JP4866689B2 - Method for saccharification of lignocellulosic plant material - Google Patents

Description

  The present invention relates to a method for saccharification of lignocellulosic plant material and a method for pulverizing lignocellulosic plant material useful as a pretreatment method for saccharification of lignocellulosic plant material. More specifically, the present invention treats lignocellulosic plant material safely and efficiently while reducing the environmental load and thermal energy and cost required for the treatment, so that sugar can be produced from the lignocellulosic plant material at a high rate. The present invention relates to a saccharification method that can be obtained and a method for pulverizing lignocellulosic plant material that can be effectively used therefor.

In recent years, consumption of fossil resources such as oil, natural gas, and coal has been increasing due to various factors such as rapid population growth and industrial development in each country. Problems such as global warming and environmental pollution caused by carbon dioxide generated are occurring on a global scale.
In order to cope with such a decrease in fossil resources and the deterioration of the global environment, it is becoming more effective to produce fuels such as ethanol using plants that can be repeatedly produced as raw materials. There is a strong demand for the development of highly functional technologies.

  As a representative example of a technique for producing a fuel using a plant as a raw material, a technique for producing a sugar from a plant and producing ethanol useful as a fuel from the obtained sugar is known. At that time, as a raw material plant for obtaining sugar, starch-based plant material made of a plant containing a high concentration of sugar such as sugar cane or a plant containing a lot of easily saccharified starch such as corn and sweet potato. Conventionally used in general. On the other hand, lignocellulosic plant materials (woody plant materials and herbaceous plant materials) containing a hardly decomposable component such as lignin are not used much. In view of this, development of an effective saccharification technique using a lignocellulosic plant material containing a hardly decomposable component such as lignin as a raw material is required.

  The technology for producing sugar from lignocellulosic materials is generally as follows: (1) acid saccharification method in which an acid such as concentrated sulfuric acid or dilute sulfuric acid is added to lignocellulosic plant material to make cellulose directly into sugar; (2) lignocellulosic plant After pre-treating the material to cause degradation and damage of plant cell walls and intercellular layers made of refractory components such as lignin, saccharification enzymes such as cellulase are added to hydrolyze cellulose into sugar. There are two types of enzymatic saccharification methods.

  In the enzymatic saccharification method of (2) above, the (2a) lignocellulosic plant material is further pulverized, steamed (steamed, crushed, steamed, hydrothermal decomposition / pressurized hot water treatment, etc.), energy rays. (2b) lignocellulosic plant material, which is treated by a physical method such as irradiation with (electron beam, γ-ray, microwave) to perform physical pretreatment to decompose and damage the cell wall of the plant. An enzymatic saccharification method that chemically decomposes and causes damage using acids (sulfuric acid, sulfurous acid, phosphoric acid), alkalis (caustic soda, ammonia, etc.), and (2c) lignocellulosic plant material It is classified as an enzymatic saccharification method in which a biological pretreatment is performed using a lignin-degrading bacterium (see Non-Patent Document 1).

  Among the above-mentioned methods, the acid saccharification method (1) and the enzymatic saccharification method (2b) that perform the chemical pretreatment are both saccharification or pretreatment of lignocellulosic plant material, such as acid and alkali. Therefore, there is a problem that the equipment cost increases because it is necessary to use equipment that can withstand such chemicals. Moreover, it is necessary to neutralize the acid or alkali used for saccharification or pretreatment and to treat a large amount of neutralized waste (for example, calcium sulfate) generated thereby, or to remove the solvent used for the pretreatment. Therefore, it takes time and cost, and there is a problem of pollution of the global environment.

  In addition, the enzymatic saccharification method in which biological pretreatment is performed using the lignin-degrading bacterium described in (2c) requires a large amount of a special bacterium for degrading lignin, and is costly. Since it takes time to decompose the lignocellulosic plant material, there is a problem that it takes a long time to decompose and saccharify the lignocellulosic plant material, and the saccharification cannot be performed efficiently.

  In the case of the enzymatic saccharification method in which the physical pretreatment of (2a) is performed, chemicals such as acid and alkali are not used for saccharification or pretreatment, so that the safety is excellent and there is little problem of pollution of the global environment. In addition, there are no problems such as post-treatment of chemicals used in the treatment and treatment of waste generated thereby. However, among the enzymatic saccharification methods in which the physical pretreatment of (2a) is performed, steaming (steaming explosion, steaming, hydrothermal decomposition, pressurized hot water treatment, etc.) or energy ray irradiation is performed as the physical pretreatment. However, it requires expensive equipment such as high-temperature and high-pressure equipment for steaming and energy beam irradiation equipment such as electron beams, γ-rays, and microwaves, which increases the equipment cost and manages the pretreatment process. It must be done strictly.

