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WO2011099558A1 - Procédé de production de sucre, procédé de production d'éthanol et procédé de production d'acide lactique - Google Patents

Procédé de production de sucre, procédé de production d'éthanol et procédé de production d'acide lactique Download PDF

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Publication number
WO2011099558A1
WO2011099558A1 PCT/JP2011/052861 JP2011052861W WO2011099558A1 WO 2011099558 A1 WO2011099558 A1 WO 2011099558A1 JP 2011052861 W JP2011052861 W JP 2011052861W WO 2011099558 A1 WO2011099558 A1 WO 2011099558A1
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Prior art keywords
raw material
ammonia
sugar
biomass
biomass raw
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PCT/JP2011/052861
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English (en)
Japanese (ja)
Inventor
圭日子 五十嵐
昌久 和田
正浩 鮫島
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国立大学法人東京大学
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Publication of WO2011099558A1 publication Critical patent/WO2011099558A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
    • C12P7/06—Ethanol, i.e. non-beverage
    • C12P7/08—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate
    • C12P7/10—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate substrate containing cellulosic material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/56—Lactic acid
    • C—CHEMISTRY; METALLURGY
    • C13—SUGAR INDUSTRY
    • C13K—SACCHARIDES OBTAINED FROM NATURAL SOURCES OR BY HYDROLYSIS OF NATURALLY OCCURRING DISACCHARIDES, OLIGOSACCHARIDES OR POLYSACCHARIDES
    • C13K1/00—Glucose; Glucose-containing syrups
    • C13K1/02—Glucose; Glucose-containing syrups obtained by saccharification of cellulosic materials
    • 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
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00—Technologies for the production of fuel of non-fossil origin
    • Y02E50/10—Biofuels, e.g. bio-diesel

Definitions

  • the present invention relates to a method for producing sugar, a method for producing ethanol, and a method for producing lactic acid using biomass raw materials.
  • ethanol has been widely used as an alternative fuel for fossil fuels to produce ethanol from plant resources containing cellulose such as woody biomass and herbaceous biomass, and to use it as various fuels and chemical raw materials. ing.
  • Production of ethanol from biomass raw material can be performed, for example, by decomposing the collected biomass raw material into sugar in the saccharification step and then converting it to ethanol using a microorganism such as yeast in the fermentation step.
  • a microorganism such as yeast
  • the use of biodegradable polymers is increasing from the viewpoint of reducing environmental impact, and lactic acid is used as one of the raw materials. This lactic acid can also be obtained by saccharifying the biomass raw material and further fermenting it.
  • the saccharification has often been carried out using strong acids such as concentrated sulfuric acid and concentrated hydrochloric acid, but it is desired to reduce the amount of these strong acids used from the viewpoint of reducing environmental burden. Therefore, in recent years, saccharification of biomass raw materials using enzymes has been widely studied as a means to replace saccharification with strong acids such as concentrated sulfuric acid and concentrated hydrochloric acid. Enzymatic saccharification is a desirable means from the viewpoint of the influence on the environment. However, for this enzymatic saccharification, it is necessary to pre-treat biomass raw material in advance for the purpose of making the enzyme act easily.
  • the amount of cellulase added to saccharify cellulose is significantly larger than the amount of amylase added for the enzymatic saccharification of starch that has already been put to practical use, and an increase in the sugar conversion cost is a problem. .
  • the present invention makes it a subject to solve the said various problems in the past and to achieve the following objectives. That is, the present invention can efficiently carry out enzymatic saccharification, and therefore can improve sugar production efficiency, ethanol production efficiency, and lactic acid production efficiency. It is an object to provide a method and a method for producing lactic acid.
  • cellulose having a lower crystal density than cellulose type I which is natural type cellulose (for example, cellulose III type I ) as an object of enzymatic saccharification.
  • the biomass raw material is treated with a treatment agent containing at least one of ammonia and organic amine, further pulverized, and then enzymatically saccharified using cellulase and hemicellulase, thereby significantly improving the enzymatic saccharification efficiency of the biomass raw material. It is a new finding by the present inventors that it has not been known so far.
  • the present invention is based on the above findings by the present inventors, and means for solving the above problems are as follows. That is, ⁇ 1> (A) A process for obtaining a modified biomass raw material by treating a biomass raw material containing cellulose type I with a treating agent containing at least one of ammonia and an organic amine, (B) obtaining the enzyme saccharification raw material by pulverizing the modified biomass raw material; and (C) a step of enzymatically saccharifying the raw material for enzymatic saccharification using cellulase and hemicellulase to obtain a sugar; It is a manufacturing method of the sugar characterized by including.
  • ⁇ 2> The sugar production method according to ⁇ 1>, wherein the treatment agent used in the step (A) is ammonia.
  • the biomass raw material containing cellulose type I is at least one of woody biomass and herbaceous biomass.
  • ⁇ 4> The method for producing a sugar according to any one of ⁇ 1> to ⁇ 3>, wherein the hemicellulase is xylanase.
  • ⁇ 5> The sugar production method according to any one of ⁇ 1> to ⁇ 4>, wherein the mass ratio of hemicellulase to cellulase (hemicellulase / cellulase) is 0.001 to 0.5.
  • ⁇ 6> The method for producing a saccharide according to any one of ⁇ 1> to ⁇ 5>, wherein in the step (C), cellobiase is further used for enzymatic saccharification.
