JP2014147304A - Enzyme action-promoting agent to promote polysaccharide-degrading enzyme activity and organic waste-decomposing agent containing the same - Google Patents
Enzyme action-promoting agent to promote polysaccharide-degrading enzyme activity and organic waste-decomposing agent containing the same Download PDFInfo
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Abstract
Description
本発明は、多糖類分解酵素の活性促進剤及びそれを含有する有機残渣分解剤に関する。 The present invention relates to an activity promoting agent for polysaccharide-degrading enzymes and an organic residue decomposing agent containing the same.
一般に、植物体を構成する多糖類、例えばセルロース及びキシランを加水分解する酵素として多糖類分解酵素、例えばセルラーゼ及びキシラナーゼが知られている。これらの酵素は、例えば圃場及び各事業所等から排出される収穫残渣、家畜排泄物、及び下水余剰汚泥、並びに一般家庭から排出される食品廃棄物等の有機残渣の減量化又は堆肥化等の再処理のために利用することができる。また、将来バイオエタノールの生産に関し、上記植物性廃棄物、木材及び草材等の植物由来のバイオマスから糖を得る方法に利用されることが期待されている。また、衣料用洗浄成分として配合されたり、衣類の材質改善にも利用されている。また、植物細胞をプロトプラスト化するための研究用試薬としても使用されている。 In general, polysaccharide-degrading enzymes such as cellulases and xylanases are known as enzymes that hydrolyze polysaccharides such as cellulose and xylan that constitute plant bodies. These enzymes, for example, reduce organic waste such as harvest residues, livestock excrement, and sewage surplus sludge discharged from farms and offices, and food residues discharged from general households, composting, etc. Can be used for reprocessing. Further, in the future, bioethanol production is expected to be used in a method for obtaining sugar from plant-derived biomass such as plant waste, wood, and grass. Moreover, it is mix | blended as a washing | cleaning component for clothes, and is utilized also for the material improvement of clothing. It is also used as a research reagent for protoplastization of plant cells.
従来より、特許文献１，２に記載される多糖類分解酵素及びかかる多糖類分解酵素を産生する微生物が知られている。例えば、有機残渣を分解するに際し、微生物を利用する場合、微生物がその場所に定着、増殖したのち、有機残渣を分解するための酵素を産生することにより、有機残渣が分解される。また、多糖類分解酵素を利用する場合、有機残渣に対し、酵素資材を散布及び混合することにより、所定の気温環境下において有機残渣を分解する。 Conventionally, polysaccharide degrading enzymes described in Patent Documents 1 and 2 and microorganisms producing such polysaccharide degrading enzymes are known. For example, when a microorganism is used for decomposing an organic residue, the organic residue is decomposed by producing an enzyme for decomposing the organic residue after the microorganism has settled and propagated there. Moreover, when utilizing a polysaccharide degrading enzyme, an organic residue is decomposed | disassembled in predetermined | prescribed temperature environment by spraying and mixing an enzyme material with respect to an organic residue.
例えば、有機残渣を分解するに際し、微生物を利用する場合、分解効率は周囲の環境に依存されやすく、周囲の環境が微生物の増殖に適しており、さらに増殖するための栄養分が存在する必要がある。そのため、周囲の環境において、微生物の増殖に必要な要因が一つでも欠ければ、分解効率が著しく低下する。よって、圃場によっては思うような効果が発揮されなかったり、効果にムラ等が生ずるという問題があった。一方、酵素資材においては、微生物のように生環境を整える必要がないため、微生物資材よりも扱いやすい資材といえる。しかしながら、酵素資材は、一般的に高価であり、コスト面から有機残渣に対し大量に散布することができないという問題があった。 For example, when decomposing organic residues, when microorganisms are used, the decomposition efficiency is likely to depend on the surrounding environment, the surrounding environment is suitable for the growth of microorganisms, and there is a need for nutrients to grow. . Therefore, if even one factor necessary for the growth of microorganisms is lacking in the surrounding environment, the degradation efficiency is significantly reduced. Therefore, there is a problem that the desired effect is not exhibited depending on the field, or the effect is uneven. On the other hand, enzyme materials are easier to handle than microbial materials because it is not necessary to prepare a living environment like microorganisms. However, enzyme materials are generally expensive and have a problem that they cannot be applied in large quantities to organic residues from the cost aspect.
