JP4204217B2 - Cellulase production substrate - Google Patents
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- JP4204217B2 JP4204217B2 JP2001337383A JP2001337383A JP4204217B2 JP 4204217 B2 JP4204217 B2 JP 4204217B2 JP 2001337383 A JP2001337383 A JP 2001337383A JP 2001337383 A JP2001337383 A JP 2001337383A JP 4204217 B2 JP4204217 B2 JP 4204217B2
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Description
【０００１】
【発明の属する技術分野】
本発明は、セルロース生産用基質、その製造方法、該基質を用いたセルラーゼの製造方法、およびセルロースの分解または糖化方法に関する。
【０００２】
【従来技術】
セルラーゼは、セルロースを、グルコース、セロビオースやセロオリゴトースに加水分解する酵素反応系を触媒する酵素群の総称であり、その作用様式により、Ｃ１酵素（アビセラーゼ、セロビオヒドラーゼ、ＦＰアーゼ、エキソ−β−グルカナーゼ等とも呼称されている。）、Ｃｘ酵素（ＣＭＣアーゼ、エンド−β−グルカナーゼともいう。）およびβ−グルコシダーゼ（セロビアーゼともいう。）など種々の名称で呼ばれる酵素が存在する。セルラーゼはこれら酵素の相互作用により、セルロースを最終的にはグルコースまで分解する。
【０００３】
一方、近年、セルラーゼを用いてセルロース系バイオマス資源を酵素分解、糖化することにより構成単位であるグルコース、キシロースにし、更にこれを発酵して得られるエタノールや乳酸などを液体燃料もしくは化学原料として利用することが検討されている。
しかし、糖化に用いるセルラーゼの生産に用いる炭素源としては、安価なセルロース資源を用いることが好ましいが、安価に入手できるセルロース資源をそのまま用いたのでは十分な生産性が得られないという欠点があった。このため、従来より粉末セルロース等の純セルロースが基質として用いられているが、この方法はセルラーゼ生産性は高いものの、高価格であり、セルラーゼ生産コストの大部分を占めてしまうという難点があった。
安価なバイオマスの基質用の処理方法としては、例えば、バガスを苛性アルカリで蒸煮した後、次亜塩素酸系化合物で処理するセルラーゼ生産用基質の製造法（特公平５−３３９８４号公報）が報告されているが、国内で大量に排出され、入手が容易な古紙を、セルラーゼ生産用基質として有効に用いることができる技術は未だ見いだされていない。
【０００４】
【発明が解決しようとする課題】
本発明の課題は、上記従来技術の問題点を解消しようとするものであって、国内で大量に排出され、入手が容易な古紙を、セルラーゼ生産用基質として有効に用いる手段を提供し、これにより、セルラーゼの製造、およびセルロースの分解、糖化を、安価にかつ効率的に行うことを目的とするものである。
【０００５】
【課題を解決するための手段】
本発明者は、上記課題を解決するため鋭意研究の結果、古紙を硫酸第一鉄で蒸煮処理し、これを炭素源としてセルラーゼ生産菌を培養すると、セルラーゼの生産性が著しく向上し、セルラーゼを安価かつ効率的に製造できるとともに、セルロースの分解、糖化も、安価、効率的に行いうることを見いだし、本発明を完成させたものである。
【０００６】
すなわち、本発明は以下（１）〜（８）に係るものである。
（１）古紙を硫酸第一鉄溶液中で蒸煮処理することを特徴とするセルラーゼ生産用基質の製造方法。
（２）古紙が乾燥重量基準で有機物比率９０％以上、リグニン含有量２０％以下、ヘミセルロース含有量２０％以下の組成を有するものであることを特徴とする（１）記載のセルラーゼ生産用基質の製造方法。
（３）古紙がオフィス古紙または段ボール古紙であることを特徴とする（１）記載のセルラーゼ生産用基質の製造方法。
（４）（１）〜（３）のいずれか一項記載の製造方法により得られるセルラーゼ生産用基質。
（５）（４）に記載のセルラーゼ生産用基質を炭素源として、セルラーゼ生産菌を培養することによりセルラーゼを産生せしめることを特徴とするセルラーゼの製造方法。
