CN115023507A - Method for producing sugar solution - Google Patents
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- CN115023507A CN115023507A CN202180011274.9A CN202180011274A CN115023507A CN 115023507 A CN115023507 A CN 115023507A CN 202180011274 A CN202180011274 A CN 202180011274A CN 115023507 A CN115023507 A CN 115023507A
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- 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
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- C13K1/00—Glucose; Glucose-containing syrups
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Abstract
A sugar solution in which fermentation inhibitors such as organic acids and aromatic compounds are reduced can be produced by a method for producing a sugar solution comprising the following steps (1) to (4). Step (1): a step (2) of pulverizing the cellulose-containing biomass so that the weight ratio of the cellulose-containing biomass that does not pass through a sieve having 1mm mesh is 50% or less by dry weight: a step of obtaining a pretreated biomass by mixing the pulverized biomass obtained in the step (1) with an alkaline aqueous medium, wherein the step (3): a step of adding water to the pretreated biomass obtained in the step (2) and performing solid-liquid separation to obtain a cellulose-containing solid component, wherein the step (4): and (3) hydrolyzing the cellulose solid obtained in step (3) to obtain a sugar solution.
Description
Technical Field
The present invention relates to a method for producing a sugar solution from a cellulose-containing biomass, the sugar solution being usable as a fermentation feedstock or the like.
Background
Fermentation processes for producing chemicals using sugars as raw materials are used for producing various industrial raw materials. As the sugar to be used as the fermentation raw material, substances derived from edible raw materials such as sugar cane, starch, and sugar beet are currently used industrially, but from the ethical side of the price explosion of edible raw materials due to the future increase in the world population or competition with eating, a process for producing a sugar solution more efficiently from cellulose-containing biomass, which is a renewable non-edible resource, or a process for efficiently converting the obtained sugar solution into an industrial raw material using the sugar solution as a fermentation raw material is a problem in the future.
Sugars in cellulose-containing biomass feedstocks are embedded in cell walls that form complex structures. Therefore, in order to allow the enzyme to function efficiently, it is preferable to subject the biomass material to alkali treatment before the enzymatic hydrolysis.
For example, it is disclosed that a sugar solution contains a sugar at a high purity by applying a specific pretreatment of bringing a cellulose-containing material into contact with an aqueous alkali solution and recycling an alkaline filtrate to a cellulose-containing biomass in order to increase the enzymatic hydrolysis rate of cellulose (patent document 1).
Further, there is disclosed a method for producing a raw material for enzymatic saccharification, which comprises treating a plant biomass raw material containing cellulose with a treating agent containing ammonia, immersing the treated raw material in water at 40 to 100 ℃ to elute polysaccharides in the water, and subjecting the resultant to the enzymatic saccharification step. According to the present invention, enzymatic saccharification can be efficiently performed, and therefore, the present invention can be used for a sugar production method with improved sugar production efficiency (patent document 2).
However, there is a quality problem that fermentation inhibitors such as organic acids and aromatic compounds generated during the alkali treatment are mixed into the sugar solution, and there is still a demand for a method for producing a cellulose sugar solution with reduced fermentation inhibitors.
Documents of the prior art
Patent document
Patent document 1: WO2017/170552
Patent document 2: WO2012/096236
Disclosure of Invention
Problems to be solved by the invention
The purpose of the present invention is to provide a method for reducing fermentation-inhibiting substances such as organic acids and aromatic compounds produced by alkali treatment of cellulose-containing biomass when producing a sugar solution from the cellulose-containing biomass.
Means for solving the problems
The present inventors have now found that a sugar solution with reduced fermentation inhibitor can be efficiently obtained if cellulose-containing biomass is pulverized to a specific pulverization degree, water is added by bringing the cellulose-containing biomass into contact with an alkaline aqueous medium, the pretreated biomass to which water has been added is subjected to solid-liquid separation, and the pretreated biomass is hydrolyzed.
The present invention is based on the above knowledge and is constituted by the following [1] to [10 ].
[1] A method for producing a sugar solution, which uses a cellulose-containing biomass as a raw material and comprises the following steps:
step (1): a step of pulverizing the cellulose-containing biomass so that the weight ratio of the cellulose-containing biomass that does not pass through a sieve having a mesh opening of 1mm is 50% or less by dry weight;
step (2): a step of bringing the pulverized biomass obtained in the step (1) into contact with an alkaline aqueous medium to obtain a pretreated biomass;
step (3): adding water to the pretreated biomass obtained in step (2) and performing solid-liquid separation to obtain a cellulose-containing solid component; and
step (4): and (3) hydrolyzing the cellulose-containing solid component obtained in step (3) to obtain a sugar solution.
[2] The method of producing a sugar solution according to [1], wherein the step (2) is a step of introducing an alkaline medium into the pulverized biomass obtained in the step (1) to obtain a pretreated biomass.
[3] The method of producing a sugar solution according to [1] or [2], wherein the step (2) is a step of supplying the pulverized biomass obtained in the step (1) and an alkaline medium to a filter, and allowing the pulverized biomass and the alkaline medium to pass through the filter.
[4] The method for producing a sugar solution according to any one of [1] to [3], wherein the alkaline aqueous medium in the step (2) is an aqueous medium containing sodium hydroxide and/or potassium hydroxide.
[5] The method for producing a sugar solution according to any one of [1] to [4], wherein the solid-liquid separation in the step (3) is squeezing.
[6] The method for producing a sugar solution according to any one of [1] to [5], wherein the cellulose-containing biomass is bagasse.
[7] The method for producing a sugar solution according to any one of [1] to [6], further comprising the following step (5): the sugar solution obtained in step (4) is filtered by a nanofiltration membrane or a reverse osmosis membrane, the sugar solution is recovered as a non-permeate, and the permeate is reused as the water added to the pretreated biomass in step (3).
