CN105969822A - Lignocellulose pretreatment method based on ionic liquid - Google Patents
Lignocellulose pretreatment method based on ionic liquid Download PDFInfo
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Abstract
The invention discloses a lignocellulose pretreatment method based on ionic liquid. The lignocellulose pretreatment method includes the concrete steps that 1, an ionic liquid and seawater mixture is used as pretreatment solvent, under protection of nitrogen, lignocellulose and the pretreatment solvent are mixed according to the mass ratio of 1:5-1:25 and stirred for 1-24 h at the temperature of 50-150 DEG C, obtained treating liquid is cooled to room temperature and filtered, filter residue is washed with fresh water or sea water, and the pretreated lignocelluloses residue is obtained after drying; 2, a water solution with the pH value of 4.8 is added into the lignocelluloses residue, cellulase and beta-glucosaccharase are added, and fermentable revertose solution is obtained after enzymolysis reaction. Enzymolysis efficiency of lignocellulose can be effectively reinforced through the pretreatment process, the yield of ermentable revertose is increased, and the method further has the advantages of being environmentally friendly, low in viscosity, easy to operate, low in cost and low in energy consumption and fresh water consumption.
Description
Technical field
The invention belongs to lignocellulose utilize and chemical industry application field, be specifically related to one and utilize sea water
Substitute fresh water for ionic liquid pretreatment lignocellulose and the method for follow-up enzymolysis.
Background technology
Along with outburst and people's growing interest to environmental problem of energy crisis in recent years, biomass etc. can be again
The development and utilization in production-goods source gradually comes into one's own (Science, 2012,337,695).With fossil resource phase
Ratio, biomass resource has many advantages；Such as, its annual production is huge, and fires with biomass for raw material production
Material and industrial chemicals also can effectively reduce CO2Discharge and environmental pollution.Lignocellulose is as on the earth
It is distributed the widest and that content is the abundantest renewable biomass resource and enjoys the concern of people.Lignocellulose master
To be made up of lignin, cellulose and hemicellulose.Lignocellulose can be converted into various through biorefining
The bio-based energy and platform chemicals.But, owing to lignocellulose structure is complicated, cellulose is not only by half
Cellulose and lignin parcel, and its intramolecular and a large amount of hydrogen bond of intermolecular existence, cause its highly crystallized,
Therefore being difficult to directly utilize chemical method or bioanalysis by its Efficient Conversion is bioenergy or platform chemicals, this is tight
Heavily hinder the development and utilization of lignocellulose.It is thus typically necessary to lignocellulose is carried out pretreatment,
Remove partial lignin, reduce cellulose crystallity, destroy its three dimensional structure, thus increase reaction site,
Improve reducing sugar yield (ChemSusChem, 2015,8,3366).
Ionic liquid is owing to having superpower solvability, and the lignocellulose having proved to be a class excellent is pre-
Process solvent (Green Chem, 2013,15,550).While it is true, ionic liquid pretreatment wood fibre
Element need to consume a large amount of precious freshwater resources.Such as, after ionic liquid pretreatment, need to be to pretreated
Lignocellulosic residue is repeatedly washed, to reduce the loss of ionic liquid and survivor ion liquid to follow-up
The negative effect of enzymolysis, and be also required to use a large amount of fresh water in follow-up enzymolysis process
(CN103045700B).It is reported, often produce 1m3Bio-fuel takes around consumption 1.9-5.9m3
Water (ChemSusChem, 2015,8,3823).But, due to population in the world surplus and environmental pollution etc.
The problem that aggravation, Countries and area have occurred serious shortage of fresh water.This undoubtedly can greatly obstacle from
Sub-liquid large-scale application in lignocellulosic biomass refine.
Summary of the invention
The problem existed for prior art, it is an object of the invention to provide one and utilizes sea water replacement light
Water is used for ionic liquid pretreatment lignocellulose and the method for follow-up enzymolysis.