本発明は、本発明者らの鋭意研究の結果、多糖類分解酵素にグリホサート化合物を併用することにより多糖類分解酵素の活性を促進することができることを見出したことによりなされたものである。本発明の目的は、多糖類分解酵素の活性を促進することができる多糖類分解酵素の活性促進剤及びそれを含有する有機残渣分解剤を提供することにある。 This invention is made | formed by discovering that the activity of a polysaccharide degrading enzyme can be accelerated | stimulated by using a glyphosate compound together with a polysaccharide degrading enzyme as a result of the present inventors' earnest research. An object of the present invention is to provide a polysaccharide-degrading enzyme activity promoter capable of promoting the activity of a polysaccharide-degrading enzyme and an organic residue decomposing agent containing the same.
上記目的を達成するために本発明の一態様である多糖類分解酵素の活性促進剤は、グリホサート化合物を含有することを特徴とする。前記グリホサート化合物は、グリホサートイソプロピルアミン塩、グリホサートアンモニウム塩、グリホサートカリウム塩、及びグリホサートトリメシウム塩から選ばれる少なくとも一種であることが好ましい。 In order to achieve the above object, the polysaccharide degrading enzyme activity promoter which is one embodiment of the present invention contains a glyphosate compound. The glyphosate compound is preferably at least one selected from glyphosate isopropylamine salt, glyphosate ammonium salt, glyphosate potassium salt, and glyphosate trimesium salt.
本発明の別態様である有機残渣分解剤は、グリホサート化合物を含有する多糖類分解酵素の活性促進剤、及び多糖類分解酵素を含有する。前記多糖類分解酵素は、セルラーゼ及びキシラナーゼから選ばれる少なくとも一種であることが好ましい。 The organic residue decomposing agent which is another embodiment of the present invention contains an activity promoter for polysaccharide degrading enzymes containing a glyphosate compound, and a polysaccharide degrading enzyme. The polysaccharide degrading enzyme is preferably at least one selected from cellulase and xylanase.
本発明によれば、多糖類分解酵素の活性を促進することができる。 According to the present invention, the activity of a polysaccharide-degrading enzyme can be promoted.
（第１の実施形態）
以下、本発明の多糖類分解酵素の活性促進剤を具体化した第１の実施形態を詳細に説明する。
(First embodiment)
Hereinafter, a first embodiment of the polysaccharide degrading enzyme activity promoter of the present invention will be described in detail.
本実施形態のグリホサート化合物を含有する多糖類分解酵素の活性促進剤は、多糖類分解酵素と併用することにより多糖類分解酵素の活性を促進する作用を発揮する。グリホサート化合物は、Ｎ−ホスホノメチルグリシンとも呼ばれ、グリシンにリン酸を導入した誘導体である。グリホサート化合物として、例えば、グリホサートイソプロピルアミン塩、グリホサートアンモニウム塩、グリホサートカリウム塩、及びグリホサートトリメシウム塩が挙げられる。 The polysaccharide-degrading enzyme activity promoter containing the glyphosate compound of the present embodiment exhibits an action of promoting the activity of the polysaccharide-degrading enzyme when used in combination with the polysaccharide-degrading enzyme. The glyphosate compound is also called N-phosphonomethylglycine and is a derivative in which phosphoric acid is introduced into glycine. Examples of the glyphosate compound include glyphosate isopropylamine salt, glyphosate ammonium salt, glyphosate potassium salt, and glyphosate trimesium salt.