[8] A cellulose-containing solid component comprising a pulverized cellulose-containing biomass and an alkaline aqueous medium, wherein the pulverized cellulose-containing biomass has a weight ratio of not passing through a sieve having a sieve opening of 1mm of 50% by weight or less in terms of dry weight, and the cellulose-containing solid component has a water content of 50% by weight or more and less than 70% by weight.
[9] The cellulose-containing solid component according to [8], wherein the cellulose-containing biomass is bagasse.
[10] A pulverized cellulose-containing biomass which does not pass through a sieve having a mesh size of 1mm, wherein the weight ratio of the pulverized cellulose-containing biomass to the dry weight is 40-50%.
ADVANTAGEOUS EFFECTS OF INVENTION
According to the present invention, a high-purity sugar solution with reduced fermentation-inhibiting substances derived from alkali treatment of cellulose-containing biomass can be obtained.
Drawings
FIG. 1 is a process flow chart showing an example of a method for producing a sugar solution of the present invention.
Detailed Description
The present invention is a method for producing a sugar solution using a cellulose-containing biomass as a raw material, comprising the steps of: step (1): a step of pulverizing the cellulose-containing biomass so that the percentage of the cellulose-containing biomass that does not pass through a sieve having a mesh opening of 1mm is 50% or less by dry weight; step (2): a step of bringing the pulverized biomass into contact with an alkaline aqueous medium to obtain a pretreated biomass; step (3): adding water to the pretreated biomass and performing solid-liquid separation to separate a cellulose-containing solid component; and a step (4): hydrolyzing the cellulose-containing solid component. Fig. 1 shows an example of a process flow chart of the present invention, but the process of the present invention is not limited to this.
It is known that the higher the degree of pulverization of the powder, the less dehydration is caused in the treatment of the drainage water in which the powder is slurried (see table 2 in j. soc. powder technol., Japan, 38, 177-183 (2001)). However, in the present invention, if water is added to pretreated biomass obtained by alkali-treating cellulose-containing biomass pulverized to a certain pulverization degree and solid-liquid separation is performed, the water content after solid-liquid separation is unexpectedly reduced as compared with the case where such pulverization is not performed, and thus a fermentation inhibitor derived from the pretreated biomass introduced at the time of hydrolysis in the later stage is reduced, and a high-purity sugar solution can be obtained.
The present invention will be described below according to the procedure.
Cellulose-containing biomass means a biomass containing at least cellulose. Examples of suitable cellulose-containing biomass include herbal biomass such as bagasse, switchgrass, elephant grass, festuca arundinacea, corn stalk, straw (rice straw, wheat straw), and oil palm empty fruit bunch, woody biomass such as trees, wood chips, and waste building materials, biomass derived from aquatic environment such as algae and seaweed, and grain husk biomass such as corn husk, wheat husk, soybean husk, and residue (cassava meal) obtained by extracting starch from cassava. Preferably herbaceous biomass such as bagasse, rice straw, and oil palm empty fruit bunch.
The water content of the cellulose-containing biomass is not particularly limited, and preferable ranges are, for example, 3% or more and 60% or less, 5% or more and 60% or less, 5% or more and 55% or less, and 5% or more and 50% or less. The water content of the cellulose-containing biomass was determined by the method detailed in the examples.
In the step (1), the cellulose-containing biomass is pulverized to a specific pulverization degree to obtain a pulverized biomass. The pulverizing means is not particularly limited, and can be performed by using a machine commonly used for coarse pulverization of various materials such as a ball mill, a vibration mill, a chopper mill, a hammer mill, a Vickers mill, and a jet mill. The mechanical pulverization may be either dry or wet, but is preferably dry pulverization.
As the pulverization degree of the pulverized biomass obtained in the step (1), the pulverization is performed until the weight ratio (dry weight%) of the pulverized biomass not passing through the sieve having a mesh size of 1mm becomes about 50% or less by dry weight, and the pulverization is performed until the weight ratio becomes preferably about 40% or more and about 50% or less, more preferably about 40% or more and about 45% or more and about 50% or less, whereby the water content after the unexpected solid-liquid separation is reduced as compared with the case of not performing such pulverization, whereby the fermentation inhibiting substance derived from the pretreated biomass introduced at the time of the later hydrolysis can be reduced. The degree of pulverization was evaluated by drying the pulverized biomass to a water content of 20% or less and then sieving the pulverized biomass through a sieve having a mesh opening of 1 mm. In the present specification, if the weight ratio (dry weight%) of the pulverized biomass that does not pass through the sieve having an opening of 1mm is low, the pulverization degree may be expressed as high. The method/conditions of the sieve are according to ISO 2591-1.
In the step (2), the pulverized biomass obtained in the step (1) is brought into contact with an alkaline aqueous medium to obtain a pretreated biomass. Here, the pretreated biomass refers to a biomass containing a solid component of the cellulose-containing biomass subjected to alkali treatment for efficiently hydrolyzing the cellulose-containing biomass in the subsequent step (4).
The method of bringing the pulverized biomass obtained in step (1) into contact with the alkaline aqueous medium is not particularly limited as long as it is a cellulose-containing biomass alkali treatment method well known to those skilled in the art, and the alkali treatment method described in WO2017/170552 is preferably used. The method is a method for obtaining pretreated biomass and an alkali treatment solution rich in coumaric acid and ferulic acid by introducing an alkaline aqueous medium into a cellulose-containing biomass.
As a method for passing the alkaline aqueous medium through the pulverized biomass, there is a method in which the pulverized biomass and the alkaline aqueous medium are supplied to a filter and the pulverized biomass and the alkaline medium are passed through the filter according to the method described in WO 2017/170552. The pulverized biomass and the alkaline aqueous medium may be supplied to the filter by being mixed in advance, or may be supplied to the filter separately.