（６）（５）に記載の方法により得られたセルラーゼを用いてセルロースを分解または糖化することを特徴とするセルロースの分解または糖化方法。
（７）セルラーゼ生産菌がアクレモニウム属に属する微生物である（５）に記載の方法。
（８）アクレモニウム属に属する微生物がアクレモニウム・セルロリティカス（Acremonium cellulolyticus）Ｃ１株（ＦＥＲＭ Ｐ−１８５０８）である（７）に記載の方法。
【０００７】
本発明において用いる古紙は特にその種類を問わないが、例えばオフィス等で使用されたコピー用紙等のオフィス古紙、段ボール古紙、古新聞紙等が挙げられる。
古紙を用いてセルラーゼ生産用基質を製造する場合、古紙をシュレッデイングあるいは裁断し、硫酸第１鉄溶液中で蒸煮処理する。この際、他の薬品処理、あるいは叩解（リフアイニング）法、凍結粉砕、ボールミルやロールミル等による機械的な破砕処理等の前処理は特に必要としないが、本発明において使用する古紙類としては、乾燥重量基準で、有機物比率が９０％以上、リグニン含有量が２０％以下、ヘミセルロース含有量が２０％以下である場合に、特に効果的である．すなわち、リグニン含有量が２０％以下、ヘミセルロース含有量が２０％以下であると、セルロース成分が多くなるため、より効率的にセルラーゼ生産用基質を得ることができる。
【０００８】
本発明で使用される古紙は、シュレツデイングもしくは裁断によって３０ｍｍ×１００ｍｍ以下にして使用するのが好ましい。また、硫酸第１鉄の使用量は、古紙に対して０．０１％〜１重量％、好ましくは０．０５〜１％である。具体的には、硫酸第１鉄溶液として、ＦｅＳＯ４・７Ｈ２Ｏの０．００１〜０．５％水溶液を用い、古紙は当該水溶液中に２〜２０％、好ましくは５〜１５％加えられる。また、蒸煮温度は８０〜１３０℃が好ましい。
【０００９】
本発明において、セルラーゼ生産用基質を製造するには、例えば硫酸第一鉄水溶液の中に上記シュレッデイングあるいは裁断した古紙を加え、十分に浸漬し、８０℃〜１３０℃、より好ましくは１００℃〜１２５℃で１分以上、好ましく５〜１５分蒸煮処理を行ったのち、ケーキを充分、水洗し、セルラーゼ生産用基質を得る。
本発明のセルラーゼ生産用基質は、セルラーゼ生産菌の炭素源として極めて良好なものであり、本発明のセルラーゼ生産用基質を用いることにより、セルラーゼ生産菌は、セルラーゼを培地中に大量に生産するとともに、産生されるセルラーゼは、ＦＰアーゼ、ＣＭＣアーゼおよびセロビアーゼのすべてにおいて活性が高いので、本発明のセルラーゼ生産用基質を用いてセルラーゼ生産菌を培養することにより、セルラーゼを効率的に生産することが可能となり、得られたセルラーゼは、極めて効果的にセルロース原料を糖化、分解できる。また、本発明においては、上記セルラーゼ生産用基質、あるいはこれを主炭素として他のセルロース原料を培地に加え、該培地にセルラーゼ生産菌を培養することにより、直接、セルロースを分解、糖化することもできる。他のセルロース原料としては、バガス、イネワラ、もみがらあるいは無処理の古紙等が挙げられる。
【００１０】
本発明において、これらセルラーゼの製造あるいはセルロースの分解、糖化に使用するセルラーゼ生産菌は特に限定されず、従来、セルラーゼ生産菌として知られていた、トリコデルマ属、アスペルギルス属、スポロトリクム属、アクレモニウム属に属する微生物等が用いられるが、その中でもアクレモニウム属に属する微生物が好ましく、特に好ましいものはアクレモニウム・セルロティカス（Acremonium cellulolyticus）Ｃ１株（ＦＥＲＭ Ｐ−１８５０８）である。
【００１１】
【実施例】
以下、本発明の実施例を示すが、本発明は特にこれにより限定されるものではない。
実施例１
＜セルラーゼ生産用基質の調整＞
シュレツダー（panasonic KX−221C）で３mm×５Omm程度に裁断した使用済オフィス紙５０gを、０．０１％の硫酸第一鉄７水和物水溶液５００ｍＬ中に添加し、ガラス棒で浸漬させた後、オートクレーブを用いて１２１℃、１０分間加熱処理した。次いでこの加熱処理した古紙スラリーを１０００ｍＬのイオン交換水で水洗し、吸引濾過した。次いで生成したケーキを８０℃で２４時間乾燥し、硫酸第一鉄で処理した古紙を得た。
次に、比較用セルラーゼ生産用基質として、無処理オフィス紙、加熱処理オフィス紙、硫酸処理オフィス紙を以下のように調製した。
【００１２】