本実施形態の多糖類分解酵素の活性促進剤が適用される多糖類分解酵素としては、特に限定されないが、例えばデンプン又はその誘導体を加水分解するための酵素としてアミラーゼ（デンプン分解酵素）、セルロース又はその誘導体を加水分解するための酵素としてセルラーゼ（セルロース分解酵素）、ヘミセルロース又はその誘導体を加水分解するヘミセルラーゼ（ヘミセルロース分解酵素）、キシロース又はその誘導体を加水分解するためのキシラナーゼ（キシロース分解酵素）、ペクチン又はその誘導体を加水分解するペクチナーゼ（ペクチン分解酵素）、キチン又はその誘導体を加水分解するための酵素としてキチナーゼ（キチン分解酵素）等が挙げられる。これらの中で、グリホサート化合物による酵素活性化の促進作用に優れるセルラーゼ及びキシラナーゼがより好ましい。 The polysaccharide-degrading enzyme to which the polysaccharide-degrading enzyme activity promoter of the present embodiment is applied is not particularly limited. For example, amylase (starch degrading enzyme), cellulose, or the like can be used as an enzyme for hydrolyzing starch or a derivative thereof. Cellulase (cellulose-degrading enzyme) as an enzyme for hydrolyzing the derivative, hemicellulase (hemicellulose-degrading enzyme) for hydrolyzing hemicellulose or its derivative, xylanase (xylose-degrading enzyme) for hydrolyzing xylose or its derivative, Examples include pectinase (pectin-degrading enzyme) that hydrolyzes pectin or its derivative, and chitinase (chitinase) as an enzyme for hydrolyzing chitin or its derivative. Among these, cellulase and xylanase, which are excellent in the action of promoting enzyme activation by the glyphosate compound, are more preferable.
次に、上記のように構成された多糖類分解酵素の活性促進剤の作用を説明する。
本実施形態の多糖類分解酵素の活性促進剤は、有効成分であるグリホサート化合物が多糖類分解酵素の酵素活性を促進する作用を発揮する。
Next, the action of the polysaccharide degrading enzyme activity promoter configured as described above will be described.
The polysaccharide-degrading enzyme activity promoter of this embodiment exhibits an action in which the glyphosate compound as an active ingredient promotes the enzyme activity of the polysaccharide-degrading enzyme.
本実施形態の多糖類分解酵素の活性促進剤によれば、以下のような効果を得ることができる。
（１）本実施形態の多糖類分解酵素の活性促進剤は、有効成分としてグリホサート化合物を含有する。したがって、グリホサート化合物により多糖類分解酵素の活性が促進される。
According to the polysaccharide-degrading enzyme activity promoter of the present embodiment, the following effects can be obtained.
(1) The polysaccharide-degrading enzyme activity promoter of this embodiment contains a glyphosate compound as an active ingredient. Therefore, the activity of polysaccharide degrading enzyme is promoted by the glyphosate compound.
（２）本実施形態の多糖類分解酵素の活性促進剤において、グリホサート化合物は、グリホサートイソプロピルアミン塩、グリホサートアンモニウム塩、グリホサートカリウム塩、及びグリホサートトリメシウム塩が好ましい。したがって、多糖類分解酵素の活性促進作用をより向上させることができる。 (2) In the polysaccharide-degrading enzyme activity promoter of this embodiment, the glyphosate compound is preferably glyphosate isopropylamine salt, glyphosate ammonium salt, glyphosate potassium salt, and glyphosate trimesium salt. Therefore, the activity promoting action of the polysaccharide-degrading enzyme can be further improved.
なお、上記実施形態は以下のように変更してもよい。
・上記実施形態の多糖類分解酵素の活性促進剤の用途は特に限定されず、多糖類分解酵素の活性促進作用が必要な用途であればいずれも適用できる。例えば圃場及び各事業所等から排出される収穫残渣、家畜排泄物、及び下水余剰汚泥、並びに一般家庭から排出される食品廃棄物等の有機残渣の減量化又は堆肥化等の再処理のため、多糖類分解酵素と併用して利用することができる。また、将来バイオエタノールの生産に関し、上記植物性廃棄物、木材・草材等の植物由来のバイオマスから糖を得る方法に多糖類分解酵素と併用して利用することができる。また、衣料用洗浄成分及び衣類の材質改善のため、多糖類分解酵素と併用して利用することができる。また、多糖類分解酵素と併用して植物細胞をプロトプラスト化するための研究用試薬としても使用することができる。これらの中で、有機残渣の減量化のために好適に用いることができる。
In addition, you may change the said embodiment as follows.