The pH of the alkaline filtrate may be in the same range as that of the alkaline aqueous medium, and a preferable range of pH is, for example, 7 or more and 12 or less, 8 or more and 12 or less, a more preferable range of pH is 9 or more and 12 or less, and a further preferable range of pH is 10 or more and 12 or less. The pH of the alkaline filtrate has a tendency to decrease as the reaction proceeds. This is because if the alkali reaction proceeds, the soluble lignin component functions as a neutralizing agent, and the progress of the reaction can be estimated from the degree of the decrease. In particular, the pH range of the pulverized biomass after (after) the pulverized biomass is brought into contact with the alkaline aqueous medium can be appropriately adjusted by the initial alkali concentration or the like, and is preferably 7 or more and 12.5 or less, 8 or more and 12.5 or less, more preferably 8 or more and 12 or less, and still more preferably 8 or more and 11 or less. Determining whether the pH of the alkaline filtrate is within the above range is an effective means in evaluating whether the reaction has proceeded to a level sufficient for the subsequent hydrolysis step.
The alkaline aqueous medium in contact with the pulverized biomass is preferably kept at a substantially constant temperature. When the filter includes a filtering unit for passing the alkaline aqueous medium through the pulverized biomass, it is also preferable that the alkaline filtrate filtered from the filtering unit is kept at a substantially constant temperature. The temperature of the alkaline aqueous medium or the alkaline filtrate can be maintained by providing the filter with a known heat retention device or heating device.
The number of times of repeating the contact of the filtrate from the filtration part after the contact with the alkaline aqueous medium and the contact with the pulverized biomass again is not particularly limited, and is, for example, at least 2 times or more, 2 times or more and 20000 or less, 2 times or more and 10000 or less, 2 times or more and 1000 or less, 3 times or more and 10000 or less, 3 times or more and 1000 or less, or 3 times or more and 100 or less.
The basic aqueous medium includes an alkaline aqueous solution such as ammonia, aqueous ammonia, or an aqueous medium containing a hydroxide, and is preferably an aqueous medium containing sodium hydroxide and/or potassium hydroxide, more preferably an aqueous medium containing sodium hydroxide, and still more preferably an aqueous solution of sodium hydroxide and/or potassium hydroxide.
The alkali concentration of the alkaline aqueous medium can be calculated from the content of an alkaline substance (an alkaline solid component such as hydroxide) in the alkaline-containing medium. The upper limit of the alkali concentration of the basic aqueous medium is not particularly limited, but is preferably about 3, 2, 1.5, 1, 0.7, 0.6, 0.5, 0.4, 0.3 or 0.2% by weight, and the lower limit is preferably about 0.05, 0.1, 0.2, 0.3, 0.4 or 0.5% by weight. Further, the alkali concentration is preferably in the range of, for example, 0.05 wt% or more and 0.3 wt% or less, 0.1 wt% or more and 3 wt% or less, 0.1 wt% or more and 2 wt% or less, more preferably in the range of 0.1 wt% or more and 2 wt% or less, 0.25 wt% or more and 1.5 wt% or less, and still more preferably in the range of 0.25 wt% or more and 1.0 wt% or less.
The lower limit of the pH of the basic aqueous medium is not particularly limited as long as it is basic, but is 7 or more, preferably 8 or more, more preferably 9 or more, and still more preferably 10 or more. The upper limit of the pH is not particularly limited as long as it is less than 14, but it can be set at pH13.5 or less from the viewpoint of reducing the amount of the base used. Further, a preferable range of pH is, for example, 7 or more and 13.5 or less, 8 or more and 13.5 or less, a more preferable range of pH is 9 or more and 13.5 or less, and a further more preferable range of pH is 10 or more and 13.5 or less.
The upper limit of the temperature of the basic aqueous medium is not particularly limited, but is preferably about 110, 100, 95, 90, 80, 75, or 70 ℃, and the lower limit is preferably about 35, 40, 50, 60, or 65 ℃. Further, the temperature of the basic aqueous medium is preferably in a range of, for example, 35 ℃ or more and 100 ℃ or less, 40 ℃ or more and 100 ℃ or less, 50 ℃ or more and 100 ℃ or less, 60 ℃ or more and 100 ℃ or less, 65 ℃ or more and 100 ℃ or less, 80 ℃ or more and 100 ℃ or less, more preferably in a range of 60 ℃ or more and 100 ℃ or less, 65 ℃ or more and 100 ℃ or less, 80 ℃ or more and 100 ℃ or less, and still more preferably in a range of 65 ℃ or more and 100 ℃ or more.
The weight ratio of the alkaline aqueous medium to the pulverized biomass (dry weight) is not particularly limited, and a preferable range is, for example, 100: 1-2: 1. 90: 1-3: 1. 50: 1-5: 1. 30: 1-5: 1. 25: 1-7: 1. 25: 1-5: 1. 20: 1-5: 1.
the ratio of the alkali-containing aqueous medium to the pulverized biomass (dry weight) may be set using, as an index, the amount of alkali used (also referred to as the amount of alkali reaction) calculated by the method for calculating the amount of alkali reaction described in the examples. The preferable range of the amount of the base used is, for example, 20mg/g to 300mg/g, 30mg/g to 200mg/g, 40mg/g to 200mg/g, 45mg/g to 180mg/g, 45mg/g to 150mg/g, 50mg/g to 120mg/g, 60mg/g to 120mg/g, and more preferably 45mg/g to 150mg/g, 50mg/g to 120mg/g, and 60mg/g to 120 mg/g.