The purpose of the present invention is achieved through the following technical solutions:
A kind of lignocellulose pretreatment method based on ionic liquid, comprises the steps:
(1) with ionic liquid-sea water mixing thing as pre-treatment solvents, under nitrogen protection, lignocellulose
It is that 1:5～1:25 mixes with pre-treatment solvents according to mass ratio, stirs 1～24 hour at 50～150 DEG C,
Gained treatment fluid is then cooled to room temperature, filters, utilizes fresh water or seawer washing filtering residue, and dried acquisition is pre-
Lignocellulosic residue after process；Described ionic liquid is choline or glyoxaline ion liquid；
(2) in above-mentioned lignocellulosic residue, add pH 4.8 aqueous solution, add cellulase and β-
Glucosidase, after enzyme digestion reaction, it is thus achieved that fermentable reducing sugar solution.
Described choline-like ionic liquid is the ionic liquid with choline as cation；Described imidazoles from
Sub-liquid is the ionic liquid with imidazoles as cation.
Choline-like ionic liquid described in step (1) is with choline as cation, amino acid salts or taurine
Salt is anion ion liquid；Described glyoxaline ion liquid is [1-ethyl-3-methylimidazole] [acetic acid
Salt].
Described amino acid salts be glycinate, alanine salt, serine salt, threonine salt, valine salt,
Leucine salt, isoleucine salt, methionine salt, phenylalanine salt, proline salt, tryptophan salt, bad ammonia
Hydrochlorate, arginine salt or histidine salt.
The condition of the enzyme digestion reaction described in step (2) is hunting speed 150～250r/min, reaction temperature
40～60 DEG C, the response time 12～72h.
Lignocellulose described in step (1) is rice straw, corn straw, corn cob, wheat straw
Stalk or bagasse.
Ionic liquid described in step (1)-sea water mixing thing intermediate ion content liquid is 5～95wt%.
During described ionic liquid-sea water mixing thing intermediate ion content liquid >=50wt%, gained treatment fluid exists
Adding diluted before filtration, described diluent is sea water or fresh water.
The volume of described diluent is 1～3 times of pre-treatment solvents volume.
PH 4.8 aqueous solution described in step (2) is 50mmol/L, pH 4.8 citrate buffer
Or utilize pH 4.8 sea water that citric acid regulates.
Cellulase consumption described in step (2) is 5～15FPU/g pretreated lignocellulose
Residue；Described beta-glucosidase enzyme dosage is 15～40CBU/g pretreated lignocellulosic residue.
Lignocellulosic residue described in step (2) is 1～10 with the solid-to-liquid ratio of pH 4.8 aqueous solution
mg/mL。
Described cellulase is the commercial fibers element enzyme deriving from Trichoderma spp. (Trichoderma sp.)；Institute
The beta-glucosidase stated is the commercialization beta-glucosidase deriving from aspergillus niger (Aspergillus niger)
Enzyme.
The present invention compared with prior art, has the advantage that
1) utilize cheap, be easy to get and fresh water that sea water alternate resources that storage is huge is limited is for ionic liquid
Body preprocessing lignocellulose and follow-up enzymolysis, will significantly reduce lignocellulosic biomass essence
Fresh water consumption in refining, thus the problem overcoming " robbing water with people ".
2) containing substantial amounts of moisture in the pre-treatment solvents used by the present invention, therefore this pretreating process has super
Strong resistance to water, therefore before pretreatment, it is not necessary to ionic liquid and lignocellulose are dried,
Simplify technical process, and significantly reduce energy consumption.
3) ionic liquid that the present invention is of a relatively high in price, viscosity is bigger adds a certain amount of, cheap,
Low viscous sea water, can not only effectively reduce system viscosity, conveniently operation, and will greatly
Reduce the consumption of ionic liquid, thus reduce cost.Meanwhile, add a certain amount of sea water can have
The pre-processing intensity (ionic liquid alkalescence) of effect regulation ionic liquid, lowers the loss of hemicellulose,
Improve reducing sugar yield.