  • ⁇ 7> A method for producing ethanol, wherein the sugar obtained by the method for producing sugar according to any one of ⁇ 1> to ⁇ 6> is fermented to obtain ethanol.
  • ⁇ 8> A method for producing lactic acid, wherein the saccharide obtained by the method for producing saccharide according to any one of ⁇ 1> to ⁇ 6> is fermented to obtain lactic acid.
  • the conventional problems can be solved, the object can be achieved, and enzymatic saccharification can be efficiently performed. Therefore, sugar production efficiency, ethanol production efficiency, and lactic acid production A sugar production method, ethanol production method, and lactic acid production method capable of improving production efficiency can be provided.
  • FIG. 1 shows the amount of glucose produced by enzymatic saccharification using hemicellulase and cellulase after treating eucalyptus with ammonia ( ⁇ : Example 1), and enzymatic saccharification using hemicellulase and cellulase without treating eucalyptus with ammonia. It is a figure which shows the amount of glucose produced by doing ( ⁇ : Comparative Example 1-2).
  • the vertical axis represents the amount of glucose (g) obtained from 1 kg of biomass material, and the horizontal axis represents the mass ratio of hemicellulase to cellulase (hemicellulase / cellulase).
  • FIG. 2 shows the amount of glucose produced by enzymatic saccharification using hemicellulase and cellulase after treatment with Ezo noki willow ( ⁇ : Example 2), and enzymatic saccharification using Emino willow without treatment with ammonia and hemicellulase and cellulase It is a figure which shows the amount of glucose produced by doing ( ⁇ : Comparative Example 2-2).
  • the vertical axis represents the amount of glucose (g) obtained from 1 kg of biomass material, and the horizontal axis represents the mass ratio of hemicellulase to cellulase (hemicellulase / cellulase).
  • FIG. 3 shows the amount of glucose produced by enzymatic saccharification using hemicellulase and cellulase after sugar treatment of sugarcane bagasse ( ⁇ : Example 3), and sugarcane bagasse was treated with hemicellulase and cellulase without ammonia treatment. It is a figure which shows the amount of glucose produced by enzymatic saccharification ( ⁇ : Comparative Example 3-2).
  • the vertical axis represents the amount of glucose (g) obtained from 1 kg of biomass material, and the horizontal axis represents the mass ratio of hemicellulase to cellulase (hemicellulase / cellulase).
  • Example 4 shows the amount of glucose produced by enzymatic saccharification using hemicellulase and cellulase after the switch glass is treated with ammonia ( ⁇ : Example 4), and the switch glass is treated with hemicellulase and cellulase without ammonia treatment. It is a figure which shows the amount of glucose produced by enzymatic saccharification ( ⁇ : Comparative Example 4-2).
  • the vertical axis represents the amount of glucose (g) obtained from 1 kg of biomass material, and the horizontal axis represents the mass ratio of hemicellulase to cellulase (hemicellulase / cellulase).
  • the method for producing a sugar of the present invention comprises (A) a step of obtaining a modified biomass raw material by treating a biomass raw material containing cellulose type I with a treating agent containing at least one of ammonia and an organic amine, (B) Crushing the modified biomass raw material to obtain a raw material for enzymatic saccharification, and (C) enzymatic saccharification of the raw material for enzymatic saccharification using cellulase and hemicellulase to obtain a sugar, If necessary, other steps are further included.
  • the step (A) is a step of obtaining a modified biomass raw material by treating a biomass raw material containing cellulose type I with a treating agent containing at least one of ammonia and an organic amine.
  • biomass raw material containing cellulose type I is not particularly limited and can be appropriately selected according to the purpose.
  • the “waste biomass” is not particularly limited and can be appropriately selected according to the purpose.
  • black liquor discharged in the process of papermaking industrial waste such as chip waste, agriculture and forestry Examples include waste discharged during the livestock industry, by-products such as fir shells and cow dung, and general waste such as garbage and waste cooking oil.
  • the “resource crop-based biomass” is not particularly limited and may be appropriately selected depending on the purpose.
  • sugar or starch crops cultivated as food such as sugar cane and corn
  • herbaceous biomass such as woody biomass, switchgrass, napiergrass, Eliansus, Miscanthus, Susuki, sugarcane bagasse, sorghum bagasse, rice straw, wheat straw, barley, johnsongrass, and corn stover.
  • the state of the biomass raw material containing cellulose type I is not particularly limited and can be appropriately selected according to the purpose and collected. May be used as they are, or may be used in a state where the size is appropriately reduced to a certain extent by cutting, pulverization, or the like. Among these, the cut and pulverized state is preferable.
  • the size of the biomass raw material is not particularly limited and may be appropriately selected depending on the purpose.
  • the mesh opening size is preferably 5 mm or less, more preferably 3 mm or less, and even more preferably 2 mm or less. .
  • the size of the mesh opening is within the further preferable range, it is advantageous in that the processing time can be shortened and the amount of the processing agent to be used can be reduced.
  • the process of cutting and pulverizing the collected biomass material may be referred to as “coarse pulverization”.
  • the treatment with the treatment agent containing at least one of ammonia and organic amine proceeds efficiently, and when the modified biomass raw material is pulverized, it is excellent in finer and enzymatic saccharification efficiency.
  • a raw material for enzyme saccharification in the form of fine powder can be obtained efficiently.
  • a grinder used for the said rough crushing According to the objective, it can select suitably, For example, a wheelie mill, a cutter mill, a hammer mill, a pin mill etc. are mentioned.