  On the other hand, among the enzymatic saccharification methods in which the physical pretreatment of the above (2a) is performed, the method of pretreating lignocellulosic plant material by pulverization and then treating with saccharifying enzyme is a special treatment such as acid or alkali. It is not necessary to use chemicals, and it is not necessary to use a steaming device (high temperature / high pressure device) or energy beam irradiation device that is expensive and requires strict management. By simply selecting and pulverizing from an appropriate pulverizer, the lignin that forms plant cells can be decomposed and damaged, and the cellulose can be easily hydrolyzed by saccharifying enzymes. It is a desirable method because it requires less equipment costs.

  Therefore, the present inventors examined a method of treating a lignocellulosic plant material by pulverization and then treating with a saccharifying enzyme among the enzymatic saccharification methods in which the physical pretreatment of (2a) is performed, It has been found that in order to make this method practical, it is necessary to increase the saccharification rate of the lignocellulosic plant material.

“Bio-energy technology and application development”, published on October 31, 2003, issued by CMC Publishing, p. 165-171

An object of the present invention is to provide a saccharification method in which lignocellulosic plant material containing a hardly degradable lignin is pulverized and powdered, and the lignocellulosic plant material powder obtained thereby is treated with a saccharifying enzyme to produce sugar. An object of the present invention is to provide a method capable of obtaining a saccharide in a higher yield than conventional methods while reducing the amount of enzyme used, shortening the treatment time with the enzyme, and lowering the treatment temperature with a simple operation.
Furthermore, the objective of this invention is providing the grinding | pulverization method of a lignocellulosic plant material which can raise the production rate of saccharide | sugar when it employ | adopts in the above-mentioned saccharification method.

The present inventors have repeatedly studied to achieve the above object. As a result, when pulverizing and pulverizing lignocellulosic plant material, when at least one component selected from wheat bran and powder is added and pulverized, plant material mainly formed from lignin is obtained. When the lignocellulosic plant material powder obtained by pulverization is treated with a saccharifying enzyme, the amount of the enzyme used is reduced, the treatment time with the enzyme is shortened, and the treatment temperature is lowered. It was found that sugar can be obtained in high yield.
Further, the inventors selected at least one component selected from wheat bran and powder and pulverized lignocellulosic plant material into powder to select from wheat bran and powder. When at least one component selected from a surfactant and a manganese salt is further added together with at least one component selected from the above, the pulverization treatment further damages and damages the cell wall of the plant material mainly formed from lignin. When the lignocellulosic plant material powder that has been promoted and obtained by the pulverization treatment is treated with a saccharifying enzyme, the sugar is further reduced while reducing the amount of enzyme used, shortening the treatment time with the enzyme, and lowering the treatment temperature. It has been found that a high yield can be obtained.

Furthermore, the present inventors pulverized lignocellulosic plant material, pulverized, and processed lignocellulosic plant material powder thus obtained with a saccharifying enzyme to produce sugar. When the saccharifying enzyme treatment is performed with at least one component selected from wheat bran and powder, the amount of the enzyme used is reduced, the treatment time with the enzyme is shortened, and the treatment temperature is lowered while the sugar is increased. It was found that it can be obtained in a yield.
In addition, the present inventors treated at least one component selected from a surfactant and a manganese salt together with at least one component selected from wheat bran and powder from the treatment of lignocellulosic plant material powder with a saccharifying enzyme. It was also found that when added, sugar can be obtained in a higher yield while reducing the amount of enzyme used, shortening the treatment time with the enzyme, and lowering the treatment temperature.

Furthermore, the present inventors have performed 100 parts by mass of lignocellulosic plant material or lignocellulosic plant material powder before pulverization by pulverizing the lignocellulosic plant material and / or treating the lignocellulosic plant material powder with a saccharifying enzyme. On the other hand, it has been found that when at least one component selected from wheat bran and powder is added at a ratio of 0.1 to 10 parts by mass, saccharification of the lignocellulosic plant material is performed well.
In addition, when the inventors further add at least one component selected from a surfactant and a manganese salt together with the wheat bran and / or powder, at least one selected from a surfactant and a manganese salt is used. When the amount of seed component added is 0.1 to 1 part by mass with respect to 100 parts by mass of lignocellulosic plant material powder, the saccharification of the lignocellulosic plant material is found to be carried out satisfactorily. Based on this, the present invention has been completed.

That is, the present invention
(1) A method for saccharification of a lignocellulosic plant material, wherein the lignocellulosic plant material is pulverized by pulverization, and the lignocellulosic plant material powder obtained thereby is treated with a saccharifying enzyme to produce sugar. One or both of pulverization treatment of cellulosic plant material and treatment of lignocellulosic plant material powder with saccharifying enzyme are performed by adding at least one component selected from wheat bran and powder. A process for saccharification of lignocellulosic plant material; and
(2) At least one component selected from a surfactant and a manganese salt is further added to the pulverization treatment of lignocellulosic plant material and / or the treatment of lignocellulose plant material powder with a saccharifying enzyme The saccharification method of (1) above,
It is.

And this invention,
(3) 0.1 to 10 parts by mass of at least one component selected from wheat bran and powder with respect to 100 parts by mass of lignocellulosic plant material before pulverization or powdered lignocellulosic plant material powder The saccharification method of (1) or (2), which is added at a ratio of:
(4) 0.1 to 1 mass of at least one component selected from a surfactant and a manganese salt with respect to 100 parts by mass of lignocellulosic plant material before pulverization or powdered lignocellulose plant material powder The saccharification method according to (2) or (3), which is added at a ratio of parts;
It is.