4) present invention utilizes ionic liquid-sea water compound as lignocellulose pre-treatment solvents, significantly increases
The strong enzymolysis of follow-up lignocellulose, improves reducing sugar yield.
Detailed description of the invention
Further illustrate the present invention by embodiment, but be not limited only to embodiment.
Embodiment 1
80wt% [choline] [arginine salt]-sea water mixing thing pretreatment wheat stalk
A) pretreatment: accurate weighing 150mg wheat stalk powder (250～420 μm) and 3g [choline] [essence
Propylhomoserin salt] (ionic liquid content is 80wt% to-sea water mixing thing；Ionic liquid according to document [Green Chem,
2012,14,304] described method synthesis), it is collectively disposed in 15mL triangular flask, under nitrogen protection, in
Stir 6 hours at 90 DEG C；It is then cooled to room temperature, adds the deionized water of 1 times of pre-treatment solvents volume
Dilution, filter, then with the deionized water wash filtering residue 5 times of 3 times of volumes, filtering residue the most i.e. obtains pre-
Wheat stalk residue after process.
B) enzymolysis: accurate weighing above-mentioned wheat stalk residue 20mg, is placed in the triangular flask of 50mL,
Add 7mL citrate buffer (50mmol/L, pH 4.8), 11FPU/g derives from the fiber of Trichoderma spp.
Element enzyme and 25CBU/g derive from the beta-glucosidase (being all purchased from Novozymes Company) of aspergillus niger, seal
It is placed in 200r/min, the constant temperature oscillator of 50 DEG C and reacts.After 24h, reducing sugar yield is 71% (profit
With DNS method measure concentration of reduced sugar, Pure Appl.Chem.1987,59,257；Based on little before pretreatment
The theoretical yield of reducing sugar in Wheat Straw, calculates reducing sugar yield).
Embodiment 2
80wt% [choline] [glycinate]-sea water mixing thing pretreatment wheat stalk
A) pretreatment: accurate weighing 150mg wheat stalk powder (250～420 μm) and 3g [choline] are [sweet
Propylhomoserin salt]-sea water mixing thing (ionic liquid content is 80wt%), it is collectively disposed in 15mL triangular flask,
Under nitrogen protection, stir 6 hours at 90 DEG C；It is then cooled to room temperature, adds 1 times of pretreatment molten
The deionized water dilution of agent volume, filter, then with the deionized water wash filtering residue 5 times of 3 times of volumes, filtering residue
The most i.e. obtain pretreated wheat stalk residue.
B) enzymolysis: accurate weighing above-mentioned wheat stalk residue 20mg, is placed in the triangular flask of 50mL,
Add 7mL citrate buffer (50mmol/L, pH 4.8), 11FPU/g derives from the fiber of Trichoderma spp.
Element enzyme and 25CBU/g derive from the beta-glucosidase (being all purchased from Novozymes Company) of aspergillus niger, seal
It is placed in 200r/min, the constant temperature oscillator of 50 DEG C and reacts.After 24h, reducing sugar yield is 66%.
Embodiment 3
80wt% [choline] [lysinate]-sea water mixing thing pretreatment wheat stalk
A) pretreatment: accurate weighing 150mg wheat stalk powder (250～420 μm) and 3g [choline] [rely
Propylhomoserin salt]-sea water mixing thing (ionic liquid content is 80wt%), it is collectively disposed in 15mL triangular flask,
Under nitrogen protection, stir 6 hours at 90 DEG C；It is then cooled to room temperature, adds 1 times of pretreatment molten
The deionized water dilution of agent volume, filter, then with the deionized water wash filtering residue 5 times of 3 times of volumes, filtering residue
The most i.e. obtain pretreated wheat stalk residue.
B) enzymolysis: accurate weighing above-mentioned wheat stalk residue 20mg, is placed in the triangular flask of 50mL,
Add 7mL citrate buffer (50mmol/L, pH 4.8), 11FPU/g derives from the fiber of Trichoderma spp.