  • the method is not particularly limited and can be appropriately selected depending on the purpose. Examples include a method in which the biomass raw material and ammonia to be contained are introduced into a pressure vessel, the inside of the pressure vessel is set to a desired pressure and temperature, and the treatment is performed for a desired time.
  • the ammonia may be in a liquid phase, a gas phase, or a supercritical state.
  • liquid ammonia or supercritical state Ammonia is suitable.
  • the conditions for the treatment with ammonia are not particularly limited and may be appropriately selected depending on the intended purpose.
  • the temperature is preferably ⁇ 35 ° C. to 140 ° C. and the pressure is preferably 0 MPa to 12.5 MPa.
  • the organic amine to be used is not particularly limited and can be appropriately selected according to the purpose.
  • ethylene diane examples thereof include monomethylamine and monoethylamine. These may be used alone or in combination of two or more. Among these, ethylenediamine is preferable.
  • the conditions such as the treatment temperature and pressure for treating the biomass containing cellulose type I with the organic amine are not particularly limited and can be appropriately selected depending on the purpose. For example, the same conditions as the treatment with ammonia Etc.
  • ammonia as the treatment agent from the viewpoint of the efficiency of transformation from cellulose type I to cellulose III type I , ease of removal of the treatment agent after treatment, and the like.
  • the time for the treatment with the treatment agent containing at least one of ammonia and organic amine is not particularly limited, and is appropriately determined depending on the amount of biomass raw material containing the cellulose type I to be used, the treatment pressure, the treatment temperature, and the like. Although it can be selected, it is preferably 10 minutes to 10 hours, more preferably 30 minutes to 8 hours, and particularly preferably 30 minutes to 5 hours. When the treatment time is less than 10 minutes, the treatment becomes insufficient, and the sugar production efficiency may be lowered. When the treatment time is longer than 10 hours, the treatment efficiency may be saturated, resulting in inefficiency as a whole. Sometimes. On the other hand, when the treatment time is within the more preferable range, it is advantageous in that the biomass raw material can be treated efficiently and the sugar production efficiency is improved.
  • the amount of at least one of the ammonia and the organic amine used in the treatment with the treatment agent containing at least one of the ammonia and the organic amine is not particularly limited and may be appropriately selected depending on the purpose. 10 mg to 300 g is preferable, 100 mg to 150 g is more preferable, and 1 g to 50 g is particularly preferable with respect to 1 g of the biomass raw material containing cellulose type I.
  • the amount of at least one of ammonia and organic amine used is less than 10 mg relative to 1 g of biomass raw material containing cellulose type I, the treatment may be insufficient. Efficiency can be poor.
  • the amount used is within the particularly preferable range, it is advantageous in that the treatment time can be shortened and the amount of the treatment agent to be used can be reduced.
  • other compounds include carbon dioxide, nitrogen, ethylene, methane, ethane, propane, butane, pentane, hexane, toluene, benzene, phenol, dioxane, xylene, acetone, chloroform, carbon tetrachloride, ethanol, methanol, and propanol. And butanol. These may be used alone or in combination of two or more.
  • the “modified biomass raw material” in the present invention means a material obtained by treating a biomass raw material containing cellulose type I with a treating agent containing at least one of ammonia and an organic amine. Cellulose I contained in the biomass raw material It is preferable that at least a part of the mold is transformed into cellulose III type I.
  • a modified biomass raw material can be obtained by treatment with a treatment agent containing at least one of ammonia and organic amine.
  • a treatment agent containing at least one of ammonia and organic amine By the treatment, at least a part of cellulose type I contained in the biomass raw material can be transformed into cellulose III type I having a lower crystal density.
  • Cellulose III type I is advantageous in that the enzyme easily acts because of its low crystal density.
  • the biomass feedstock by the process, the cellulose I-type and hemicellulose contained in the biomass material, such as respectively cellulose III I type or hemicellulose derived oligosaccharides, changes to a more enzyme acts prone state It is possible to improve enzyme saccharification efficiency.
  • the above-mentioned treatment removes modification by ester bonds such as acetyl groups and feruloyl groups in the biomass raw material, and the enzyme easily acts on the remaining hemicellulose in the biomass raw material, thereby further improving the sugar production efficiency. It is guessed.
  • the modified biomass raw material preferably contains cellulose III type I , and the ratio is not limited, but the more cellulose III type I , the more preferable in terms of obtaining excellent enzymatic saccharification efficiency.
  • the modified biomass material includes, for example, cellulose type I (cellulose I ⁇ type, cellulose I ⁇ type) and other components such as hemicellulose and lignin. May be.
  • lignin is not contained or the content thereof is small.
  • the modified biomass raw material as a method for confirming that at least a part of the cellulose I type is converted to the cellulose III I type, there is no particular limitation, and can be appropriately selected according to the purpose, For example, analysis methods such as X-ray diffraction, FT-IR, solid-state NMR and the like can be mentioned.
  • the modified biomass raw material can be obtained by treating a biomass raw material containing cellulose type I, which is a natural cellulose, with a treatment agent containing at least one of ammonia and organic amine. It may be in a state of a complex of cellulose and at least one of ammonia and organic amine (hereinafter, sometimes referred to as “complex of cellulose / ammonia”) produced in the treatment step.