The time for bringing the pulverized biomass into contact with the alkaline aqueous medium is not particularly limited, and the contact time is, for example, 20 minutes or more and 72 hours or less, 20 minutes or more and 48 hours or less, 20 minutes or more and 24 hours or less, 30 minutes or more and 48 hours or less, 30 minutes or more and 24 hours or less, 30 minutes or more and 12 hours or less, 30 minutes or more and 6 hours or less, or 30 minutes or more and 3 hours or less.
The pretreated biomass obtained by bringing the pulverized biomass into contact with an alkaline aqueous medium is preferably separated/recovered by a method well known to those skilled in the art, and is supplied to the subsequent step (3). When the filter is used when the pulverized biomass is brought into contact with the alkaline aqueous medium, the solid matter remaining in the filter can be recovered as the pretreated biomass.
The water content of the pretreated biomass supplied to the step (3) is not particularly limited, and is preferably in a range of, for example, 50 wt% or more and 99 wt% or less, 60 wt% or more and 99 wt% or less, 70 wt% or more and 99 wt% or less, 80 wt% or more and 99 wt% or less, or 80 wt% or more and 95 wt% or less, more preferably 80 wt% or more and 99 wt% or less, and still more preferably 80 wt% or more and 95 wt% or less. The water content of the pretreated biomass was measured by the method described in detail in the examples.
In the step (3), water is added to the pretreated biomass to separate the fermentation-inhibiting substance from the pretreated biomass, and solid-liquid separation is performed. By this step, it is possible to reduce the fermentation inhibiting substance which is generated by the alkali treatment of the cellulose-containing biomass contained in the pretreatment biomass and which is considered to be derived from lignin in the cellulose-containing biomass. Specific examples of the fermentation-inhibiting substance include organic acids such as formic acid, acetic acid, citric acid, and lactic acid, ferulic acid, coumaric acid, vanillin, vanillic acid, acetyl vanillin, 4-hydroxybenzoic acid, syringic acid, and aromatic compounds such as gallic acid, and furan-based compounds such as HMF and furfural.
The amount of water to be added to the pretreated biomass is not particularly limited, and is preferably in the range of 1: 1-1: 100. 1: 1-1: 50. 1: 1-1: 30. 1: 1-1: 20. 1: 1-1: 10. 1: 1-1: 5. 1: 1-1: 3.
the pH of the water added to the pretreated biomass is not particularly limited, and is preferably 3 to 9, 3 to 8, 3 to 7, 4 to 9, 4 to 8, 4 to 7, 5 to 9, 5 to 8, and 5 to 7.
The temperature of the water added to the pretreated biomass is not particularly limited, and preferable ranges are, for example, 10 to 60 ℃, 10 to 50 ℃, 10 to 40 ℃, 20 to 60 ℃, 20 to 50 ℃, 20 to 40 ℃, 30 to 60 ℃, 30 to 50 ℃ and 30 to 40 ℃.
As a method of adding the water-pretreated biomass, the pretreated biomass may be mixed with the added water in the reaction tank, or the reaction tank may be provided with a stirrer. The reaction tank is not limited in shape as long as the pretreated biomass can be brought into contact with the added water. The pretreated biomass may be conveyed to the reaction tank by a conveyor belt, natural dropping, or the like. In addition, as long as the pretreated biomass can be brought into contact with water, water may be added during conveyance by a conveyor or the like, even in a reaction tank.
As the apparatus used for solid-liquid separation, centrifugal separation, pressing, or the like can be applied, and pressing is preferable. The press may be a screw press, a belt press, a filter press, or the like, but a screw press is preferred.
From the viewpoint of reducing the fermentation inhibiting substances derived from the pretreated biomass, the water content of the cellulose-containing solid component after the solid-liquid separation is preferably about 50 wt% or more and about 85 wt% or less, and more preferably about 50 wt% or more and less than 70 wt%. The water content of the cellulose-containing solid component was measured by the method for measuring the water content described in the examples.
The content of the alkaline aqueous medium in the cellulose-containing solid component is preferably 5 mg/g-biomass or more and 40 mg/g-biomass or less, more preferably 7 mg/g-biomass or more and 35 mg/g-biomass or less, and still more preferably 10 mg/g-biomass or more and 30 mg/g-biomass or less. By reducing the content of the alkaline aqueous medium in the cellulose-containing solid content to the above range, the fermentation inhibiting substance derived from the pretreated biomass introduced at the time of the latter hydrolysis can be reduced.
In step (4), the cellulose-containing solid component is hydrolyzed to obtain a sugar solution. In the hydrolysis step, a known hydrolysis method such as acid hydrolysis, alkali hydrolysis, or enzyme hydrolysis may be used, but it is preferable to subject the cellulose-containing solid component to hydrolysis treatment with an enzyme in an aqueous medium. The sugar solution obtained by such an enzymatic hydrolysis step can be obtained as an aqueous solution containing oligosaccharides and/or monosaccharides typified by glucose, xylose, arabinose, galactose, xylobiose, and cellobiose, for example.
The enzyme to be used is not particularly limited as long as it is a cellulose or hemicellulose hydrolase, and for example, a relatively inexpensive commercially available product, and as the cellulase enzyme agent, "Acremonium cellulase" (Meiji Seika ファルマ Co., Ltd.), enzyme "アクセルレース. デュエット" (ダニスコ. ジャパン Co., Ltd.), enzyme "Celluclast" 1.5L (ノボザイム Co., Ltd.) or the like, which is an enzyme derived from Acremonium, can be used. The hemicellulase enzyme may be "Optimash BG" (manufactured by ジェネンコア), or the like. The source of the enzyme is not particularly limited, but is more preferably an enzyme derived from a filamentous fungus. Enzymes derived from filamentous fungi abundantly include a cellulase, hemicellulase, β -glucosidase and other lytic enzymes derived from polysaccharides in cellulose-containing biomass, and are advantageous in hydrolysis reaction of biomass after alkali treatment.