Element enzyme and 25CBU/g derive from the beta-glucosidase (being all purchased from Novozymes Company) of aspergillus niger, seal
It is placed in 200r/min, the constant temperature oscillator of 50 DEG C and reacts.After 48h, reducing sugar yield is 63%.
Embodiment 4
80wt% [choline] [phenylalanine salt]-sea water mixing thing pretreatment wheat stalk
A) pretreatment: accurate weighing 150mg wheat stalk powder (250～420 μm) and 3g [choline] [benzene
Alanine salt]-sea water mixing thing (ionic liquid content is 80wt%), it is collectively disposed in 15mL triangular flask,
Under nitrogen protection, stir 6 hours at 90 DEG C；It is then cooled to room temperature, adds 1 times of pretreatment molten
The deionized water dilution of agent volume, filter, then with the deionized water wash filtering residue 5 times of 3 times of volumes, filtering residue
The most i.e. obtain pretreated wheat stalk residue.
B) enzymolysis: accurate weighing above-mentioned wheat stalk residue 20mg, is placed in the triangular flask of 50mL,
Add 7mL citrate buffer (50mmol/L, pH 4.8), 11FPU/g derives from the fiber of Trichoderma spp.
Element enzyme and 25CBU/g derive from the beta-glucosidase (being all purchased from Novozymes Company) of aspergillus niger, seal
It is placed in 200r/min, the constant temperature oscillator of 50 DEG C and reacts.After 48h, reducing sugar yield is 63%.
Embodiment 5
80wt% [choline] [valine salt]-sea water mixing thing pretreatment wheat stalk
A) pretreatment: accurate weighing 150mg wheat stalk powder (250～420 μm) and 3g [choline] [figured silk fabrics
Propylhomoserin salt]-sea water mixing thing (ionic liquid content is 80wt%), it is collectively disposed in 15mL triangular flask,
Under nitrogen protection, stir 6 hours at 90 DEG C；It is then cooled to room temperature, adds 1 times of pretreatment molten
The deionized water dilution of agent volume, filter, then with the deionized water wash filtering residue 5 times of 3 times of volumes, filtering residue
The most i.e. obtain pretreated wheat stalk residue.
B) enzymolysis: accurate weighing above-mentioned wheat stalk residue 20mg, is placed in the triangular flask of 50mL,
Add 7mL citrate buffer (50mmol/L, pH 4.8), 11FPU/g derives from the fiber of Trichoderma spp.
Element enzyme and 25CBU/g derive from the beta-glucosidase (being all purchased from Novozymes Company) of aspergillus niger, seal
It is placed in 200r/min, the constant temperature oscillator of 50 DEG C and reacts.After 48h, reducing sugar yield is 64%.
Embodiment 6
80wt% [choline] [proline salt]-sea water mixing thing pretreatment wheat stalk
A) pretreatment: accurate weighing 150mg wheat stalk powder (250～420 μm) and 3g [choline] [dried meat
Propylhomoserin salt]-sea water mixing thing (ionic liquid content is 80wt%), it is collectively disposed in 15mL triangular flask,
Under nitrogen protection, stir 6 hours at 90 DEG C；It is then cooled to room temperature, adds 1 times of pretreatment molten
The deionized water dilution of agent volume, filter, then with the deionized water wash filtering residue 5 times of 3 times of volumes, filtering residue
The most i.e. obtain pretreated wheat stalk residue.
B) enzymolysis: accurate weighing above-mentioned wheat stalk residue 20mg, is placed in the triangular flask of 50mL,
Add 7mL citrate buffer (50mmol/L, pH 4.8), 11FPU/g derives from the fiber of Trichoderma spp.
Element enzyme and 25CBU/g derive from the beta-glucosidase (being all purchased from Novozymes Company) of aspergillus niger, seal
It is placed in 200r/min, the constant temperature oscillator of 50 DEG C and reacts.After 48h, reducing sugar yield is 66%.