  • a complex of cellulose / ammonia is difficult to adjust pH during enzymatic saccharification, and has the property of returning to cellulose type I by the action of water. It is preferable to use the modified biomass raw material in a state where at least one of ammonia and organic amine is removed from the composite of cellulose and ammonia.
  • the said process (B) is a process of obtaining the raw material for enzyme saccharification by grind
  • the modified biomass raw material obtained in the step (A) is pulverized.
  • a method for pulverizing the modified biomass raw material is not particularly limited and may be appropriately selected depending on the intended purpose.
  • a pulverizer such as a flat mill, a planetary ball mill, a vibration ball mill, a bead mill, a jet mill or the like may be used. And the method of using them.
  • a flat mill is preferable as the pulverizer because a fine powdery raw material for enzymatic saccharification excellent in enzymatic saccharification efficiency can be obtained with relatively low energy.
  • the modified biomass raw material can be used as the raw material for enzymatic saccharification of the present invention. By pulverizing the modified biomass raw material, it is possible to further improve the enzymatic saccharification efficiency.
  • the conditions for the pulverization are not particularly limited and may be appropriately selected depending on the type of pulverizer, the type of biomass raw material, the average particle size of the pulverized product to be obtained, and the like.
  • a flat mill is used as the pulverizer, the pulverized biomass raw material is discharged from the mortar. Therefore, when further pulverization is performed, the discharged pulverized material is collected and supplied to the mortar again. May be repeated multiple times.
  • the number of times of pulverization is not particularly limited, and can be appropriately selected according to the type of pulverizer used, the time per pulverization, the energy applied, and the like.
  • the pulverization is advantageous in that a finer powdery raw material for enzyme saccharification with excellent enzymatic saccharification efficiency can be obtained each time the pulverization is repeated. 4 times or less is preferable from the standpoint of inefficiency as a whole.
  • enzymatic saccharification raw material refers to a material obtained by pulverizing the modified biomass raw material. By crushing the modified biomass raw material, it is possible to further improve the enzymatic saccharification efficiency.
  • the particle size of the enzyme saccharification raw material particles obtained by the pulverization is not particularly limited, and the preferred size varies depending on the type of biomass raw material to be used.
  • the average particle size is preferably 5 ⁇ m to 80 ⁇ m, more preferably 5 ⁇ m to 50 ⁇ m, and even more preferably 5 ⁇ m to 30 ⁇ m.
  • the average particle size of the enzyme saccharification raw material is less than 5 ⁇ m, it takes a lot of energy and time for pulverization in order to make the average particle size less than 5 ⁇ m, thus losing economic rationality. Moreover, when the said average particle diameter exceeds 80 micrometers, enzyme saccharification efficiency may not fully improve. On the other hand, when the average particle size of the enzyme saccharification raw material is within the further preferable range, it is advantageous in terms of balance between energy and time required for pulverization and enzyme saccharification efficiency.
  • a median diameter obtained by measurement by a laser diffraction confusion method is adopted as the average particle diameter of the enzyme saccharification raw material in the present invention.
  • the median diameter refers to a particle diameter in which the cumulative volume of particles having a particle diameter equal to or larger than the particle diameter is equal to particles having a particle diameter equal to or smaller than the particle diameter.
  • the raw material for enzymatic saccharification obtained by pulverization of the modified biomass may be used, for example, as it is in the enzymatic saccharification of step (C) described later, or after appropriately passing through other steps, step (C) described later. It may be subjected to enzymatic saccharification.
  • the step (C) is a step of obtaining a sugar by enzymatic saccharification of the raw material for enzymatic saccharification obtained in the step (B) using cellulase and hemicellulase.
  • cellulase and hemicellulase are used in combination, which is advantageous in that the sugar production efficiency can be significantly improved.
  • “combination” refers to causing cellulase and hemicellulase to simultaneously act on the enzyme saccharification raw material.
  • the method for performing the enzymatic saccharification is not particularly limited as long as cellulase and hemicellulase are used in combination, and can be appropriately selected according to the purpose.
  • Cellulase is a general term for enzymes that hydrolyze the glycosidic bonds of ⁇ -1,4-glycans of cellulose.
  • the hemicellulase is a generic term for enzymes that hydrolyze the glycosidic bond of hemicellulose.
  • the hemicellulase is not particularly limited and may be appropriately selected depending on the intended purpose.
  • xylanase EC 3.2.1.8 or EC 3.2.1.136
  • mannanase EC 3. 2.1.78
  • ⁇ -xylosidase EC 3.2.1.37
  • ⁇ -mannosidase EC 3.2.1.25
  • ⁇ -glucuronidase EC 3.2.1.139
  • ⁇ -Arabinofuranosidase EC 3.2.1.55
  • ⁇ -galactosidase EC 3.2.1.22
  • acetyl xylan esterase EC 3.1.1.72
  • feruloyl esterase EC 3.1.73
  • etc. and may include ⁇ -glucanase (EC 3.2.1.6), pectinase (EC 3.2.1.15), and the like.
  • hemicellulase suitably according to the main component of a biomass raw material, etc.
  • the method for obtaining cellulase and / or hemicellulase is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include methods obtained from animals, plants, microorganisms and the like. Commercially available cellulase preparations and hemicellulase preparations may also be used.
  • Trichoderma Trichoderma
  • Acremonium genus Acremonium genus (Acremonium genus (Aspermonium) genus
  • Aspergillus Aspergillus
  • genus Phanerochaete genus Trametes, genus Humicola, genus Bacillus, genus Aureobasidium, genus Thermomyces, etc.