The enzymes used may be used alone or in combination in consideration of the nature of the enzymatic agent, the composition of the desired product, and the like. The amount of the enzyme is not particularly limited and can be determined appropriately. The amount of such an enzyme may be, for example, 0.01g to 1g, preferably 0.001g to 0.1g, per 1g of the starting substrate. The temperature, pH and time of the enzymatic hydrolysis may be appropriately set depending on the nature, combination and the like of the enzyme. The temperature is, for example, 30 to 60 ℃ inclusive, and more preferably 35 to 50 ℃ inclusive. The pH is, for example, 3 or more and 8 or less, preferably 4 or more and 7 or less. The reaction time is, for example, 1 hour or more and 48 hours or less, preferably 6 hours or more and 24 hours or less.
The concentration of glucose, xylose or xylobiose in the sugar solution obtained in step (4) is not particularly limited, and can be appropriately set by adjusting the reaction conditions of the respective steps. Suitable glucose concentrations are, for example, about 5g/L to about 1000g/L, about 5g/L to about 700g/L, about 5g/L to about 550g/L, or about 10g/L to about 550 g/L. Further, the xylose concentration is preferably, for example, about 1g/L to about 100g/L, about 1g/L to about 50g/L, or about 1g/L to about 10 g/L. Further, the preferable concentration of xylobiose is, for example, about 1g/L to about 100g/L, about 1g/L to about 50g/L, about 1g/L to about 20g/L, or about 1g/L to about 15 g/L.
The sugar solution produced in step (4) can be used as it is for various industrial applications, but may be subjected to post-treatment as needed, specifically, treatment such as membrane treatment, centrifugal separation, concentration, and drying.
Preferable membrane treatment of the sugar solution produced in step (4) includes nanofiltration membrane treatment or reverse osmosis membrane treatment.
The nanofiltration membrane is also called a nanofilter (nanofiltration membrane, NF membrane), and is a membrane generally defined as a membrane that allows monovalent ions to pass through and blocks divalent ions. The membrane is considered to have a fine space of about several nanometers, and is used mainly for preventing fine particles, molecules, ions, salts, and the like in water.
The reverse osmosis membrane is also called RO membrane, and is a membrane generally defined as "a membrane having a desalination function by containing 1-valent ions", and is a membrane having an ultrafine pore of about several angstroms to several nanometers, and is mainly used for removing ionic components such as seawater desalination and ultrapure water production.
The nanofiltration membrane or reverse osmosis membrane treatment is a treatment in which the sugar solution produced in step (4) is filtered by a nanofiltration membrane or reverse osmosis membrane treatment, the dissolved sugar, in particular, monosaccharide such as glucose or xylose, oligosaccharide such as xylobiose or cellobiose is prevented or separated by filtration on the non-permeable side of the membrane, and the fermentation-inhibiting substance remaining in the sugar solution is allowed to permeate as a permeate, and can be carried out by the method described in WO 2010/067785.
The amount of fermentation-inhibiting substances in the sugar solution recovered from the non-permeable side of the nanofiltration membrane or reverse osmosis membrane is further reduced as compared with the amount before the membrane treatment, the fermentation performance can be improved as compared with the amount before the membrane treatment, and the permeate recovered from the permeable side of the nanofiltration membrane or reverse osmosis membrane can be used as the water added to the pretreated biomass in the step (3) (step (5)). This step can provide an unexpected effect that the amount of water used in all the steps can be reduced and the aromatic compound as the fermentation inhibitor can be further removed by the solid-liquid separation in the step (3).
Examples
The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples. Unless otherwise specified, the units and the measurement methods described in the present specification are based on Japanese Industrial Standards (JIS).
Various analysis methods in this example are as follows.
[ method of measuring sugar concentration ]
The concentration of sugars contained in the saccharified solutions obtained in the respective examples and comparative examples was determined by comparison with a standard under the following High Performance Liquid Chromatography (HPLC) conditions.
Column: luna NH 2 (manufactured by Phenomenex K.K.)
Mobile phase: ultrapure water; acetonitrile 25: 75
Flow rate: 0.6mL/min
Reaction solution: is composed of
The detection method comprises the following steps: RI (differential refractive index)
Temperature: at 30 ℃.
[ method for measuring the concentration of furan-based/aromatic-based Compound ]
The concentrations of furan-based compounds (HMF, furfural) and phenol-based compounds (vanillin, etc.) contained in the sugar solution were analyzed by HPLC under the conditions shown below, and the concentrations were quantified by comparison with a standard.
Column: synergi HidroRP 4.6 mm. times.250 mm (manufactured by Phenomenex)
Mobile phase: acetonitrile-0.1% H 3 PO 4 (flow rate 1.0mL/min) detection method: UV (283nm)
Temperature: at 40 deg.c.
[ method of measuring concentration of organic acid ]
The organic acids (acetic acid and formic acid) contained in the sugar solution were analyzed by HPLC under the conditions shown below, and the amounts were determined by comparison with standard substances.
Column: Shim-Pack SPR-H and Shim-Pack SCR101H (Shimadzu corporation) connected in series
Mobile phase: 5mM p-toluenesulfonic acid (flow 0.8mL/min)
Reaction solution: 5mM p-toluenesulfonic acid, 20mM Bis-Tris, 0.1mM EDTA-2 Na (flow rate 0.8mL/min)
The detection method comprises the following steps: electrical conductivity of
Temperature: at 45 ℃.
[ method for measuring Water content ]
The water contents of the cellulose-containing biomass, pretreated biomass, and cellulose-containing solid components used in the following experiments were measured. The water content (wt%; hereinafter, expressed in% only) was measured as a value obtained by subtracting the initial value from the stability value after evaporation by maintaining the sample at 120 ℃ using an infrared moisture meter ("FD-720", manufactured by ケット scientific research).