-The use of the polysaccharide-degrading enzyme activity promoter of the above embodiment is not particularly limited, and any application that requires the activity of promoting the activity of polysaccharide-degrading enzyme is applicable. For reprocessing such as reduction or composting of organic residues such as harvest residue, livestock excrement, and sewage surplus sludge discharged from fields and business sites, etc., and food waste discharged from general households. It can be used in combination with a polysaccharide-degrading enzyme. Moreover, regarding the production of bioethanol in the future, it can be used in combination with a polysaccharide-degrading enzyme in the above-mentioned method for obtaining sugar from plant-derived biomass such as plant waste, wood and grass. In addition, it can be used in combination with a polysaccharide-degrading enzyme to improve clothing cleaning ingredients and clothing materials. It can also be used as a research reagent for protoplasting plant cells in combination with polysaccharide-degrading enzymes. In these, it can use suitably for the reduction | decrease of an organic residue.
・上記実施形態の多糖類分解酵素の活性促進剤は、本発明の効果及び使用目的を阻害しない範囲内において、水及び有機溶媒等の溶剤、並びに添加剤、例えば安定剤、保存剤、界面活性剤、及び賦形剤を適宜配合してもよい。 The polysaccharide-degrading enzyme activity promoter of the above embodiment is within the range that does not impair the effects and intended use of the present invention, and solvents such as water and organic solvents, and additives such as stabilizers, preservatives, surface activity You may mix | blend an agent and an excipient | filler suitably.
（第２の実施形態）
以下、本発明の有機残渣分解剤を具体化した第２の実施形態を詳細に説明する。
本実施形態の有機残渣分解剤は、上記第１の実施形態の多糖類分解酵素の活性促進剤及び多糖類分解酵素を含有している。多糖類分解酵素は、多糖類分解酵素の活性促進剤に含有されるグリホサート化合物と併用されることにより、有機残渣中の多糖類の分解が促進される。多糖類分解酵素としては、第１の実施形態欄に記載の酵素と同様の酵素を挙げることができる。これらの中で、グリホサート化合物による酵素活性化の促進作用に優れるセルラーゼ及びキシラナーゼがより好ましい。
(Second Embodiment)
Hereinafter, a second embodiment in which the organic residue decomposing agent of the present invention is embodied will be described in detail.
The organic residue decomposing agent of the present embodiment contains the polysaccharide decomposing enzyme activity promoter and the polysaccharide decomposing enzyme of the first embodiment. The polysaccharide-degrading enzyme is used in combination with the glyphosate compound contained in the polysaccharide-degrading enzyme activity promoter, thereby promoting the degradation of the polysaccharide in the organic residue. Examples of the polysaccharide degrading enzyme include the same enzymes as those described in the first embodiment column. Among these, cellulase and xylanase, which are excellent in the action of promoting enzyme activation by the glyphosate compound, are more preferable.
有機残渣分解剤中におけるグリホサート化合物の含有量は、多糖類分解酵素の活性促進作用向上の観点から、好ましくは０．０１〜７質量％、より好ましくは０．１〜６．５質量％、さらに好ましくは０．５〜６．３質量％である。 The content of the glyphosate compound in the organic residue decomposing agent is preferably 0.01 to 7% by mass, more preferably 0.1 to 6.5% by mass, and more preferably 0.1 to 6.5% by mass, from the viewpoint of improving the activity promoting action of the polysaccharide degrading enzyme. Preferably it is 0.5-6.3 mass%.
有機残渣分解剤中における多糖類分解酵素の含有量は、使用目的等に応じ適宜設定されるが、経済性等の観点から、好ましくは０．００１〜１０質量％、より好ましくは０．００５〜５質量％である。 The content of the polysaccharide-degrading enzyme in the organic residue decomposing agent is appropriately set according to the purpose of use and the like, but from the viewpoint of economy and the like, it is preferably 0.001 to 10% by mass, more preferably 0.005. 5% by mass.
有機残渣分解剤が適用される有機残渣としては、特に限定されず、例えば圃場及び各事業所等から排出される収穫残渣、家畜排泄物、及び下水余剰汚泥、並びに一般家庭から排出される食品廃棄物が挙げられる。 The organic residue to which the organic residue decomposing agent is applied is not particularly limited, and for example, harvest residue, livestock excrement, and excess sewage sludge discharged from fields and business establishments, and food waste discharged from general households. Things.