  • the total amount of cellulase and hemicellulase used in the enzyme saccharification is not particularly limited and may be appropriately selected depending on the intended purpose, but is 0.001 mg to 100 mg per 1 g of the enzyme saccharification raw material. Preferably, 0.01 mg to 10 mg is more preferable, and 0.1 mg to 1 mg is still more preferable. If the amount of the enzyme used is less than 0.001 mg with respect to 1 g of the enzyme saccharification raw material, enzyme saccharification may be insufficient, and if it exceeds 100 mg, saccharification inhibition may occur. On the other hand, when the amount of the enzyme used is within the further preferable range, it is advantageous in that the amount of sugar obtained is larger than the amount of enzyme added.
  • the amount of each of cellulase and hemicellulase used in the enzymatic saccharification is not particularly limited and may be appropriately selected according to the purpose.
  • the mass ratio of hemicellulase to cellulase is , 0.001 to 0.5 is preferable, and 0.01 to 0.2 is more preferable. If the mass ratio is less than 0.001, a sufficient amount of glucose may not be obtained, and if it exceeds 0.5, the amount of glucose obtained does not increase even if the amount of hemicellulase added is increased. Therefore, it is disadvantageous in terms of cost.
  • the mass ratio is within the preferable range, it is advantageous in that a large amount of sugar is obtained with respect to the amount of enzyme added.
  • the enzyme used in the enzymatic saccharification is not particularly limited as long as cellulase and hemicellulase are used.
  • other enzymes such as cellobiase (EC 3.2.1.21) may be used in combination.
  • the amount used is not particularly limited and may be appropriately selected depending on the intended purpose.
  • the mass ratio of the other enzyme to cellulase (others) is preferably 0.01 to 2, more preferably 0.1 to 1.0.
  • the temperature for the enzymatic saccharification is not particularly limited and may be appropriately selected depending on the intended purpose. For example, 10 ° C. to 70 ° C. is preferable, 20 ° C. to 60 ° C. is more preferable, and 30 ° C. to 50 ° C. More preferred is ° C. If the temperature is less than 10 ° C, enzyme saccharification may not proceed sufficiently, and if it exceeds 70 ° C, the enzyme may be deactivated. On the other hand, when the temperature is within the more preferable range, it is advantageous in that a large amount of sugar is obtained relative to the amount of enzyme added.
  • the time for performing the enzymatic saccharification is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably 1 hour to 96 hours, and more preferably 2 hours to 72 hours. If the time is less than 1 hour, enzyme saccharification may not proceed sufficiently, and if it exceeds 96 hours, the enzyme may be deactivated, which is not preferable from the viewpoint of sugar production efficiency. On the other hand, when the time is within the further preferable range, it is advantageous in that a large amount of sugar is obtained relative to the amount of enzyme added.
  • the pH for the enzymatic saccharification is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably 3.0 to 8.0, more preferably 3.5 to 7.0, and 4. 0 to 6.0 is more preferable.
  • the pH is less than 3.0 or exceeds 8.0, the enzyme may be deactivated.
  • the pH is within the more preferable range, it is advantageous in that the amount of sugar obtained is larger than the amount of enzyme added.
  • the cellulose I-type derived sugars, molasses, including sugar-derived hemicellulose be able to.
  • the sugar derived from cellulose III type I and the sugar derived from cellulose type I are not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include glucose.
  • the sugar derived from hemicellulose is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include pentoses such as xylose and arabinose, and hexoses such as glucose, galactose and mannose. It is done.
  • the sugar solution may be used, for example, as it is in the production method of the ethanol of the present invention or the production method of lactic acid of the present invention as it is, or after the other steps described later, You may use for the manufacturing method of the lactic acid of this invention.
  • the removal step is performed after the treatment with at least one of the ammonia and the organic amine, and then the cellulose / ammonia complex.
  • a removal step of removing at least one of ammonia and organic amine is not particularly limited, and can be appropriately selected depending on the purpose. For example, the method obtained after treatment with at least one of ammonia and organic amine is used.
  • Examples include a method of washing a modified biomass material containing a complex such as cellulose / ammonia with methanol, ethanol, acetone or the like, a method of drying under reduced pressure, a method of drying at a temperature higher than the boiling point of the treating agent, and the like.
  • a method of washing a modified biomass material containing a complex such as cellulose / ammonia with methanol, ethanol, acetone or the like a method of drying under reduced pressure, a method of drying at a temperature higher than the boiling point of the treating agent, and the like.
  • ammonia is used as the treating agent, the removal method is not using an organic solvent, and is excellent in safety.
  • the boiling point of ammonia for example, from room temperature to 50 ° C.
  • the method of drying is preferred.
  • the first pH adjustment step is a step of adjusting the enzyme saccharification raw material in the step (B) to a pH suitable for the enzyme saccharification.
  • the second pH adjustment step is a step of adjusting the sugar solution in the step (C) to a pH suitable for each fermentation step in the ethanol production method and lactic acid production method described later.
  • the reagent used for adjusting the pH is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include acids such as hydrochloric acid, sulfuric acid and formic acid, and alkalis such as sodium hydroxide and potassium hydroxide. .
  • the sugar obtained by the sugar production method is used to produce ethanol from biomass raw materials for the purpose of producing environmentally friendly fuel, to produce lactic acid using biomass raw materials that cannot be used for food, It can be suitably used for the production of degradable plastics.