＜比較用セルラーゼ生産用基質の調製＞
（１）無処理オフィス紙；シュレツデイングしたオフィス紙をそのまま炭素源として使用した。
（２）加熱処理オフィス紙；シュレツデイングしたオフィス紙５０gに５００ｍＬイオン交換水を添加し、ガラス棒で浸漬させた後、１２１℃、１０分間加熱処理した後、１０００ｍＬのイオン交換水で水洗し、吸引濾過して得られたケーキを８０℃で２４時間乾燥して調製した。
（３）硫酸処理オフィス紙；シュレツデイングしたオフィス紙５０gに５０OｍＬの０．０１％硫酸を添加し、ガラス棒で浸溝させた後、１２１℃、１０分間加熱処理した後、１０００ｍＬのイオン交換水で水洗し、吸引濾過して得られたケーキを８０℃で２４時間乾燥して調製した。
これらとは別に、比較用セルラーゼ生産用基質として（４）粉末セルロース（Solka Floc BW200）を用意した。
【００１３】
本発明における硫酸第１鉄使用のセルラーゼ生産用基質、および上記（１）〜（４）の比較用セルラーゼ生産用基質のそれぞれを炭素源として、以下の組成を有するセルラーゼ生産菌培養培地を調整した。
〔培地の組成〕：
炭素源（セルラーゼ生産用基質） ５０ ｇ／Ｌ
硫酸アンモニウム ５ ｇ／Ｌ
コーンステイーブリカー １０ ｇ／Ｌ
尿素 ３ ｇ／Ｌ
硫酸マグネシウム １．２ｇ／Ｌ
リン酸２水素カリウム １２ ｇ／Ｌ
硫酸亜鉛 １０ ｇ／Ｌ
硫酸マンガン １０ ｇ／Ｌ
硫酸銅 １０ ｇ／Ｌ
Ｔｗｅｅｎ８０ １ ｇ／Ｌ
ｐＨ ４．０
【００１４】
＜セルラーゼの生産培養試験＞
上記組成の各液体培地にセルラーゼ生産菌であるアクレモニウム・セルロリティカス（Acremonium cellulolyticus）Ｃ１株（ＦＥＲＭ Ｐ−１８５０８）を接種して３０℃で５日間培養した。この培養液を遠心分離して得た上澄液について、生産されたセルラーゼにおけるFPアーゼ、CMCアーゼ及びセロビアーゼの活性を以下の方法に基づき測定した。
【００１５】
＜セルラーゼ活性測定法＞
FPアーゼ：濾紙（ワットマンNo．1）５０mgを基質とし、これに酵素液０．５mLとクエン酸緩衝液（ｐＨ４．８、０．０５M）１．０mLを加え、５０℃で１．０時間酵素反応を行った後、ジニトロサリチル酸試薬３．０mLを加え、１００℃で５分間加熱し発色させる。冷却後、これにイオン交換水または蒸留水２０mLを加え５４０ｎmの波長で比色定量する。１分間に１μmolのグルコースに相当する還元糖を生成する酵素量を１ユニットとした。
CMCアーゼ：クエン酸緩衝液（ｐＨ４．８、０．０５M）に溶解した２％カルボキシメチルセルロースナトリウム塩溶液に対し、等量の適当に希釈した酵素溶液を加え、５０℃で３０分間酵素反応を行った後、ジニトロサリチル酸試薬３．０mLを加え１００℃で５分間加熱し酵素反応を停止させ、これにイオン交換水または蒸留水２０mLを加え５４０ｎｍの波長で比色定量する。１分間に１μmolのグルコースに相当する還元糖を生成する酵素量を１ユニットとした。
セロビアーゼ活性：クエン酸緩衝液（ｐＨ４．８、０．０５M）に１５mM溶解したセロビオース溶液１ｍＬに対し、等量の適当に希釈した酵素溶液を加え、５０℃で３０分間酵素反応を行った後、１００℃で５分間加熱し酵素反応を停止させ、液中のグルコース量をグルコース測定キット（和光純薬製グルコスタットB）で測定した。１分間に１μmolのグルコースに相当する還元糖を生成する酵素量を１ユニットとした。
得られた結果を表１に示す。
【００１６】
【表１】
【００１７】
【発明の効果】
本発明におけるセルラーゼ生産用基質は、古紙を原料としていながら、純セルロースを使用する場合に匹敵するセルラーゼ生産効率を示す。しかも、その製造法は、古紙を裁断後、単に硫酸鉄溶液中で蒸煮するのみであって、極めて簡便なものであり、本発明はセルラーゼ生産およびセルロース原料の分解、糖化において、極めて安価で実用的な手段を提供しうる点で画期的なものである。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a substrate for producing cellulose, a method for producing the same, a method for producing cellulase using the substrate, and a method for decomposing or saccharifying cellulose.