本実施形態の有機残渣分解剤によれば、以下のような効果を得ることができる。
（１）本実施形態の有機残渣分解剤は、グリホサート化合物及び多糖類分解酵素を含有する。したがって、有機残渣分解剤の有機残渣への投与又は散布後において、グリホサート化合物の多糖類分解酵素の活性を促進する作用により、有機残渣中の多糖類の分解が促進される。
According to the organic residue decomposing agent of this embodiment, the following effects can be obtained.
(1) The organic residue decomposing agent of this embodiment contains a glyphosate compound and a polysaccharide degrading enzyme. Therefore, after administration or application of the organic residue decomposing agent to the organic residue, the action of promoting the activity of the polysaccharide-degrading enzyme of the glyphosate compound promotes the decomposition of the polysaccharide in the organic residue.
（２）本実施形態の有機残渣分解剤は、微生物資材のように生環境を整える必要がないため、微生物資材よりも取り扱い性及び使用性を向上させることができる。
（３）特に、大量の散布が必要な圃場において、酵素の使用量を削減することができる。
(2) Since the organic residue decomposing agent of this embodiment does not need to prepare a living environment like microbial materials, it can improve handling and usability compared to microbial materials.
(3) The amount of enzyme used can be reduced, especially in fields where a large amount of spraying is required.
なお、上記実施形態は以下のように変更してもよい。
・上記実施形態の有機残渣分解剤の使用目的は特に限定されず、例えば有機残渣の減量化、及び堆肥化等の再処理のために利用することができる。
In addition, you may change the said embodiment as follows.
-The use purpose of the organic residue decomposing agent of the above embodiment is not particularly limited, and can be used for reprocessing such as reduction of organic residue and composting.
・上記実施形態の有機残渣分解剤は、本発明の効果及び使用目的を阻害しない範囲内において、水及び有機溶媒等の溶剤、並びに添加剤、例えば安定剤、保存剤、界面活性剤、及び賦形剤を適宜配合してもよい。 The organic residue decomposing agent of the above-described embodiment is within the range that does not impair the effects and intended use of the present invention, and solvents such as water and organic solvents, and additives such as stabilizers, preservatives, surfactants, and additives. You may mix | blend a dosage form suitably.
次に、実施例及び比較例を挙げて前記実施形態を更に具体的に説明する。
グリホサート化合物と多糖類分解酵素とを併用した場合におけるグリホサート化合物による多糖類分解酵素の活性促進作用を評価した。
Next, the embodiment will be described more specifically with reference to examples and comparative examples.
When the glyphosate compound and the polysaccharide degrading enzyme were used in combination, the activity promoting action of the polysaccharide degrading enzyme by the glyphosate compound was evaluated.
所定量のグリホサート化合物及び多糖類分解酵素を水に溶解することにより、表１〜４の各実施例に示される各成分濃度からなる有機残渣分解剤を調製した。尚、比較例１〜４は、多糖類分解酵素のみからなり、グリホサート化合物を配合しない構成とした。グリホサート化合物は、実施例１〜８，１７〜２４，３３〜４０，４９〜５６が日産化学工業社製のラウンドアップマックスロード（商品名）を使用した。尚、各表中においては、ラウンドアップマックスロード中のグリホサートカリウム塩濃度を示す。実施例９〜１６，２５〜３２，４１〜４８，５７〜６４においては、グリホサートイソプロピルアミン塩（ＳＩＧＭＡ社製）を使用した。多糖類分解酵素として、実施例１〜１６，４９〜６４及び比較例１，４は、セルラーゼＳＳ（ナガセケムテックス社製）を使用した。実施例１７〜３２及び比較例２は、セルラーゼＭ（メニコン社製）を使用した。実施例３３〜４８及び比較例３は、セルラーゼＡ「アマノ」３（天野エンザイム社製）を使用した。実施例１〜４８及び比較例１〜３の各有機残渣分解剤は、下記の方法により、キシラナーゼ活性を測定した。実施例４９〜６４及び比較例４の各有機残渣分解剤は、下記の方法により、カルボキシメチルセルロース分解酵素（ＣＭＣａｓｅ）活性を測定した。 