The water content of each cellulose-containing biomass measured by the present method is shown in table 1 for reference. Bagasse, rice straw, oil palm empty fruit bunches are classified as herbaceous biomass.
[ Table 1]
| Raw materials | Water content ratio |
| Bagasse | 50％ |
| Rice straw | 10％ |
| Oil palm empty fruit cluster | 15％ |
| Wood flour (fir) | 5％ |
[ method for calculating the amount of alkali reaction ]
The alkali reaction amount can be calculated, for example, by adding an aqueous solution b (g) of sodium hydroxide of y (%) to a cellulose-containing biomass raw material a (g) having a water content of x (%) and reacting the mixture, and calculating the alkali reaction amount (unit: mg/g-dry biomass) from the following formula 1.
The reaction amount of the base was y × b × 1000/{ (100-x) × a } · (formula 1).
Comparative reference example 1: relationship between bagasse pulverization degree and moisture content of cellulose-containing solid matter in hydrothermal treatment
In the step (1), bagasse having a water content of 50% was pulverized by a pulverizer (バリオニクス BRX-400, manufactured by Nara machinery Co., Ltd.). The pulverization conditions were that the pulverization was carried out while feeding the pulverized material at a rotation speed of 600rpm and a feed speed of 1000kg/hr with the mesh size of the chopper set to 30 mm. In order to measure the dry weight ratio, the pulverized biomass was dried to have a water content of 20%, passed through a sieve having a mesh size of 1mm under the conditions of ISO 2591-1, and the weight ratio of the pulverized biomass which did not pass was measured to be 45%.
In the step (2), the pulverized bagasse (with a water content of 50%) was subjected to hydrothermal treatment at 180 ℃ under high pressure for 10 minutes with a solid content of 5%. The resulting pretreated bagasse was filtered through a stainless steel strainer (opening ratio: 30%) having a mesh opening of 3mm, and the solid matter (pretreated biomass) remaining on the top surface of the strainer was pressed against the strainer surface by hand. The water content of the obtained pretreated biomass was 90%.
In step (3), water in an amount of 1.6 times the solid content (dry weight) was added to the pretreated biomass, and the pretreated biomass to which water was added was subjected to solid-liquid separation using a laboratory small screw press (HX 100, frequency 10Hz, manufactured by Fukuwa Industrial Co., Ltd.). The water content of the cellulose-containing solid component after the solid-liquid separation was measured, and as a result, the water content was 75%. The results are shown in table 2.
Comparative reference example 2: relationship between degree of pulverization of bagasse in hydrothermal treatment and water content of cellulose-containing solid matter
The pulverization was carried out under the same conditions as in comparative example 1 except that the mesh size of the chopper was 40 mm. In order to measure the dry weight ratio, the pulverized biomass was dried to a water content of 20%, passed through a sieve having an opening of 1mm under the conditions of ISO 2591-1, and the weight ratio of the pulverized biomass which did not pass through the sieve was measured to be 50%. The moisture content of the cellulose-containing solid component after solid-liquid separation was 73%. The results are shown in table 2.
Comparative reference example 3: relationship between degree of pulverization of bagasse in hydrothermal treatment and water content of cellulose-containing solid matter
The pulverization was carried out under the same conditions as in comparative example 1 except that the mesh opening size of the chopper was 50 mm. In addition, in order to determine the dry weight ratio, the pulverized biomass was dried to a water content of 20%, passed through a sieve having an opening of 1mm under the conditions of ISO 2591-1, and the weight ratio of the pulverized biomass which did not pass through the sieve was determined to be 55%. The water content of the cellulose-containing solid component after solid-liquid separation was 70%. The results are shown in table 2.
Comparative reference example 4: relationship between bagasse pulverization degree and moisture content of cellulose-containing solid component in alkali treatment
In the step (1), pulverization was carried out under the same pulverization conditions as in comparative example 3 except that the pulverization conditions were set such that the mesh size of the chopper was 50 mm. In order to measure the dry weight ratio, the pulverized biomass was dried to a water content of 20%, passed through a sieve having an opening of 1mm under the conditions of ISO 2591-1, and the weight ratio of the pulverized biomass which did not pass through the sieve was measured to be 55%.
In the step (2), 5.0kg of the obtained pulverized bagasse (having a water content of 50%) was charged into a multi-purpose extractor (manufactured by イズミフードマシナリ), 45kg of an aqueous sodium hydroxide solution having a predetermined concentration (initial temperature: 90 ℃ C., pH of about 13) was added from a spray ball in the upper part of a tank of the multi-purpose extractor, and a liquid (alkaline filtrate) obtained by filtration by its own weight from a filter net attached to the tank was added again from the spray ball, and the operation was repeated. Further, a heating mechanism is provided between the filter screen and the upper spray ball, and the reaction is carried out for a predetermined time while monitoring the temperature. In the reaction, the alkaline filtrate is adjusted in such a manner that it does not fall below 90 ℃. Further, the bagasse and the cellulose-containing solid matter are placed on the filter screen without using a stirring blade attached to the multi-function extractor, and the operation of shaping or slurrying with a stirring blade or the like is not performed. The alkaline filtrate was continuously circulated for a prescribed reaction time. The obtained sample was further filtered by a stainless steel strainer (opening ratio: 30%) having a mesh opening of 3mm, and the solid component (pretreated biomass) remaining on the top surface of the strainer was pressed against the strainer surface by hand. The water content of the obtained pretreated biomass was 90%.
In step (3), water in an amount of 1.6 times the solid content (dry weight) was added to the pretreated biomass, and the pretreated biomass was subjected to solid-liquid separation using a laboratory small screw press. The water content of the cellulose-containing solid component after the solid-liquid separation was measured, and as a result, the water content was 75%. The results are shown in table 2.