[0002]
[Prior art]
Cellulase, cellulose, glucose, is a generic name for enzymes that catalyze the hydrolyzing enzyme reaction system cellobiose and Seroorigotosu by its mode of action, C 1 enzyme (avicelase, cellobiohydrolase, FP ase, exo - There are enzymes called various names such as β-glucanase etc.), Cx enzyme (CMCase, also called endo-β-glucanase) and β-glucosidase (also called cellobiase). Cellulase eventually breaks cellulose down to glucose by the interaction of these enzymes.
[0003]
On the other hand, in recent years, cellulosic biomass resources are enzymatically decomposed and saccharified to produce structural units such as glucose and xylose, which are further fermented to use ethanol, lactic acid, etc. as liquid fuels or chemical raw materials. It is being considered.
However, it is preferable to use an inexpensive cellulose resource as a carbon source used to produce cellulase used for saccharification, but there is a disadvantage that sufficient productivity cannot be obtained if a cellulose resource available at low cost is used as it is. It was. For this reason, pure cellulose such as powdered cellulose has been conventionally used as a substrate. However, this method has high cellulase productivity, but is expensive and occupies most of the cellulase production cost. .
As a processing method for an inexpensive biomass substrate, for example, a method for producing a substrate for cellulase production in which bagasse is steamed with caustic and then treated with a hypochlorous acid compound (Japanese Patent Publication No. 5-33984) is reported. However, no technology has yet been found that can effectively use used paper, which is discharged in large quantities in Japan and easily available, as a substrate for cellulase production.
[0004]
[Problems to be solved by the invention]
An object of the present invention is to solve the above-mentioned problems of the prior art, and provides a means for effectively using waste paper that is discharged in a large amount and easily available as a substrate for cellulase production. Thus, it is intended to perform cellulase production and cellulose decomposition and saccharification at low cost and efficiently.
[0005]
[Means for Solving the Problems]
As a result of diligent research to solve the above problems, the present inventor, when used paper was steamed with ferrous sulfate and cellulase-producing bacteria were cultured using this as a carbon source, the productivity of cellulase was remarkably improved. The present invention has been completed by finding that it can be produced inexpensively and efficiently, and that cellulose can be decomposed and saccharified inexpensively and efficiently.
[0006]
That is, the present invention relates to the following (1) to (8).
(1) A method for producing a substrate for cellulase production, wherein used paper is steamed in a ferrous sulfate solution.
(2) The cellulase production substrate according to (1), wherein the waste paper has a composition having an organic matter ratio of 90% or more, a lignin content of 20% or less, and a hemicellulose content of 20% or less on a dry weight basis. Production method.
(3) The method for producing a substrate for cellulase production according to (1), wherein the waste paper is office waste paper or cardboard waste paper.
(4) A cellulase production substrate obtained by the production method according to any one of (1) to (3).
(5) A method for producing cellulase, wherein cellulase is produced by culturing cellulase-producing bacteria using the cellulase-producing substrate according to (4) as a carbon source.
(6) A method for decomposing or saccharifying cellulose, comprising decomposing or saccharifying cellulose using the cellulase obtained by the method according to (5).
(7) The method according to (5), wherein the cellulase-producing bacterium is a microorganism belonging to the genus Acremonium.