By dissolving a predetermined amount of glyphosate compound and polysaccharide-degrading enzyme in water, an organic residue decomposing agent having each component concentration shown in each example of Tables 1 to 4 was prepared. In addition, Comparative Examples 1-4 consisted only of the polysaccharide degrading enzyme, and set it as the structure which does not mix | blend a glyphosate compound. As the glyphosate compound, Examples 1 to 8, 17 to 24, 33 to 40, and 49 to 56 used Roundup Max Road (trade name) manufactured by Nissan Chemical Industries, Ltd. In addition, in each table | surface, the glyphosate potassium salt density | concentration in round-up max road is shown. In Examples 9 to 16, 25 to 32, 41 to 48, and 57 to 64, glyphosate isopropylamine salt (manufactured by SIGMA) was used. Cellulase SS (manufactured by Nagase ChemteX Corporation) was used in Examples 1 to 16, 49 to 64 and Comparative Examples 1 and 4 as polysaccharide degrading enzymes. In Examples 17 to 32 and Comparative Example 2, Cellulase M (manicon) was used. In Examples 33 to 48 and Comparative Example 3, cellulase A “Amano” 3 (manufactured by Amano Enzyme) was used. Each organic residue decomposer of Examples 1-48 and Comparative Examples 1-3 measured xylanase activity by the following method. Each organic residue decomposing agent of Examples 49 to 64 and Comparative Example 4 was measured for carboxymethylcellulose degrading enzyme (CMCase) activity by the following method.
（キシラナーゼ活性測定）
ｐＨ７．０の１％キシラン（１％キシラン（ブナ材由来，ＳＩＧＭＡ社製）／５０ｍＭのＭＯＰＳ（ナカライテスク社製），１０ｍＭのＣａＣｌ２）１ｍＬに表１〜３記載の各例の試料１００μＬをそれぞれ入れ、ボルテックス後、３７℃、１５分インキュベートした。これを５００μＬずつ採り、下記組成からなるＤＮＳ溶液５００μＬにそれぞれ入れた。１００℃、５分間加熱後、放冷し、５９５ｎｍの吸光度を測定した。各種濃度のグルコース溶液を調製し、ＤＮＳ溶液で同様に反応させ、検量線を作成した。検量線から、溶出した還元糖濃度（ｍｇ／ｍＬ）を求めた。還元糖濃度は、グルコース換算し、これをキシラナーゼ活性値とした。結果を表１〜３に示す。尚、ＤＮＳ溶液は、フェノール０．２ｇ、３，５−ジニトロサリチル酸１ｇ、亜硫酸ナトリウム０．０５ｇ、水酸化ナトリウム０．２ｇ、及び蒸留水１００ｍＬを混合することにより調製した。
(Measurement of xylanase activity)
100 μL of each sample described in Tables 1 to 3 was added to 1 mL of 1% xylan (1% xylan (derived from beech material, manufactured by SIGMA)) / 50 mM MOPS (manufactured by Nacalai Tesque), 10 mM CaCl 2 ) at pH 7.0. Each was added, vortexed, and incubated at 37 ° C. for 15 minutes. 500 μL of this was taken and put in 500 μL of a DNS solution having the following composition. After heating at 100 ° C. for 5 minutes, the mixture was allowed to cool and the absorbance at 595 nm was measured. Glucose solutions with various concentrations were prepared and reacted in the same manner with a DNS solution to create a calibration curve. From the calibration curve, the eluted reducing sugar concentration (mg / mL) was determined. The reducing sugar concentration was converted to glucose and used as the xylanase activity value. The results are shown in Tables 1-3. The DNS solution was prepared by mixing 0.2 g of phenol, 1 g of 3,5-dinitrosalicylic acid, 0.05 g of sodium sulfite, 0.2 g of sodium hydroxide, and 100 mL of distilled water.