  • the ethanol production method of the present invention includes a step (fermentation step) of fermenting the sugar obtained by the sugar production method of the present invention to obtain ethanol, and further includes other steps as necessary.
  • the method for fermenting the sugar is not particularly limited and may be appropriately selected according to the purpose.However, an alcohol-fermenting microorganism such as yeast is added to the solution containing the sugar, A method of performing alcoholic fermentation is preferred.
  • the yeast is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include Saccharomyces yeasts.
  • the yeast may be natural yeast or genetically modified yeast.
  • ethanol-fermenting microorganism examples include Saccharomyces cerevisiae, Kluyveromyces fragilis, Kluyveromyces lactis (K. lactis), Kluyveromyces marxianus Recombinant yeasts such as K. marxianus, Pichia stipitis, P. pastoris, Pachisolen tannofilus, Candida glabrata, or Candida globata Mobilis (Zymomonas mobilis), Simonova Tha Palme (Zymobacter palmae), Clostridium thermocellum (Clostridium thermocellum), and the like Clostridium Rujungudari (C.ljungdahlii) bacteria or these recombinants such. These may be used alone or in combination of two or more.
  • the amount of yeast used, fermentation temperature, pH, fermentation time and other conditions are not particularly limited, and are appropriately selected according to the amount of sugar to be used for alcohol fermentation, the type of yeast to be used, etc. can do.
  • Ethanol obtained by the ethanol production method can be suitably used as, for example, fuel ethanol, industrial ethanol, and the like. Since the ethanol can be obtained from a biomass raw material, it can be reproduced as long as the biomass raw material can be produced, and the plant absorbs carbon dioxide in the atmosphere at the time of cultivation. Even if carbon dioxide is generated by combustion, it does not increase the carbon dioxide concentration in the atmosphere. Therefore, it can be said that ethanol is a desirable energy source for preventing global warming. In recent years, such ethanol is particularly expected to be mixed with gasoline and used as an environmentally friendly automobile fuel.
  • An alcohol other than ethanol is produced by fermenting the sugar obtained by the sugar production method of the present invention using a microorganism that produces the desired alcohol, instead of the yeast that produces ethanol.
  • a microorganism that produces the desired alcohol instead of the yeast that produces ethanol.
  • the method for producing lactic acid according to the present invention includes a step of fermenting the saccharide obtained by the method for producing saccharide according to the present invention to obtain lactic acid (fermentation step), and further includes other steps as necessary.
  • the method for fermenting the sugar is not particularly limited and can be appropriately selected depending on the purpose, but a lactic acid-fermenting microorganism such as a lactic acid bacterium is added to the solution containing the sugar, A method of performing lactic acid fermentation is preferred.
  • the lactic acid bacterium is not particularly limited and may be appropriately selected depending on the intended purpose. Streptococcus thermophilus) and Lactobacillus bulgaricus. These may be used alone or in combination of two or more.
  • the lactic acid bacterium may be a natural lactic acid bacterium or a genetically modified lactic acid bacterium.
  • the amount of lactic acid bacteria used, fermentation temperature, pH, fermentation time, and other conditions are not particularly limited, and are appropriately selected according to the amount of saccharide used for lactic acid fermentation, the type of lactic acid bacteria used, and the like. can do.
  • the lactic acid obtained by the lactic acid production method can be suitably used for producing polylactic acid by chemical polymerization, for example.
  • lactic acid which is often produced from starch such as corn, from biomass raw materials containing cellulose that cannot be used for food.
  • the method for producing lactic acid such cellulose can be produced. It is possible to efficiently produce polylactic acid from a biomass raw material containing
  • the saccharide obtained by the sugar production method of the present invention is fermented using microorganisms that produce the desired organic acid in place of the lactic acid bacteria, so that organic acids other than lactic acid, such as citric acid, succinate Acid, malic acid, oxalic acid and the like can also be produced.
  • organic acids other than lactic acid such as citric acid, succinate Acid, malic acid, oxalic acid and the like can also be produced.
  • Example 1 Biomass raw material> Eucalyptus was used as a biomass raw material containing cellulose type I.
  • the prepared eucalyptus was coarsely ground using a Willet mill and the target average particle size was 200 ⁇ m.
  • ⁇ Ammonia treatment> The coarsely crushed eucalyptus was subjected to treatment with supercritical ammonia by the following operation.
  • a 4 g sample of coarsely crushed eucalyptus dried in an oven at 60 ° C. for 24 hours is transferred to a portable reactor TVS-N2 type having an internal volume of 120 mL (manufactured by Pressure Glass Industrial Co., Ltd .: hereinafter simply referred to as “container”).
  • the mixture was sealed and ammonia was allowed to flow in at a pressure of 0.5 MPa for 30 minutes while cooling the container to ⁇ 13 ° C. with a cooling device.
  • a PC-V type heater manufactured by Pressure Glass Industrial Co., Ltd.
  • a PC-V type heater manufactured by Pressure Glass Industrial Co., Ltd.
  • the pressure in the container was 11 MPa or more at which ammonia became a supercritical state.
  • the inside of the container was brought to atmospheric pressure to remove ammonia, the temperature was cooled to room temperature, and a solid sample in the container was collected. The ammonia was sufficiently evaporated overnight without sealing the sample to obtain an ammonia-treated product 1.