(8) The method according to (7) , wherein the microorganism belonging to the genus Acremonium is Acremonium cellulolyticus C1 strain (FERM P-18508).
[0007]
The waste paper used in the present invention is not particularly limited, and examples thereof include office waste paper such as copy paper used in offices, corrugated waste paper, old newspaper and the like.
When manufacturing a substrate for cellulase production using waste paper, the waste paper is shredded or cut and steamed in a ferrous sulfate solution. At this time, pretreatment such as other chemical treatment or beating (refining) method, freeze pulverization, mechanical crushing treatment by a ball mill, a roll mill or the like is not particularly required, but the used paper used in the present invention is dried. This is particularly effective when the organic matter ratio is 90% or more, the lignin content is 20% or less, and the hemicellulose content is 20% or less on a weight basis. That is, when the lignin content is 20% or less and the hemicellulose content is 20% or less, the cellulose component increases, so that a cellulase production substrate can be obtained more efficiently.
[0008]
The used paper used in the present invention is preferably 30 mm × 100 mm or less by shredding or cutting. Moreover, the usage-amount of ferrous sulfate is 0.01 to 1 weight% with respect to a used paper, Preferably it is 0.05 to 1%. Specifically, a 0.001 to 0.5% aqueous solution of FeSO 4 · 7H 2 O is used as the ferrous sulfate solution, and waste paper is added to the aqueous solution in an amount of 2 to 20%, preferably 5 to 15%. . The cooking temperature is preferably 80 to 130 ° C.
[0009]
In the present invention, in order to produce a substrate for cellulase production, for example, the above-mentioned shredded or cut waste paper is added to an aqueous ferrous sulfate solution, and sufficiently immersed, and 80 ° C to 130 ° C, more preferably 100 ° C to 130 ° C. After performing the steaming treatment at 125 ° C. for 1 minute or more, preferably 5 to 15 minutes, the cake is sufficiently washed with water to obtain a substrate for cellulase production.
The cellulase-producing substrate of the present invention is extremely good as a carbon source for cellulase-producing bacteria. By using the cellulase-producing substrate of the present invention, the cellulase-producing bacteria can produce cellulase in a large amount in a medium. Since the cellulase produced is highly active in all of FPase, CMCase and cellobiase, cellulase can be efficiently produced by culturing cellulase-producing bacteria using the cellulase-producing substrate of the present invention. The resulting cellulase can saccharify and decompose the cellulose raw material very effectively. Further, in the present invention, cellulose may be directly decomposed and saccharified by adding the above-mentioned cellulase production substrate or another cellulose raw material to the medium as a main carbon and culturing cellulase-producing bacteria in the medium. it can. Examples of other cellulose raw materials include bagasse, rice bran, rice husk or untreated waste paper.
[0010]
In the present invention, the cellulase-producing bacteria used for the production of these cellulases or the degradation or saccharification of cellulose are not particularly limited, and are conventionally known as cellulase-producing bacteria, such as Trichoderma, Aspergillus, Sporotricum, and Acremonium. Among them, microorganisms belonging to the genus Acremonium are preferable, and Acremonium cellulolyticus C1 strain (FERM P-18508) is particularly preferable.
[0011]
【Example】
Examples of the present invention will be described below, but the present invention is not particularly limited thereby.
Example 1
<Preparation of substrate for cellulase production>
After adding 50 g of used office paper cut to about 3 mm × 5 Omm with a Schretzder (panasonic KX-221C) into 500 mL of 0.01% ferrous sulfate heptahydrate aqueous solution, It heat-processed for 121 minutes at 121 degreeC using the autoclave. Next, the heat-treated waste paper slurry was washed with 1000 mL of ion exchange water and suction filtered. Next, the produced cake was dried at 80 ° C. for 24 hours to obtain waste paper treated with ferrous sulfate.
Next, untreated office paper, heat-treated office paper, and sulfuric acid-treated office paper were prepared as follows as a substrate for producing cellulase for comparison.
[0012]
<Preparation of cellulase production substrate for comparison>
(1) Untreated office paper; shredded office paper was used as it was as a carbon source.
(2) Heat-treated office paper: After adding 500 mL ion-exchanged water to 50 g of shredded office paper and immersing it in a glass rod, heat-treated at 121 ° C. for 10 minutes, and then washed with 1000 mL of ion-exchanged water. The cake obtained by suction filtration was prepared by drying at 80 ° C. for 24 hours.