また、表１の実施例１〜１６において、比較例１のキシラナーゼ活性を１００とした場合の相対値を併せて示す。表２の実施例１７〜３２において、比較例２のキシラナーゼ活性を１００とした場合の相対値を併せて示す。表３の実施例３３〜４８において、比較例３のキシラナーゼ活性を１００とした場合の相対値を併せて示す。 Moreover, in Examples 1-16 of Table 1, the relative value when the xylanase activity of the comparative example 1 is set to 100 is shown collectively. In Examples 17-32 of Table 2, relative values when the xylanase activity of Comparative Example 2 is taken as 100 are also shown. In Examples 33-48 of Table 3, the relative values when the xylanase activity of Comparative Example 3 is taken as 100 are also shown.
（ＣＭＣａｓｅ活性測定）
ｐＨ７．０の２％カルボキシメチルセルロース（ＣＭＣ）（２％ＣＭＣ（ＳＩＧＭＡ社製）／５０ｍＭのＭＯＰＳ（ナカライテスク社製），１０ｍＭのＣａＣｌ２）１ｍＬに表４記載の各例の試料１００μＬをそれぞれ入れ、ボルテックス後、３７℃、１５分インキュベートした。これを５００μＬずつ採り、上記と同様の組成のＤＮＳ溶液５００μＬにそれぞれ入れた。１００℃、５分間加熱後、放冷し、５９５ｎｍの吸光度を測定した。各種濃度のグルコース溶液を調製し、ＤＮＳ溶液で同様に反応させ、検量線を作成した。検量線から、溶出した還元糖濃度（ｍｇ／ｍＬ）を求めた。還元糖濃度は、グルコース換算し、これをＣＭＣａｓｅ活性値とした。結果を表４に示す。また、表４の実施例４９〜６４において、比較例４のＣＭＣａｓｅ活性を１００とした場合の相対値を併せて示す。
(Measurement of CMCase activity)
100 μL of each sample shown in Table 4 was put in 1 mL of 2% carboxymethylcellulose (CMC) (2% CMC (manufactured by SIGMA) / 50 mM MOPS (manufactured by Nacalai Tesque), 10 mM CaCl 2 ) at pH 7.0. After vortexing, the mixture was incubated at 37 ° C. for 15 minutes. 500 μL of this was taken and put in 500 μL of a DNS solution having the same composition as above. After heating at 100 ° C. for 5 minutes, the mixture was allowed to cool and the absorbance at 595 nm was measured. Glucose solutions with various concentrations were prepared and reacted in the same manner with a DNS solution to create a calibration curve. From the calibration curve, the eluted reducing sugar concentration (mg / mL) was determined. The reducing sugar concentration was converted to glucose and used as the CMCase activity value. The results are shown in Table 4. Moreover, in Examples 49-64 of Table 4, the relative value when the CMCase activity of the comparative example 4 is set to 100 is also shown.
次に、上記実施形態及び別例から把握できる技術的思想について、それらの効果とともに以下に追記する。
（ａ）有機残渣分解剤中におけるグリホサート化合物の含有量は、０．０１〜７質量％である前記有機残渣分解剤。従って、この（ａ）に記載の発明によれば、グリホサート化合物による多糖類分解酵素の活性化促進作用をより向上させることができる。
Next, technical ideas that can be grasped from the above-described embodiment and other examples will be described below together with their effects.
(A) Content of glyphosate compound in organic residue decomposing agent is said organic residue decomposing agent which is 0.01-7 mass%. Therefore, according to the invention described in (a), the activation promoting action of the polysaccharide-degrading enzyme by the glyphosate compound can be further improved.
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| JPH05170614A (en) * | 1991-05-21 | 1993-07-09 | Mycogen Corp | Herbicidal composition and method of controlling growth of plant by using same |
| JPH0892021A (en) * | 1994-09-26 | 1996-04-09 | Kao Corp | Activity-increasing agent composition for agrichemical and agrichemical composition |
| JP2001244756A (en) * | 2000-03-01 | 2001-09-07 | Nippon Columbia Co Ltd | Power amplifier |
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| JPH05170614A (en) * | 1991-05-21 | 1993-07-09 | Mycogen Corp | Herbicidal composition and method of controlling growth of plant by using same |
| JPH0892021A (en) * | 1994-09-26 | 1996-04-09 | Kao Corp | Activity-increasing agent composition for agrichemical and agrichemical composition |
| JP2001244756A (en) * | 2000-03-01 | 2001-09-07 | Nippon Columbia Co Ltd | Power amplifier |
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