1 to 2 days and spread in the culture vessel in about 2 weeks.
実施例２
同様の方法によりＰｈａｎｅｒｏｃｈａｅｔｅ ｃｈｒ
ｙｓｏｓｐｏｒｉｕｍ（ＡＴＣＣ３４５４１）の分生胞
子を均一に接種することができ、接種した分生胞子は、
１〜２日間で発芽し、約２週間で培養器内に蔓延した。Example 2 Phanerochaete chr by similar method
ysosporium (ATCC34541) can be uniformly inoculated, and the inoculated conidia are
The seeds germinated in 1 to 2 days and spread within the culture vessel in about 2 weeks.
実施例３
実施例１において、無菌生理食塩水に胞子を懸濁するか
わりに栄養源を含む溶液（溶液Ａ）に胞子を懸濁させ、
菌を接種した場合、胞子は１〜２日間で発芽し、３〜４
日間で培養器内に蔓延した。Example 3 In Example 1, instead of suspending the spores in sterile physiological saline, the spores were suspended in a solution containing a nutrient source (solution A),
When inoculated with bacteria, spores germinate in 1 to 2 days, and spores germinate in 3 to 4 days.
It spread within the incubator within days.
？容液Ａ
グルコース ５．０ｇ／ （！ＮＨ，ＩＩ□
ＰＯ４２，Ｏｇ／ １Ｉ
ＣｕＳＯｓ４Ｈｚ０ １ ｍｇ／ ｅＭｎＳ
Ｏｌ・８１１２ｏ ５ ｍｇ／ ＩＦｅ（
ＣＪＩｓＯｔ）３１１ｚ０ １２ ｍｇ／ ７ＩＣ
ａＣｅ ｚ ・２１１ｚＯ７３，６ｍｇ／ ｌＭｇＳＯ
４・７１１２０ ０．５８／ ｆｆＫｚＨＰ
Ｏ４０，４ｇ／ １１
ＫＩＩＺＰＯ４０，６ｇ／ １
酵母エキス ０．１ｇ／ １塩酸チアミン
１００μｇ／ｆｆＣｏＣｊ！ ｚ・６Ｈｚ０
１ ｍｇ／ ｌＺｎ５Ｏｎ４Ｈｚ０
６．６ ｍｇ／ １ｐＨ５，５
実施例４
実施例２において、無菌生理食塩水に胞子を懸濁するか
わり実施例３記載の栄養源を含む溶液Ａに胞子を懸濁さ
せ菌を接種した場合、胞子は１〜２日間で発芽し、３〜
４日間で培養器内に蔓延した。? Liquid A Glucose 5.0g/ (!NH,II□
PO42, Og/ 1I CuSOs4Hz0 1 mg/eMnS
Ol・8112o 5 mg/IFe(
CJIsOt) 311z0 12 mg/7IC
aCe z ・211zO73,6mg/lMgSO
4・71120 0.58/ ffKzHP
O40.4g/11 KIIZPO40.6g/1 Yeast extract 0.1g/1 Thiamine hydrochloride
100μg/ffCoCj! z・6Hz0
1 mg/lZn5On4Hz0
6.6 mg/1 pH 5,5 Example 4 In Example 2, when the spores were suspended in solution A containing the nutrient source described in Example 3 instead of being suspended in sterile physiological saline and the bacteria were inoculated, Spores germinate in 1-2 days, and 3-
The virus spread within the culture vessel within 4 days.
リグノセルロース物質に高温下で胞子懸濁液を送入する
接種法を用いることにより次の効果がある。The use of an inoculation method in which a spore suspension is introduced into lignocellulosic material at high temperatures has the following effects.
１、他の微生物の混入を防ぐことができる。1. Contamination with other microorganisms can be prevented.
２、リグノセルロース物質の冷却工程を省くことができ
、パルプ化コストの低減が行える。2. The cooling process of the lignocellulose material can be omitted, reducing pulping costs.
３、均一に微生物を接種することにより、短期間に菌糸
が蔓延し、リグノセルロース物質中の脱リグニンを早め
る。すなわち、パルプ化工程を短期間にすることができ
る。3. By uniformly inoculating microorganisms, mycelium spreads in a short period of time, accelerating delignification in lignocellulosic materials. That is, the pulping process can be shortened.
４、胞子の懸濁液に栄養源を含むものを用いた場合、紙
バルブ産業のスチーミング時にパルプ化薬液を送入した
際と同じ原理で栄養源がリグノセルロース物質の内部ま
でしみわたり、微生物の繁殖をよくする。4. When a spore suspension containing a nutrient source is used, the nutrient source permeates into the interior of the lignocellulosic material using the same principle as when pulping chemicals are introduced during steaming in the paper valve industry, and the microorganisms are Improve breeding.