(3) Sulfuric acid-treated office paper; 50OmL of 0.01% sulfuric acid was added to 50g of shredded office paper, immersed in a glass rod, heat-treated at 121 ° C for 10 minutes, and then 1000mL of ion exchange A cake obtained by washing with water and suction filtration was prepared by drying at 80 ° C. for 24 hours.
Separately from these, (4) powdered cellulose (Solka Floc BW200) was prepared as a cellulase production substrate for comparison.
[0013]
A cellulase-producing bacterial culture medium having the following composition was prepared using each of the cellulase-producing substrate using ferrous sulfate and the comparative cellulase-producing substrates (1) to (4) in the present invention as a carbon source. .
[Composition of medium]:
Carbon source (substrate for cellulase production) 50 g / L
Ammonium sulfate 5 g / L
Cornstay Briker 10 g / L
Urea 3 g / L
Magnesium sulfate 1.2g / L
Potassium dihydrogen phosphate 12 g / L
Zinc sulfate 10 g / L
Manganese sulfate 10 g / L
Copper sulfate 10 g / L
Tween80 1 g / L
pH 4.0
[0014]
<Cellulase production culture test>
Each liquid medium having the above composition was inoculated with cellulase-producing bacteria Acremonium cellulolyticus C1 strain (FERM P-18508) and cultured at 30 ° C. for 5 days. For the supernatant obtained by centrifuging this culture solution, the activities of FPase, CMCase and cellobiase in the produced cellulase were measured based on the following method.
[0015]
<Cellulase activity measurement method>
FPase: 50 mg of filter paper (Whatman No. 1) as a substrate, 0.5 mL of enzyme solution and 1.0 mL of citrate buffer (pH 4.8, 0.05 M) were added thereto, and the enzyme was added at 50 ° C. for 1.0 hour. After the reaction, 3.0 mL of a dinitrosalicylic acid reagent is added, and the color is developed by heating at 100 ° C. for 5 minutes. After cooling, 20 mL of ion-exchanged water or distilled water is added thereto, and colorimetric determination is performed at a wavelength of 540 nm. The amount of enzyme that produces reducing sugar corresponding to 1 μmol of glucose per minute was defined as 1 unit.
CMCase: To 2% carboxymethylcellulose sodium salt solution dissolved in citrate buffer (pH 4.8, 0.05M), add an equal amount of an appropriately diluted enzyme solution and carry out the enzyme reaction at 50 ° C for 30 minutes. Thereafter, 3.0 mL of a dinitrosalicylic acid reagent is added and heated at 100 ° C. for 5 minutes to stop the enzyme reaction, and 20 mL of ion-exchanged water or distilled water is added thereto and colorimetrically determined at a wavelength of 540 nm. The amount of enzyme that produces reducing sugar corresponding to 1 μmol of glucose per minute was defined as 1 unit.
Cellobiase activity: To 1 mL of cellobiose solution dissolved in 15 mM of citrate buffer (pH 4.8, 0.05 M), an equal amount of an appropriately diluted enzyme solution was added, and an enzyme reaction was performed at 50 ° C. for 30 minutes. The enzyme reaction was stopped by heating at 100 ° C. for 5 minutes, and the amount of glucose in the liquid was measured with a glucose measurement kit (Glucostat B manufactured by Wako Pure Chemical Industries). The amount of enzyme that produces reducing sugar corresponding to 1 μmol of glucose per minute was defined as 1 unit.
The obtained results are shown in Table 1.
[0016]
[Table 1]
[0017]
【The invention's effect】
The substrate for cellulase production in the present invention exhibits cellulase production efficiency comparable to that when pure cellulose is used while using waste paper as a raw material. Moreover, the production method is simply simple, after cutting waste paper, and then steamed in an iron sulfate solution, and the present invention is extremely inexpensive and practical in cellulase production and decomposition and saccharification of cellulose raw materials. It is groundbreaking in that it can provide a practical means.
Claims (8)
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| JP2011041522A (en) | 2009-08-21 | 2011-03-03 | Asahi Breweries Ltd | METHOD FOR PRODUCING beta-GLUCANASE AND XYLANASE WITH WHEAT BRAN AND LIQUID CULTURE MEDIUM |
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