Furthermore, the present invention provides
(5) A method for pulverizing lignocellulosic plant material, comprising adding at least one component selected from wheat bran and powder to the lignocellulosic plant material to pulverize the lignocellulosic plant material; and ,
(6) The pulverization method according to (5), wherein at least one component selected from a surfactant and a manganese salt is further added to pulverize the lignocellulosic plant material;
It is.
And this invention,
(7) The said (5) or (6) which adds at least 1 sort (s) of component chosen from wheat bran and a powder with respect to 100 mass parts of lignocellulosic plant materials in the ratio of 0.1-10 mass parts. Grinding method; and
(8) Said (6) or (7) which adds at least 1 sort (s) of component chosen from surfactant and manganese salt with respect to 100 mass parts of lignocellulosic plant material in the ratio of 0.1-1 mass part. Crushing method of
It is.

In the case of the present invention, the lignocellulosic plant material is pretreated by a mechanical method called pulverization and then saccharified by treatment with a saccharifying enzyme. There are problems of neutralization and removal treatment of chemicals used in pretreatment, treatment of neutralization waste generated by chemical neutralization treatment, and environmental pollution caused by it, as in the method of saccharification with saccharifying enzymes after treatment. In addition, the pretreatment of lignocellulosic plant material using a steaming device (high temperature / high pressure device) or energy beam irradiation device, followed by treatment with a saccharifying enzyme and saccharification, can be used, No need for strict process control, just select and use the appropriate pulverizer among conventional pulverizers, and smooth the lignin that forms plant cells Solution (destruction) can be damaged.
And by processing with a saccharification enzyme using the lignocellulose plant material powder obtained by the said grinding | pulverization process, saccharide | sugar can be obtained smoothly with a high yield.

Further, in the case of the present invention, the lignocellulosic plant material is pulverized and pulverized, and the lignocellulosic plant material powder thus obtained is treated with a saccharifying enzyme to produce sugar. At least one of the treatment and the treatment with lignocellulosic plant material powder by saccharification enzyme is mainly formed from lignin by adding at least one component selected from wheat bran and powder, which are readily available. When the lignocellulosic plant material powder obtained by the pulverization treatment is treated with a saccharifying enzyme, the amount of the enzyme used is reduced and the treatment time of the enzyme is reduced. Sugar can be obtained in a higher yield than before, while shortening and lowering the treatment temperature.
In the present invention, at least one of pulverization treatment of lignocellulosic plant material and treatment of lignocellulosic plant material powder with a saccharifying enzyme, together with at least one component selected from wheat bran and powder, surfactant and manganese salt When at least one component selected from the above is further added, a sugar can be obtained in a higher yield than before.
In the case of the method for pulverizing lignocellulosic plant material of the present invention, lignocellulosic plant material powder that can be effectively used for saccharification treatment with a saccharifying enzyme can be produced smoothly by a simple operation. The lignocellulose plant material powder obtained by the pulverization method of the present invention is useful not only for saccharification treatment with a saccharifying enzyme but also for producing other substances using lignocellulose plant material as a raw material.

The present invention is described in detail below.
The lignocellulosic plant material used in the present invention may be any plant material mainly composed of lignin and cellulose.
Examples of the lignocellulosic plant material used in the present invention include woody materials (tree trunks, branches, roots, leaves, etc.), herbaceous materials (rice, wheat, corn, sugarcane, beans, etc.) and weed stalks. And one or more of these can be used.

  The lignocellulosic plant material is preferably used in a dry state or a state close thereto so that the pulverization can be carried out smoothly. Generally, the lignocellulosic plant material can be subjected to a pulverization treatment with a water content of 20% by mass or less. preferable. The lignocellulosic plant material is preferably preliminarily formed into chips having a size of 5 cm or less, more preferably 2 cm or less, and particularly 1 cmm or less in order to facilitate pulverization.

Lignocellulosic plant materials usually contain a large amount of lignin in the cell walls and intercellular layers constituting the plant, and a cellulose-rich layer exists inside the cell wall containing a large amount of lignin. Lignin contained in large amounts in the cell walls and intercellular layers of plant cells is hardly degradable, and even if lignocellulosic plant material is treated with saccharifying enzymes as they are, it is difficult for saccharifying enzymes to reach the cellulose in the cell walls. Is difficult to disassemble.
In the present invention, by pulverizing lignocellulosic plant material, cell walls and intercellular layers containing a large amount of lignin are destroyed, decomposed, damaged, etc., and holes are made in the cell wall, or the cellulose layer in the cell wall As a result, the saccharification treatment by the next saccharifying enzyme is facilitated.