Reference example 1: relationship between degree of pulverization of bagasse in alkali treatment and water content of cellulose-containing solid component
The pulverization was carried out under the same conditions as in comparative reference example 4 except that the mesh opening size of the chopper was 40 mm. In order to measure the dry weight ratio, the pulverized biomass was dried to a water content of 20%, passed through a sieve having an opening of 1mm under the conditions of ISO 2591-1, and the weight ratio of the pulverized biomass which did not pass through the sieve was measured to be 50%. The moisture content of the cellulose-containing solid component after solid-liquid separation was 64%. The results are shown in table 2.
Reference example 2: relationship between degree of pulverization of bagasse in alkali treatment and water content of cellulose-containing solid component
The pulverization was carried out under the same conditions as in comparative reference example 4 except that the mesh size of the chopper was 30 mm. Further, in order to measure the dry weight ratio, the pulverized biomass was dried to have a water content of 20%, and passed through a sieve having an opening of 1mm under the conditions of ISO 2591-1, and the weight ratio was measured to be 45%. The moisture content of the cellulose-containing solid component after solid-liquid separation was 60%. The results are shown in table 2.
[ Table 2]
As can be seen from table 1, when the pretreatment is hydrothermal treatment, the percentage of pulverized biomass not passing through a sieve having a sieve opening of 1mm (dry weight%) is decreased (the pulverization degree is increased), and the cellulose-containing solid content moisture content (%) after solid-liquid separation is increased, which is a result of information well known to those skilled in the art that dehydration is more difficult as the pulverization degree of the powder is higher in the treatment of the wastewater in which the powder is slurried (see table 2 of j.soc.powder technology, Japan, 38, 177-183 (2001)). On the other hand, in the case where the pretreatment is the alkali treatment, as the proportion of the pulverized biomass not passing through the sieve having a mesh size of 1mm (dry weight%) decreases (the pulverization degree increases), contrary to the information well known to those skilled in the art, the moisture content (%) of the cellulose-containing solid component after the solid-liquid separation decreases, and it has been found that the possibility of the fermentation inhibitor derived from the pretreatment being mixed in the production of a sugar solution by the hydrolysis in the later stage can be reduced by increasing the pulverization degree of the pulverized biomass.
Comparative example 1: production example of sugar solution from bagasse (Steps (1) and (3) were not performed)
The same pulverization conditions and pretreatment conditions as in comparative reference example 4 were carried out to obtain a pretreated biomass. The resulting pretreated biomass was not subjected to the addition of water/solid-liquid separation in step (3), and pure water and 35% hydrochloric acid were added in step (4) so that the dry solids concentration became 5% and the pH became 5.0, to prepare a slurry containing cellulose-containing solids. To 500mL of the resulting slurry solution, 5mL of "アクセルレース & デュエット" as an enzyme manufactured by ダニスコ & ジャパン K.K., was added, and the reaction was carried out for 8 hours while maintaining the slurry temperature at 50 ℃ and constantly stirring. The sugar, acetic acid, and aromatic compound concentrations of the obtained reaction solution were measured, and the weight of acetic acid and coumaric acid was determined based on 100g of the obtained glucose, and the results are shown in table 3.
Comparative example 2: production example of sugar solution from bagasse (step (1) was not performed)
The same pulverization conditions, pretreatment conditions, and water-addition/solid-liquid separation conditions as in comparative reference example 4 were applied. The same hydrolysis operation as in comparative example 1 was performed as step (4) using the cellulose-containing solid component after the solid-liquid separation. The sugar, acetic acid, and aromatic compound concentrations of the obtained reaction solution were measured, and the weight of acetic acid and coumaric acid was determined based on 100g of the obtained glucose, and the results are shown in table 3.
Comparative example 3: production example of sugar solution from bagasse (step (3) is not performed)
The same operation as in comparative reference example 4 was carried out except that the crushing conditions were changed to 40mm mesh size in the chopper and solid-liquid separation was carried out without adding water to the pretreated biomass. The same hydrolysis operation as in comparative example 1 was performed as step (4) using the obtained cellulose-containing solid component after solid-liquid separation. The sugar, acetic acid, and aromatic compound concentrations of the obtained reaction solution were measured, and the weight of acetic acid and coumaric acid was determined based on 100g of the obtained glucose, and the results are shown in table 3.
Example 1: production example of sugar solution from bagasse (Steps (1) to (4) are carried out)
The same hydrolysis operation as in comparative example 1 was carried out as step (4) using the cellulose-containing solid component obtained in reference example 1 after the solid-liquid separation. The sugar, acetic acid, and aromatic compound concentrations of the obtained reaction solution were measured, and the weights of acetic acid and coumaric acid were determined based on 100g of the obtained glucose, and the results are shown in table 3.
Example 2: production example of sugar solution from bagasse (Steps (1) to (4) are carried out)
The same hydrolysis operation as in comparative example 1 was carried out as step (4) using the cellulose-containing solid component obtained in reference example 2 after the solid-liquid separation. The sugar, acetic acid, and aromatic compound concentrations of the obtained reaction solution were measured, and the weight of acetic acid and coumaric acid was determined based on 100g of the obtained glucose, and the results are shown in table 3.
Example 3: production example of sugar solution from bagasse (Steps (1) to (5) were performed)
The hydrolyzed sugar solution obtained in step (4) in example 1 was subjected to solid-liquid separation by centrifugation (2,000 rpm, manufactured by , , a rattan, religious purification), filtered through a microfiltration membrane (a 0.05 μm pore diameter vdf membrane, manufactured by ミリポア), passed through a reverse osmosis membrane (a cross-linked wholly aromatic polyamide reverse osmosis membrane UTC80, manufactured by レ), and the sugar solution was recovered as a concentrated solution, and water was recovered as a permeated solution by being concentrated to 1 in 4 minutes of the raw water.