  • Enzymatic saccharification reaction solutions 1 to 8 were each shaken in a constant temperature room at 37 ° C. using a rotary shaker (15 rev / min) for 24 hours to perform enzymatic saccharification reaction.
  • the solution after the reaction is centrifuged, and the glucose concentration in the obtained supernatant is determined using the GOD-POD method (glucose oxidase-peroxidase method) as a glucose kit (trade name: Glucose CII Test Wako, manufactured by Wako Pure Chemical Industries, Ltd.). )
  • GOD-POD method glucose oxidase-peroxidase method
  • Glucose CII Test Wako manufactured by Wako Pure Chemical Industries, Ltd.
  • Comparative Example 1-1 Except that a sample obtained by pulverizing the ammonia-treated product 1 of Example 1 was used, and the enzyme saccharification reaction solutions 1 to 8 in the enzyme saccharification reaction of Example 1 were replaced with the enzyme saccharification reaction solution 9 prepared by the following method. By the same operation as in Example 1, the amount of glucose (g) per kg of biomass raw material in Comparative Example 1-1 was calculated.
  • Enzymatic saccharification reaction solution 9 was prepared in the same manner as enzymatic saccharification reaction solutions 1 to 8, except that the saccharification reaction solution 1 to 8 prepared in Example 1 was not added with the Trichoderma fungus-derived xylanase preparation. That is, 10 mg of a precisely weighed sample was taken into a 1.5 mL internal volume microtube, and the concentration of the sample was 1% (wt / vol).
  • Aspergillus cellobiase preparation (trade name: Novozym (registered trademark) 188, manufactured by Novozymes) with each enzyme concentration of 0.01% (wt / vol), total 0.02% (wt / vol)
  • An enzyme saccharification reaction solution 9 was prepared using a 50 mM acetate buffer (pH 4.5) so that the enzyme concentration was.
  • Example 1-2 The coarsely crushed eucalyptus used in Example 1 was pulverized by the same operation as the operation of pulverizing the sample after ammonia treatment in Example 1 without being ammonia-treated. Next, in Example 1, an enzyme saccharification reaction was performed by the same operation as in Example 1 except that the sample pulverized without ammonia treatment was used instead of the sample pulverized ammonia-treated product 1, and compared. The amount of glucose (g) per kg of biomass raw material in Example 1-2 was calculated. The results are shown in FIG.
  • Comparative Example 1-3 A sample pulverized without ammonia treatment of Comparative Example 1-2 was used except that enzyme saccharification reaction liquid 9 was used instead of enzyme saccharification reaction liquids 1 to 8 in the enzymatic saccharification reaction of Comparative Example 1-2.
  • the amount of glucose (g) per kg of biomass raw material of Comparative Example 1-3 was calculated by the same operation as Comparative Example 1-2.
  • Example 2 In Example 1, instead of Eucalyptus used as a biomass raw material, except for using Ezokinu willow, in the same operation as in Example 1, coarse pulverization treatment, ammonia treatment, pulverization treatment, and enzymatic saccharification reaction, By the same operation as in Example 1, the amount of glucose (g) per kg of biomass raw material in Example 2 was calculated. The results are shown in FIG. The treated product treated with ammonia in Example 2 may be referred to as “ammonia treated product 2”.
  • Comparative Example 2-1 A sample obtained by pulverizing the ammonia-treated product 2 of Example 2 was used except that an enzyme saccharification reaction solution 9 was used in place of the enzyme saccharification reaction solutions 1 to 8 in the enzyme saccharification reaction of Example 2. By the same operation, the amount of glucose (g) per kg of biomass raw material of Comparative Example 2-1 was calculated.
  • Example 2 The coarsely pulverized Ezo noki willow used in Example 2 was pulverized by the same operation as that for pulverizing the sample after ammonia treatment in Example 2 without ammonia treatment. Next, in Example 2, an enzyme saccharification reaction was performed in the same manner as in Example 2 except that the sample pulverized without ammonia treatment was used instead of the sample obtained by pulverizing the ammonia-treated product 2. The amount of glucose (g) per kg of biomass raw material in Example 2-2 was calculated. The results are shown in FIG.
  • Comparative Example 2-3 A sample pulverized without ammonia treatment of Comparative Example 2-2 was used except that enzyme saccharification reaction solution 9 was used instead of enzyme saccharification reaction solutions 1 to 8 in the enzyme saccharification reaction of Comparative Example 2-2. By the same operation as Comparative Example 2-2, the amount of glucose (g) per kg of biomass raw material of Comparative Example 2-3 was calculated.
  • Example 3 In Example 1, in place of eucalyptus used as a biomass raw material, sugarcane bagasse was used, and the coarse pulverization treatment, ammonia treatment, pulverization treatment, and enzymatic saccharification reaction were performed in the same manner as in Example 1. The amount of glucose (g) per kg of biomass raw material of Example 3 was calculated by the same operation as Example 1. The results are shown in FIG. The treated product treated with ammonia in Example 3 may be referred to as “ammonia treated product 3”.
  • Comparative Example 3-1 A sample obtained by pulverizing the ammonia-treated product 3 of Example 3 was used except that the enzyme saccharification reaction solution 9 was used in place of the enzyme saccharification reaction solutions 1 to 8 in the enzyme saccharification reaction of Example 3. By the same operation, the amount of glucose (g) per kg of biomass raw material of Comparative Example 3-1 was calculated.