  In pulverization of lignocellulosic plant material, any of a pulverization method and a pulverizer capable of pulverizing lignocellulosic plant material can be adopted. It is preferably employed because it can effectively destroy, decompose, damage, etc. the cell walls and intercellular layers of the material. Examples of the pulverizing apparatus suitably used for pulverizing lignocellulosic plant material include a ball mill.

The lignocellulosic plant material is preferably pulverized so as to obtain a powder having an average particle size of 500 μm or less, more preferably 200 μm or less, and particularly 100 μm or less. If the average particle size of the lignocellulosic plant material powder after pulverization is too large, the cell walls and intercellular layers of the lignocellulosic plant material are not sufficiently destroyed, decomposed, or damaged, and the treatment with the next saccharifying enzyme is sufficient. It becomes difficult to be done. The lower limit value of the average particle size of the lignocellulosic plant material powder after pulverization is not particularly limited, but if it is too small, the pulverization requires a long time and high cost, so the average particle size is preferably 10 μm or more.
The lignocellulosic plant material powder pulverized by the method of the present invention can be effectively used in a saccharification method using a saccharification enzyme. In addition to saccharification with saccharifying enzymes, lignocellulosic plant material powders obtained by pulverization can be used for other applications for producing various compounds and products using lignocellulosic plant materials as raw materials (for example, methane, acetone, ethanol). And alcohols such as butanol, and organic acids such as lactic acid).

The present invention includes various embodiments such as the following (I) to (XII).
(I) After at least one component selected from wheat bran and powder (hereinafter sometimes abbreviated as “wheat bran etc.”) is added to the lignocellulosic plant material, The saccharification method which manufactures saccharide | sugar by processing the lignocellulose plant material powder obtained by this with a saccharification enzyme.
(II) A saccharification method in which lignocellulosic plant material is ground and then wheat bran or the like is added to the resulting lignocellulosic plant material powder and treated with a saccharifying enzyme to produce sugar.
(III) After adding a part of wheat bran and crushing lignocellulosic plant material, add the rest of wheat bran to the resulting lignocellulosic plant material powder and treating with saccharifying enzyme Saccharification method for producing sugar.

(IV) After adding a wheat bran and at least one component selected from a surfactant and a manganese salt to pulverize the lignocellulosic plant material, the lignocellulosic plant material powder obtained thereby is obtained. A saccharification method in which sugar is produced by treatment with a saccharifying enzyme.
(V) After adding wheat bran or the like and grinding the lignocellulosic plant material, add at least one component selected from surfactants and manganese salts to the lignocellulosic plant material powder obtained thereby. And then saccharifying by treating with a saccharifying enzyme.
(VI) After pulverizing lignocellulosic plant material, to the lignocellulosic plant material powder obtained by adding wheat bran, etc., at least one component selected from surfactants and manganese salts, A saccharification method for producing sugar by treatment.
(VII) Adding at least one component selected from a surfactant and a manganese salt to pulverize the lignocellulosic plant material, and adding wheat bran or the like to the resulting lignocellulosic plant material powder A saccharification method in which sugar is produced by treatment with a saccharifying enzyme.

(VIII) After adding a part of wheat bran and pulverizing the lignocellulosic plant material, the lignocellulosic plant material powder obtained from the remainder of the wheat bran and the surfactant and manganese salt A saccharification method in which at least one selected component is added and treated with a saccharifying enzyme to produce sugar.
(IX) After adding a part of at least one component selected from a surfactant and a manganese salt to pulverize the lignocellulosic plant material, wheat liquor is added to the lignocellulosic plant material powder obtained thereby. And the like, and a saccharification method in which the remainder of at least one component selected from a surfactant and a manganese salt is added and treated with a saccharifying enzyme to produce sugar.
(X) Lignocellulose obtained by adding a part of wheat bran and a part of at least one component selected from a surfactant and a manganese salt to pulverize the lignocellulosic plant material. A saccharification method wherein sugar is produced by adding a remainder of wheat bran and the like and a remainder of at least one component selected from a surfactant and a manganese salt to a plant material powder and treating with a saccharifying enzyme.

(XI) A method of pulverizing lignocellulosic plant material by adding wheat bran or the like.
(XII) A method of pulverizing lignocellulosic plant material by adding wheat bran or the like and at least one component selected from surfactants and manganese salts.

  Among the various aspects described above, the above-described (I), (III), (IV), (V), (VII), and (X) are added with at least wheat bran and the like at the time of pulverizing the lignocellulosic plant material. , (XI) and (XII), among which the methods (IV) and (V) are pulverized by further promoting the destruction, decomposition, damage, etc. of persistent lignin in lignocellulosic plant materials. When lignocellulosic plant material powder obtained by the above is treated with a saccharifying enzyme, the saccharification rate can be further increased and the operation is simple, and thus it is preferably employed.

  In the present invention, only one of wheat bran and powder may be used, or both may be used in combination. Among them, it is preferable to use wheat bran from the viewpoints of cost, availability, and the like.