Embodiment 7
80wt% [choline] [taurate]-sea water mixing thing pretreatment wheat stalk
A) pretreatment: accurate weighing 150mg wheat stalk powder (250～420 μm) and 3g [choline] [cattle
Sulfonate]-sea water mixing thing (ionic liquid content is 80wt%), it is collectively disposed in 15mL triangular flask,
Under nitrogen protection, stir 6 hours at 90 DEG C；It is then cooled to room temperature, adds 1 times of pretreatment molten
The deionized water dilution of agent volume, filter, then with the deionized water wash filtering residue 5 times of 3 times of volumes, filtering residue
The most i.e. obtain pretreated wheat stalk residue.
B) enzymolysis: accurate weighing above-mentioned wheat stalk residue 20mg, is placed in the triangular flask of 50mL,
Add 7mL citrate buffer (50mmol/L, pH 4.8), 11FPU/g derives from the fiber of Trichoderma spp.
Element enzyme and 25CBU/g derive from the beta-glucosidase (being all purchased from Novozymes Company) of aspergillus niger, seal
It is placed in 200r/min, the constant temperature oscillator of 50 DEG C and reacts.After 72h, reducing sugar yield is 71%.
Embodiment 8
80wt% [1-ethyl-3-methylimidazole] [acetate]-sea water mixing thing pretreatment wheat stalk
A) pretreatment: accurate weighing 150mg wheat stalk powder (250～420 μm) and 3g [1-second
Base-3-Methylimidazole .] [acetate]-sea water mixing thing (ionic liquid content is 80wt%), it is collectively disposed at 15
In mL triangular flask, under nitrogen protection, stir 6 hours at 90 DEG C；It is then cooled to room temperature, adds
Enter the deionized water dilution of 1 times of pre-treatment solvents volume, filter, then the deionized water wash with 3 times of volumes
Filtering residue 5 times, filtering residue the most i.e. obtains pretreated wheat stalk residue.
B) enzymolysis: accurate weighing above-mentioned wheat stalk residue 20mg, is placed in the triangular flask of 50mL,
Add 7mL citrate buffer (50mmol/L, pH 4.8), 11FPU/g derives from the fiber of Trichoderma spp.
Element enzyme and 25CBU/g derive from the beta-glucosidase (being all purchased from Novozymes Company) of aspergillus niger, seal
It is placed in 200r/min, the constant temperature oscillator of 50 DEG C and reacts.After 72h, reducing sugar yield is 43%.
Embodiment 9
50wt% [1-ethyl-3-methylimidazole] [acetate]-sea water mixing thing pretreatment wheat stalk
A) pretreatment: accurate weighing 150mg wheat stalk powder (250～420 μm) and 3g [1-second
Base-3-Methylimidazole .] [acetate]-sea water mixing thing (ionic liquid content is 50wt%), it is collectively disposed at 15
In mL triangular flask, under nitrogen protection, stir 1.5 hours at 150 DEG C；It is then cooled to room temperature,
Add the deionized water dilution of 1 times of pre-treatment solvents volume, filter, then wash with the deionization of 3 times of volumes
Washing filtering residue 5 times, filtering residue the most i.e. obtains pretreated wheat stalk residue.
B) enzymolysis: accurate weighing above-mentioned wheat stalk residue 20mg, is placed in the triangular flask of 50mL,
Add 7mL citrate buffer (50mmol/L, pH 4.8), 11FPU/g derives from the fiber of Trichoderma spp.
Element enzyme and 25CBU/g derive from the beta-glucosidase (being all purchased from Novozymes Company) of aspergillus niger, seal
It is placed in 200r/min, the constant temperature oscillator of 50 DEG C and reacts.After 72h, reducing sugar yield is 55%.