Next, the steps (1) to (4) were carried out under the same conditions as in example 1, and water recovered as a permeate of the reverse osmosis membrane was used for the addition of water in step (3) (step (5)). The sugar, acetic acid, and aromatic compound concentrations of the sugar solution obtained in step (4) were measured, and the weights of acetic acid and coumaric acid were determined based on 100g of the obtained glucose, and the results are shown in table 3.
[ Table 3]
As can be seen from table 3, the weight of acetic acid and coumaric acid with respect to glucose was relatively reduced in examples 1 to 3, as compared with all of comparative examples 1 to 3 in which steps (1) to (4) were not performed. In particular, as is clear from the comparison results of comparative example 2 and examples 1 and 2, if the weight ratio (dry weight%) of the powder biomass which does not pass through a sieve having a mesh size of 1mm as the pulverization degree of the powder biomass exceeds 50%, the concentration of the fermentation-inhibiting substance in the sugar solution increases. Further, as is clear from the results of examples 1 and 3, the concentration of coumaric acid in the sugar solution was further reduced by performing step (5).
Claims (10)
1. A method for producing a sugar solution, which uses a cellulose-containing biomass as a raw material and comprises the following steps:
step (1): a step of pulverizing the cellulose-containing biomass so that the weight ratio of the cellulose-containing biomass that does not pass through a sieve having a mesh opening of 1mm is 50% or less by dry weight;
step (2): a step of bringing the pulverized biomass obtained in the step (1) into contact with an alkaline aqueous medium to obtain a pretreated biomass;
step (3): adding water to the pretreated biomass obtained in step (2) and performing solid-liquid separation to obtain a cellulose-containing solid component; and
step (4): and (4) hydrolyzing the cellulose-containing solid component obtained in step (3) to obtain a sugar solution.
2. The method for producing a sugar solution according to claim 1, wherein the step (2) is a step of obtaining a pretreated biomass by introducing an alkaline medium into the pulverized biomass obtained in the step (1).
3. The method for producing a sugar solution according to claim 1 or 2, wherein the step (2) is a step of supplying the pulverized biomass obtained in the step (1) and the alkaline medium to a filter and allowing the pulverized biomass and the alkaline medium to pass through the filter.
4. The method for producing a sugar solution according to any one of claims 1 to 3, wherein the alkaline aqueous medium in the step (2) is an aqueous medium containing sodium hydroxide and/or potassium hydroxide.
5. The method for producing a sugar solution according to any one of claims 1 to 4, wherein the solid-liquid separation in the step (3) is squeezing.
6. The method for producing a sugar solution according to any one of claims 1 to 5, wherein the cellulose-containing biomass is bagasse.
7. The method for producing a sugar solution according to any one of claims 1 to 6, further comprising the following step (5): and (3) filtering the sugar solution obtained in step (4) with a nanofiltration membrane or a reverse osmosis membrane to recover the sugar solution as a non-permeate, and reusing the permeate as the water added to the pretreated biomass in step (3).
8. A cellulose-containing solid component comprising a pulverized cellulose-containing biomass and an alkaline aqueous medium, wherein the pulverized cellulose-containing biomass has a weight ratio of not passing through a sieve having a sieve opening of 1mm of 50% by weight or less in terms of dry weight, and the cellulose-containing solid component has a water content of 50% by weight or more and less than 70% by weight.
9. The cellulose-containing solid component of claim 8, the cellulose-containing biomass being bagasse.
10. A pulverized cellulose-containing biomass which does not pass through a sieve having a mesh size of 1mm, wherein the weight ratio of the pulverized cellulose-containing biomass to the dry weight is 40-50%.
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| JP2010220512A (en) * | 2009-03-23 | 2010-10-07 | Jfe Engineering Corp | Pretreatment method for enzymic hydrolysis treatment of herbaceous biomass, and method for producing ethanol by using herbaceous biomass as raw material |
| JP2013220067A (en) * | 2012-04-17 | 2013-10-28 | National Agriculture & Food Research Organization | Enzymatic saccharification method for cellulosic biomass raw material |
| CN105473729A (en) * | 2013-08-22 | 2016-04-06 | 东丽株式会社 | Method for manufacturing sugar solution |
| CN105745331A (en) * | 2013-12-27 | 2016-07-06 | 东丽株式会社 | Method for producing sugar solution |
| CN108884479A (en) * | 2016-03-29 | 2018-11-23 | 东丽株式会社 | The manufacturing method of liquid glucose |
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| CA2868572C (en) * | 2012-03-29 | 2015-04-28 | Mitsubishi Heavy Industries Mechatronics Systems, Ltd. | Biomass processing system, saccharide solution production method using biomass feedstock, alcohol production method |
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| JP2010220512A (en) * | 2009-03-23 | 2010-10-07 | Jfe Engineering Corp | Pretreatment method for enzymic hydrolysis treatment of herbaceous biomass, and method for producing ethanol by using herbaceous biomass as raw material |
| JP2013220067A (en) * | 2012-04-17 | 2013-10-28 | National Agriculture & Food Research Organization | Enzymatic saccharification method for cellulosic biomass raw material |
| CN105473729A (en) * | 2013-08-22 | 2016-04-06 | 东丽株式会社 | Method for manufacturing sugar solution |
| CN105745331A (en) * | 2013-12-27 | 2016-07-06 | 东丽株式会社 | Method for producing sugar solution |
| CN108884479A (en) * | 2016-03-29 | 2018-11-23 | 东丽株式会社 | The manufacturing method of liquid glucose |
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