Addition amount of wheat bran, etc. (If both wheat bran and powder are added, the total addition amount, and if adding wheat bran, etc. in both the grinding and saccharification treatment steps, the sum of both treatment steps) The addition amount) is preferably 0.1 to 10 parts by mass, and 1 to 8 parts by mass with respect to 100 parts by mass of the lignocellulose plant material before pulverization or the lignocellulose plant material powder after pulverization. It is more preferable, and it is still more preferable that it is 2-5 mass parts.
If the amount of wheat bran added is too small, the effect of improving the saccharification rate is insufficient. On the other hand, if the amount is too large, a large amount of saccharification enzyme is consumed for saccharification of wheat bran added in a large amount. It becomes easy to spoil the original purpose of saccharification of the material.

  In the present invention, from the viewpoint that the saccharification rate becomes higher, either or both of the lignocellulosic plant material powder obtained by crushing and crushing the lignocellulosic plant material powder with the saccharifying enzyme are combined with wheat bran and the like. It is preferable to carry out by adding either one or both of a surfactant and a manganese salt, and more preferably by adding both a surfactant and a manganese salt.

As the surfactant used in that case, any of an anionic surfactant, a cationic surfactant and a nonionic surfactant can be used. Examples of anionic surfactants include higher fatty acid alkali salts, alkyl sulfates, alkyl sulfonates, alkyl aryl sulfone salts, sulfosuccinic acid ester salts, etc. Examples of cationic surfactants Can include higher amine halides, alkylpyridinium halides, quaternary ammonium surfactants, and examples of nonionic surfactants include polyethylene glycol alkyl ethers, polyethylene glycol fatty acid esters. , Sorbitan fatty acid ester, fatty acid monoglyceride and the like, and one or more of these can be used.
Among them, nonionic surfactants are preferably used from the viewpoint that they are easily used in combination with other surfactants and are not easily affected by electrolytes, and specific examples thereof include polyoxyethylene sorbitan monolaurate. Mention may be made of polyoxyethylene sorbitan fatty acid esters.

As the manganese salt, either a manganese salt of an inorganic acid or a manganese salt of an organic acid can be used. Specific examples include manganese salts of inorganic acids such as manganese sulfate and manganese chloride, and manganese salts of organic acids such as manganese acetate. One or two or more of these can be used.
Among them, in the present invention, manganese sulfate is preferably used from the viewpoints of price and storage stability.

When adding at least one component selected from a surfactant and a manganese salt together with wheat bran and the like, the addition amount of at least one component selected from a surfactant and a manganese salt (when using two or more components) Is the total addition amount, and when added in both the pulverization and saccharification treatment steps, the total addition amount in both treatment steps) is the lignocellulosic plant material before pulverization or the lignocellulose plant material powder after pulverization It is preferable that it is 0.1-1 mass part with respect to 100 mass parts, It is more preferable that it is 0.1-0.8 mass part, It is further that it is 0.1-0.5 mass part preferable.
An excessive amount of at least one component selected from a surfactant and a manganese salt is not preferable because the reaction is hindered.

  Wheat bran and at least one component selected from surfactants and manganese salts are simultaneously added to lignocellulosic plant material (lignocellulose plant material before pulverization and / or lignocellulosic plant material after pulverization). In this case, wheat bran or the like and at least one component selected from surfactants and manganese salts may be added separately to the lignocellulosic plant material, or the wheat bran and the like, surfactant and manganese. At least one component selected from salts may be mixed in advance, and the mixture may be added to the lignocellulosic plant material.

  In the present invention, the pulverized lignocellulosic plant material may be treated with a saccharifying enzyme following the pulverization of the lignocellulosic plant material, or the lignocellulosic plant material powder may be produced and stored in advance by pulverization. In addition, the lignocellulosic plant material powder may be used in the required amount when necessary, and saccharification treatment may be performed, or lignocellulosic plant material powder may be purchased or obtained separately and used for saccharification treatment. May be.

The saccharification treatment of lignocellulosic plant material powder with a saccharifying enzyme can be carried out in the same manner as a conventionally known saccharification method in which a saccharification enzyme is allowed to act on lignocellulose plant material powder.
As the saccharification enzyme used for the saccharification treatment of the lignocellulosic plant material powder, the same saccharification enzyme conventionally used in the saccharification treatment of lignocellulose plant material can be used, and representative examples thereof include cellulase, A hemicellulase etc. can be mentioned, The mixture of these enzymes is used preferably.
Various saccharifying enzymes are already commercially available. Specific examples include, for example, “Onozuka” manufactured by Yakult Pharmaceutical Co., Ltd., “Meiselase” manufactured by Meiji Seika Co., Ltd., and commercially available products are purchased and used. be able to. Many commercially available saccharifying enzymes generally contain both cellulase and hemicellulase.

The amount of saccharifying enzyme used may vary depending on the treatment conditions during saccharification, the type of lignocellulosic plant material, the particle size of the lignocellulosic plant material powder, the cellulose content, etc., but in general lignocellulosic plants From 1 g of material powder, it is preferable that it is 1000 units or less, Furthermore, it is 10-500 units, especially 100-300 units from the point which performs saccharification smoothly.
In addition, the “unit” of the cellulase in this specification is FPU (Filter Paper Unit), and is defined by 1 FPU = 1 μmol Glucose / min.