  • Example 3-2 The coarsely pulverized Ezo noki willow used in Example 3 was pulverized by the same operation as that for pulverizing the sample after ammonia treatment in Example 3 without ammonia treatment. Next, in Example 3, an enzyme saccharification reaction was performed in the same manner as in Example 3 except that the sample pulverized without ammonia treatment was used instead of the sample obtained by pulverizing the ammonia-treated product 3. The amount of glucose (g) per kg of biomass raw material in Example 3-2 was calculated. The results are shown in FIG.
  • Comparative Example 3-3 A sample pulverized without ammonia treatment of Comparative Example 3-2 was used except that enzyme saccharification reaction solution 9 was used instead of enzyme saccharification reaction solutions 1 to 8 in the enzyme saccharification reaction of Comparative Example 3-2.
  • the amount of glucose (g) per kg of biomass raw material in Comparative Example 3-3 was calculated by the same operation as in Comparative Example 3-2.
  • Example 4 In Example 1, in place of eucalyptus used as a biomass raw material, switch grind was used, and coarse pulverization treatment, ammonia treatment, pulverization treatment, and enzymatic saccharification reaction were performed in the same manner as in Example 1. The amount of glucose (g) per kg of biomass raw material in Example 4 was calculated by the same operation as in Example 1. The results are shown in FIG. In addition, the treated product treated with ammonia in Example 4 may be referred to as “ammonia treated product 4”.
  • Comparative Example 4-1 A sample obtained by pulverizing the ammonia-treated product 4 of Example 4 was used except that the enzyme saccharification reaction solution 9 was used in place of the enzyme saccharification reaction solutions 1 to 8 in the enzyme saccharification reaction of Example 4. By the same operation, the amount of glucose (g) per kg of biomass raw material of Comparative Example 4-1 was calculated.
  • Example 4 The coarsely pulverized switch glass used in Example 4 was pulverized by the same operation as that for pulverizing the sample after ammonia treatment in Example 4 without being ammonia-treated.
  • Example 4 an enzymatic saccharification reaction was performed by the same operation as in Example 4 except that the sample pulverized without ammonia treatment was used instead of the sample obtained by pulverizing the ammonia-treated product 4.
  • the amount of glucose (g) per kg of biomass raw material in Example 4-2 was calculated. The results are shown in FIG.
  • Comparative Example 4-3 A sample pulverized without ammonia treatment in Comparative Example 4-2 was used except that enzyme saccharification reaction solution 9 was used instead of enzyme saccharification reaction solutions 1 to 8 in the enzyme saccharification reaction of Comparative Example 4-2. By the same operation as in Comparative Example 4-2, the amount of glucose (g) per kg of biomass raw material in Comparative Example 4-3 was calculated.
  • amount of glucose when using the enzyme saccharification reaction solution 9 is shown in Table 2.
  • the amount of glucose per kg of biomass was “(a)”, and the enzymatic saccharification reaction solution 9 (cellulase alone ), The amount of glucose per kg of biomass is “(b)”, and the increase in glucose increased by the combined use of cellulase and hemicellulase compared to the case where enzymatic saccharification reaction was performed with cellulase alone
  • the amount was calculated from the following formula (1).
  • the sugar production method, ethanol production method, and lactic acid production method of the present invention can significantly improve sugar production efficiency, ethanol production efficiency, and lactic acid production efficiency. Therefore, the sugar production method, the ethanol production method, and the lactic acid production method of the present invention are, for example, the production of ethanol from biomass raw materials for the purpose of producing environmentally friendly fuel, which has been attracting attention in recent years, It can be suitably used for producing environmentally friendly biodegradable plastics.

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Abstract

La présente invention concerne un procédé de production de sucre impliquant (A) une étape de traitement d'un matériau de type biomasse contenant de la cellulose de type I au moyen d'un agent de traitement contenant de l'ammoniac et/ou des amines organiques en vue de l'obtention d'un matériau de type biomasse modifiée, (B) une étape de broyage du matériau de type biomasse modifiée en vue de l'obtention d'un matériau pour saccharification enzymatique et (C) une étape de saccharification enzymatique dudit matériau pour saccharification enzymatique faisant appel à de la cellulase/ hémicellulase en vue de l'obtention d'un sucre.
PCT/JP2011/052861 2010-02-12 2011-02-10 Procédé de production de sucre, procédé de production d'éthanol et procédé de production d'acide lactique WO2011099558A1 (fr)

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WO2007130337A1 (fr) 2006-05-01 2007-11-15 Michigan State University Procédé de traitement d'une biomasse lignocellulosique
US10457810B2 (en) 2009-08-24 2019-10-29 Board Of Trustees Of Michigan State University Densified biomass products containing pretreated biomass fibers
WO2011028543A2 (fr) 2009-08-24 2011-03-10 Board Of Trustees Of Michigan State University Produits de biomasse densifiée et prétraitée, et procédés de fabrication et d'utilisation
US8945245B2 (en) 2009-08-24 2015-02-03 The Michigan Biotechnology Institute Methods of hydrolyzing pretreated densified biomass particulates and systems related thereto
CA2797193C (fr) 2010-04-19 2015-12-15 Board Of Trustees Of Michigan State University Biomasse lignocellulosique digestible, produits d'extraction du bois et procedes de production associes
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US10202660B2 (en) 2012-03-02 2019-02-12 Board Of Trustees Of Michigan State University Methods for increasing sugar yield with size-adjusted lignocellulosic biomass particles
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