Embodiment 10
20wt% [choline] [arginine salt]-sea water mixing thing pretreatment wheat stalk
A) pretreatment: accurate weighing 150mg wheat stalk powder (250～420 μm) and 3g [choline] [essence
Propylhomoserin salt]-sea water mixing thing (ionic liquid content is 20wt%), it is collectively disposed in 15mL triangular flask,
Under nitrogen protection, stir 6 hours at 90 DEG C；It is then cooled to room temperature, filters, then with 3 times of bodies
Long-pending deionized water wash filtering residue 5 times, filtering residue the most i.e. obtains pretreated wheat stalk residue.
B) enzymolysis: accurate weighing above-mentioned wheat stalk residue 20mg, is placed in the triangular flask of 50mL,
Add 7mL citrate buffer (50mmol/L, pH 4.8), 11FPU/g derives from the fiber of Trichoderma spp.
Element enzyme and 25CBU/g derive from the beta-glucosidase (being all purchased from Novozymes Company) of aspergillus niger, seal
It is placed in 200r/min, the constant temperature oscillator of 50 DEG C and reacts.After 48h, reducing sugar yield is 77%.
Embodiment 11
50wt% [choline] [arginine salt]-sea water mixing thing pretreatment wheat stalk
A) pretreatment: accurate weighing 150mg wheat stalk powder (250～420 μm) and 3g [choline] [essence
Propylhomoserin salt]-sea water mixing thing (ionic liquid content is 50wt%), it is collectively disposed in 15mL triangular flask,
Under nitrogen protection, stir 6 hours at 90 DEG C；It is then cooled to room temperature, adds 1 times of pretreatment molten
The sea water dilution of agent volume, filter, then with the seawer washing filtering residue 5 times of 3 times of volumes, filtering residue is after drying
I.e. obtain pretreated wheat stalk residue.
B) enzymolysis: (265mg, this residue is wheat stalk residue by above-mentioned all of wheat stalk residue
And the mixture of sea salt；Wherein, wheat stalk mass of residue is about 80mg), it is placed in the triangle of 100mL
In Ping, add 28mL sea water (pH 4.8, pH is regulated) by citric acid, 11FPU/g derives from Trichoderma spp.
Cellulase and 25CBU/g derive from the beta-glucosidase (being all purchased from Novozymes Company) of aspergillus niger,
Seal to be placed in 200r/min, the constant temperature oscillator of 50 DEG C and react.After 48h, reducing sugar yield is 60%.
Embodiment 12
50wt% [choline] [arginine salt]-sea water mixing thing pretreated water rice straw
A) pretreatment: accurate weighing 150mg rice straw powder (250～420 μm) and 3g [choline] [essence
Propylhomoserin salt]-sea water mixing thing (ionic liquid content is 50wt%), it is collectively disposed in 15mL triangular flask,
Under nitrogen protection, stir 6 hours at 90 DEG C；It is then cooled to room temperature, adds 1 times of pretreatment molten
The sea water dilution of agent volume, filter, then with the seawer washing filtering residue 5 times of 3 times of volumes, filtering residue is after drying
I.e. obtain pretreated rice straw residue.
B) enzymolysis: by above-mentioned all of rice straw residue (249mg；Wherein, rice straw residue matter
Amount is about 71mg), it is placed in the triangular flask of 100mL, (pH 4.8, pH passes through to add 25mL sea water
Citric acid regulates), 11FPU/g derive from the cellulase of Trichoderma spp. and 25CBU/g derive from the β of aspergillus niger-
Glucosidase (is all purchased from Novozymes Company), seals and is placed on 200r/min, the constant temperature oscillation of 50 DEG C
Device reacts.After 48h, reducing sugar yield is 54%.
Embodiment 13
50wt% [choline] [arginine salt]-sea water mixing thing pretreatment corn cob
A) pretreatment: accurate weighing 150mg corncob powder (250～420 μm) and 3g [choline] [essence
Propylhomoserin salt]-sea water mixing thing (ionic liquid content is 50wt%), it is collectively disposed in 15mL triangular flask,
Under nitrogen protection, stir 6 hours at 90 DEG C；It is then cooled to room temperature, adds 1 times of pretreatment molten
The sea water dilution of agent volume, filter, then with the seawer washing filtering residue 5 times of 3 times of volumes, filtering residue is after drying
I.e. obtain pretreated corn cob residue.