The saccharification treatment can be carried out according to the methods and conditions conventionally employed when sugar is produced by allowing a saccharifying enzyme to act on lignocellulosic plant material powder.
Although it is not limited, for example, lignocellulosic plant material powder containing wheat bran or the like, or lignocellulosic plant material containing at least one component selected from wheat bran and the like, a surfactant and a manganese salt Suspend the powder in an aqueous solution adjusted to a pH at which the saccharifying enzyme works smoothly (usually pH 4-5), or suspend the lignocellulosic plant material powder added with wheat bran or the like in the aqueous solution adjusted to the above pH. At least one component selected from a surfactant and a manganese salt is added to the suspension and suspended, or selected from a lignocellulosic plant material powder, wheat bran, a surfactant, and a manganese salt. At least one component is individually suspended in an aqueous solution adjusted to the above pH, and saccharifying enzyme is added thereto at the above ratio. The sugar contained in the lignocellulose plant material is hydrolyzed to generate sugar by hydrolyzing the mixture at a temperature of 30 to 80 ° C. for about 0.5 to 3 days with stirring. Can do.
The amount of suspension of the lignocellulosic plant material powder in the aqueous solution is generally about 0.1 to 20% by mass, particularly about 0.5 to 15% by mass based on the mass of the aqueous solution, and the reaction proceeds smoothly. From the point of view, it is preferable.

A reaction liquid containing sugar is generated by the above-described saccharification treatment. The reaction solution containing sugar obtained in this manner may be used as it is or in a subsequent reaction such as alcoholic fermentation (production of ethanol) while leaving an unreacted reaction residue (solid residue) in a liquid state. Alternatively, the solid content (solid residue) may be separated and removed from the reaction solution by an appropriate means such as filtration to collect an aqueous solution, and the aqueous solution may be dried to recover the sugar. The recovered sugar may be used as it is, or may be used as a raw material for producing alcohol or other compounds.
When alcohol fermentation is performed using the reaction solution containing sugar obtained above, it can be performed in the same manner as a conventionally known method, and is not limited at all.
Moreover, in this invention, you may make it perform alcohol fermentation simultaneously at the time of the saccharification process by a saccharification enzyme as needed.

At the time of pulverization treatment of lignocellulosic plant material and / or saccharification treatment of lignocellulosic plant material powder, wheat bran or the like is added, or at least one component selected from wheat bran and the like, surfactant and manganese salt The reason for improving the saccharification rate when lignocellulosic plant material is treated with a saccharifying enzyme is not clear, but is presumed as follows.
That is, at least one component selected from any active ingredient and / or surfactant and manganese salt contained in wheat bran or the like, when pulverizing lignocellulosic plant material, such as lignin in lignocellulosic plant material Any active ingredient that acts directly on the hardly decomposable component to promote the destruction, decomposition, damage, perforation, etc. of the cell wall and intercellular layer containing a large amount of the hardly decomposable component, and is also contained in wheat bran during the saccharification reaction And / or at least one component selected from a surfactant and a manganese salt further promotes destruction, decomposition, damage, etc. of cell walls and intercellular layers that have been decomposed or damaged by crushing, whereby It is presumed that the saccharification rate is improved because the cellulose layer is easily reached.

  EXAMPLES The present invention will be specifically described below with reference to examples, but the present invention is not limited to the examples below.

Example 1
(1) Place 15 alumina balls (diameter 2 cm) in a ball mill (FRITSH “P-5”), and add cedar chips (diameter 1 cm), wheat bran (Nisshin Flour Milling Co., Ltd.), interface Activator [“TWEEN 20” manufactured by Kanto Chemical Co., Inc. (nonionic surfactant made of polyoxyethylene sorbitan monolaurate)] and manganese sulfate were added in the amount (g) shown in Table 1 below, and under the condition of 300 rpm. The powder was pulverized for 2 hours.
(2) 500 U of the powder obtained in (1) above is suspended in 50 ml of Britton-Robinson buffer (pH 4.5), and 180 U (unit) of saccharifying enzyme (“Onozuka P-1500” manufactured by Yakult Pharmaceutical Co., Ltd.) ) Was added, and the enzyme reaction was carried out by shaking at 40 ° C. for 45 hours.
(3) After the reaction of (2) above, the liquid containing the saccharified product was separated by filtration, the solid residue was dried at 135 ° C. for 2 hours, and its mass (A ₁ ) (mg) was measured.
(4) Also, in order to determine the saccharification rate of cedar powder, 15 balls made of alumina (diameter 2 cm) were put in the same ball mill used in (1) above, and used in (1) above. Only 5 g of the same wheat bran was added and pulverized for 2 hours under the same conditions as in (1) above, and 500 mg of the wheat bran powder thus obtained was suspended in 50 ml of the same buffer used in (2) above. After adding 180 U (unit) of the same saccharifying enzyme as used in (2) above, the enzyme reaction was performed by shaking at 40 ° C. for 45 hours. After the reaction, the liquid containing the saccharified product was separated by filtration, the solid residue was dried at 135 ° C. for 2 hours, and the mass was measured.
(5) Based on the results of (3) and (4) above, the saccharification rate of cedar was determined by the following mathematical formula (1).