以上の効果によりバイオパルピングを効率化することが
できる。Due to the above effects, biopulping can be made more efficient.
第１図は、本願発明を実施するための装置の略図である
。FIG. 1 is a schematic diagram of an apparatus for carrying out the present invention.
Claims (1)
ンを分解する方法において、リグノセルロース物質を加
熱滅菌し、次いで該リグノセルロース物質に微生物の懸
濁液を高温下で接触せしめることを特徴とする微生物の
リグノセルロース物質への接種方法。 ２、微生物が分生胞子を形成する担子菌であるファネロ
ケーテ・クリソスポリウム（Ｐｈａｎｅｒｏｃｈａｅ−
ｔｅ ｃｈｒｙｓｏｓｐｏｒｉｕｍ）であることを特徴
とする特許請求の範囲第１項記載の接種方法。 ３、微生物の懸濁液が胞子を含む液であることを特徴と
する特許請求の範囲第１項記載の接種方法。 ４、微生物の懸濁液が栄養源を含む液であることを特徴
とする特許請求の範囲第１項記載の接種方法。 ５、リグノセルロース物質がチップ状、あるいは繊維状
であることを特徴とする特許請求の範囲第１項記載の方
法。[Claims] 1. A method for decomposing lignin by allowing microorganisms to act on a lignocellulosic material, which involves heat sterilizing the lignocellulosic material, and then contacting the lignocellulosic material with a suspension of microorganisms at a high temperature. A method for inoculating lignocellulosic material with microorganisms characterized by: 2. Phanerochae chrysosporium, a basidiomycete in which the microorganism forms conidia
te chrysosporium). 3. The inoculation method according to claim 1, wherein the suspension of microorganisms is a liquid containing spores. 4. The inoculation method according to claim 1, wherein the suspension of microorganisms is a liquid containing a nutrient source. 5. The method according to claim 1, wherein the lignocellulosic material is in the form of chips or fibers.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61237223A JP2568522B2 (en) | 1986-10-07 | 1986-10-07 | Method of inoculating microorganisms to lignocellulosic material |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61237223A JP2568522B2 (en) | 1986-10-07 | 1986-10-07 | Method of inoculating microorganisms to lignocellulosic material |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS6391077A true JPS6391077A (en) | 1988-04-21 |
| JP2568522B2 JP2568522B2 (en) | 1997-01-08 |
Family
ID=17012203
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP61237223A Expired - Fee Related JP2568522B2 (en) | 1986-10-07 | 1986-10-07 | Method of inoculating microorganisms to lignocellulosic material |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2568522B2 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1991010773A1 (en) * | 1990-01-19 | 1991-07-25 | Kabushiki Kaisha Kobe Seiko Sho | Process for producing pulp |
| JP2004269351A (en) * | 2003-03-06 | 2004-09-30 | Yuen Foong Yu Paper Mfg Co Ltd | Nutrient liquid for plant and its preparing method |
| CN106811422A (en) * | 2017-02-15 | 2017-06-09 | 安吉国千环境科技有限公司 | Microbial inoculum cultural method and the technique for carrying out stalk fermentation pretreatment using the microbial inoculum |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108384723B (en) * | 2018-03-13 | 2020-11-10 | 中国林业科学研究院亚热带林业研究所 | Preparation method and application of compound microbial agent |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5379085A (en) * | 1976-10-20 | 1978-07-13 | Gen Electric | Removing of lignin by biological pretreatment of lignocellulose |
-
1986
- 1986-10-07 JP JP61237223A patent/JP2568522B2/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5379085A (en) * | 1976-10-20 | 1978-07-13 | Gen Electric | Removing of lignin by biological pretreatment of lignocellulose |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1991010773A1 (en) * | 1990-01-19 | 1991-07-25 | Kabushiki Kaisha Kobe Seiko Sho | Process for producing pulp |
| JP2004269351A (en) * | 2003-03-06 | 2004-09-30 | Yuen Foong Yu Paper Mfg Co Ltd | Nutrient liquid for plant and its preparing method |
| CN106811422A (en) * | 2017-02-15 | 2017-06-09 | 安吉国千环境科技有限公司 | Microbial inoculum cultural method and the technique for carrying out stalk fermentation pretreatment using the microbial inoculum |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2568522B2 (en) | 1997-01-08 |
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