Cedar saccharification rate (% by mass) = 100 − {(A ₁ −B ₁ ) / (A ₀ −B ₀ )} × 100 (1)
Where
A ₀ = mass of cedar in the powder (500 mg) used in the saccharification treatment of (2) above (mg) = 500 mg × [5 g ÷ {(mass of cedar chips used during pulverization) (5 g) + (wheat bran Mass) (0 g or 0.25 g) + (total mass of surfactant and manganese sulfate) (0 g, 0.01 g, 0.015 g or 0.025 g)}]
A ₁ = mass (mg) after drying of the solid residue after saccharification treatment obtained in (3) above
B ₀ = mass of wheat bran in the powder (500 mg) used for the saccharification treatment of (2) above (mg) = 500 mg × [(0 g or 0.25 g) ÷ {(mass of cedar chips used during grinding) ( 5 g) + (mass of wheat bran) (0 g or 0.25 g) + (total mass of surfactant and manganese sulfate) (0 g, 0.01 g, 0.015 g or 0.025 g)}]
B ₁ = mass of dry solid residue derived from wheat bran contained in the dry solid residue after saccharification obtained in (3) above (mg) = {after saccharification of wheat bran measured in (4) above Mass after drying of solid residue (mg)} × [(0 g or 0.25 g) ÷ {(mass of cedar chips used during pulverization) (5 g) + (mass of wheat bran) (0 g or 0.25 g) + (Total weight of surfactant and manganese sulfate) (0 g, 0.01 g, 0.015 g or 0.025 g)}]

As seen in the results of Table 1 above, in Experiment Nos. 2-5, wheat bran was added, or cedar chips were ground by adding wheat bran and surfactant and / or manganese sulfate, thereby By saccharifying the resulting powder with a saccharification enzyme, the saccharification rates of cedar (lignocellulose plant material) were 35.6% by weight, 40.8% by weight, 45.6% by weight and 49.8%, respectively. The value was as high as mass%.
On the other hand, in Experiment No. 1, cedar chips were crushed without adding any of wheat bran, surfactant and manganese sulfate, and saccharification treatment was carried out with the saccharification enzyme as it was using the resulting powder. Therefore, the saccharification rate was as low as 26.5% by mass.

Example 2
(1) Place 15 alumina balls (diameter 2 cm) in a ball mill (FRITSH "P-5 type"), and put 5 g of cedar chips (diameter 1 cm) in this, 2 hours at 300 rpm. The powder was pulverized.
(2) To the powder obtained in the above (1), wheat bran (manufactured by Nisshin Flour Milling Co., Ltd.) and a surfactant [“TWEEN20” (manufactured by Kanto Chemical Co., Ltd.) (nonionic surfactant comprising polyoxyethylene sorbitan monolaurate) Agent)] in the amount shown in Table 2 below to prepare a mixture, and 500 mg of the resulting mixture is suspended in 50 ml of Briton-Robinson buffer (pH 4.5) and saccharifying enzymes (cellulase, Yakult) 180 U of Yakuhin Kogyo Co., Ltd. “Onozuka P-1500”) was added, and the mixture was shaken at 40 ° C. for 45 hours to carry out the enzyme reaction.
(3) After the reaction of (2) above, the liquid containing the saccharified product was separated by filtration, the solid residue was dried at 135 ° C. for 2 hours, and its mass (C ₁ ) (mg) was measured.
(4) In order to determine the saccharification rate of cedar, 500 mg of the same wheat bran powder used in (2) above was suspended in 50 ml of the same buffer used in (2) above, and (2 After adding 180 U of the same saccharifying enzyme used in the above, the enzyme reaction was performed by shaking at 40 ° C. for 45 hours. After the reaction, the liquid containing the saccharified product was separated by filtration, the solid residue was dried at 135 ° C. for 2 hours, and the mass was measured.
(5) Based on the results of (3) and (4) above, the saccharification rate of cedar was determined by the following mathematical formula (2).

Cedar saccharification rate (% by mass) = 100 − {(C ₁ −D) / C ₀ } × 100 (2)
Where
C ₀ = mass of cedar powder used in the saccharification treatment of (2) above (500 mg)
C ₁ = mass (mg) after drying of the solid residue after saccharification obtained in (3) above
D = mass of dry solid residue derived from wheat bran contained in dry solid residue after saccharification obtained in (3) above (mg) = {after saccharification of wheat bran measured in (4) above Mass after drying solid residue (mg)} × [(0 g or 0.25 g) ÷ {mass of cedar powder used during saccharification treatment (5 g) + mass of wheat bran (0 g or 0.25 g) + surfactant Mass (0 g or 0.015 g)}]

As shown in the results of Table 2 above, in Experiment No. 6, when cedar powder obtained by pulverizing cedar chips was saccharified with a saccharifying enzyme, saccharification was performed by adding wheat bran and performing saccharification treatment. The saccharification rate was 30.3% by mass, and the saccharification rate was about 10% compared to Experiment No. 1 (saccharification rate 26.5% by mass) in which cedar powder was saccharified with a saccharifying enzyme without adding wheat bran and surfactant. It is 4 mass% higher.
In Experiment No. 7, when cedar powder obtained by pulverizing cedar chips was saccharified with a saccharifying enzyme, saccharification rate of 40 cedar was obtained by adding saccharified wheat flour and a surfactant. Compared to Experiment No. 1 (saccharification rate 26.5% by mass) in which cedar powder was saccharified with saccharification enzyme without adding wheat bran and surfactant, the saccharification rate was about 14% by mass. Is also high.

Example 3
(1) Put 15 alumina balls (diameter 2cm) into a ball mill (FRITSH "P-5 type") and put cedar chips (diameter 1cm) and wheat bran (Nisshin Flour Milling Co., Ltd.) Were added in the amounts shown in Table 3, and pulverized for 2 hours under the condition of 300 rpm to obtain powder.
(2) Surfactant [“TWEEN 20” (non-ionic surfactant made of polyoxyethylene sorbitan monolaurate)] manufactured by Kanto Chemical Co., Inc.) with respect to the mass of the powder obtained in (1) above is shown in Table 3 below. As shown in Fig. 3, the mixture was mixed at 0.3% by mass (extra), and 500 mg of the resulting mixture was suspended in 50 ml of Briton-Robinson buffer (pH 4.5), and saccharifying enzyme (cellulase, Yakult Pharmaceutical Co., Ltd.) After adding 180 U of “Onozuka P-1500” manufactured by the company, the enzyme reaction was performed by shaking at 40 ° C. for 45 hours.
(3) After the reaction of (2) above, the liquid containing the saccharified product is separated by filtration, the solid residue is dried at 135 ° C. for 2 hours, and its mass (A ₁ ) (mg) is measured. When the saccharification rate of cedar was determined by (1), it was as shown in Table 3 below.

  As seen in the results of Table 3 above, in Experiment No. 8, wheat bran was added and cedar chips were pulverized, and a surfactant was further added to the resulting powder and saccharified with a saccharifying enzyme. As a result, the saccharification rate of cedar was as high as 48.9% by mass, and the saccharification enzyme was directly used for the saccharification treatment using the powder obtained by pulverizing cedar chips without adding wheat bran and surfactant. Compared to 1 (saccharification rate 26.5% by mass), the saccharification rate is as high as 22% by mass or more.

  In the case of the method of the present invention, lignocellulosic plant materials are conventionally pulverized without using chemicals such as acids and alkalis, steaming equipment (high temperature / high pressure equipment) and energy beam irradiation equipment with high equipment costs. The lignocellulosic material can be obtained in a high yield by pulverizing easily and smoothly using an apparatus and treating with a saccharifying enzyme using the resulting lignocellulosic plant material powder. It is highly useful as a technique for producing sugar from plant materials and as a technique for grinding lignocellulosic plant materials.

Claims (8)

  A method for saccharification of a lignocellulosic plant material, in which lignocellulosic plant material is pulverized and pulverized, and the lignocellulosic plant material powder obtained thereby is treated with a saccharifying enzyme to produce sugar. Either or both of pulverization treatment of material and treatment of lignocellulosic plant material powder with saccharification enzyme are performed by adding at least one component selected from wheat bran and powder. A method for saccharification of cellulosic plant material.   Either or both of the pulverization treatment of lignocellulosic plant material and the treatment of lignocellulosic plant material powder with saccharifying enzyme are performed by further adding at least one component selected from a surfactant and a manganese salt. The saccharification method according to claim 1.   100 parts by mass of lignocellulosic plant material before pulverization or powdered lignocellulosic plant material powder, at a ratio of 0.1 to 10 parts by mass of at least one component selected from wheat bran and powder. The saccharification method according to claim 1 or 2, which is added.   A ratio of 0.1 to 1 part by mass of at least one component selected from a surfactant and a manganese salt with respect to 100 parts by mass of the lignocellulosic plant material or pulverized lignocellulosic plant material powder before pulverization The saccharification method according to claim 2 or 3, which is added in the step.   A method for pulverizing a lignocellulosic plant material, comprising adding at least one component selected from wheat bran and powder to the lignocellulosic plant material to pulverize the lignocellulosic plant material.   The pulverization method according to claim 5, wherein at least one component selected from a surfactant and a manganese salt is further added to pulverize the lignocellulosic plant material.   The pulverization method according to claim 5 or 6, wherein at least one component selected from wheat bran and powder is added at a ratio of 0.1 to 10 parts by mass with respect to 100 parts by mass of the lignocellulosic plant material.   The pulverization method according to claim 6 or 7, wherein at least one component selected from a surfactant and a manganese salt is added at a ratio of 0.1 to 1 part by mass with respect to 100 parts by mass of the lignocellulosic plant material.