B) enzymolysis: by above-mentioned all of corn cob residue (270mg；Wherein, corn cob mass of residue is about
For 91mg), it is placed in the triangular flask of 100mL, (pH 4.8, pH passes through Fructus Citri Limoniae to add 32mL sea water
Acid regulation), 11FPU/g derives from the cellulase of Trichoderma spp. and 25CBU/g derives from the β-Portugal of aspergillus niger
Polyglycoside enzyme (is all purchased from Novozymes Company), seals and is placed on 200r/min, the constant temperature oscillator of 50 DEG C
Middle reaction.After 48h, reducing sugar yield is 72%.
Embodiment 14
50wt% [choline] [arginine salt]-sea water mixing thing pretreatment bagasse
A) pretreatment: accurate weighing 150mg bagasse powder (250～420 μm) and 3g [choline] [essence
Propylhomoserin salt]-sea water mixing thing (ionic liquid content is 50wt%), it is collectively disposed in 15mL triangular flask,
Under nitrogen protection, stir 6 hours at 90 DEG C；It is then cooled to room temperature, adds 1 times of pretreatment molten
The sea water dilution of agent volume, filter, then with the seawer washing filtering residue 5 times of 3 times of volumes, filtering residue is after drying
I.e. obtain pretreated bagasse residue.
B) enzymolysis: by above-mentioned all of bagasse residue (241mg；Wherein, bagasse mass of residue is about
For 65mg), it is placed in the triangular flask of 100mL, (pH 4.8, pH passes through Fructus Citri Limoniae to add 25mL sea water
Acid regulation), 11FPU/g derives from the cellulase of Trichoderma spp. and 25CBU/g derives from the β-Portugal of aspergillus niger
Polyglycoside enzyme (is all purchased from Novozymes Company), seals and is placed on 200r/min, the constant temperature oscillator of 50 DEG C
Middle reaction.After 48h, reducing sugar yield is 55%.
Embodiment 15
50wt% [choline] [arginine salt]-sea water mixing thing pretreatment corn straw
A) pretreatment: accurate weighing 150mg corn straw powder (250～420 μm) and 3g [choline] [essence
Propylhomoserin salt]-sea water mixing thing (ionic liquid content is 50wt%), it is collectively disposed in 15mL triangular flask,
Under nitrogen protection, stir 6 hours at 90 DEG C；It is then cooled to room temperature, adds 1 times of pretreatment molten
The sea water dilution of agent volume, filter, then with the seawer washing filtering residue 5 times of 3 times of volumes, filtering residue is after drying
I.e. obtain pretreated corn straw residue.
B) enzymolysis: by above-mentioned all of corn straw residue (198mg；Wherein, corn straw residue matter
Amount is about 62mg), it is placed in the triangular flask of 100mL, (pH 4.8, pH passes through to add 25mL sea water
Citric acid regulates), 11FPU/g derive from the cellulase of Trichoderma spp. and 25CBU/g derive from the β of aspergillus niger-
Glucosidase (is all purchased from Novozymes Company), seals and is placed on 200r/min, the constant temperature oscillation of 50 DEG C
Device reacts.After 48h, reducing sugar yield is 60%.
Comparative example 1
The most pretreated wheat stalk enzymolysis
Accurate weighing 20mg the most pretreated wheat stalk powder (250～420 μm), is placed in 50mL
Triangular flask in, add 7mL sea water (pH 4.8, pH is regulated) by citric acid, 11FPU/g derives from
The cellulase of Trichoderma spp. and 25CBU/g derive from the beta-glucosidase of aspergillus niger and (are all purchased from Novi's letter public affairs
Department), seal to be placed in 200r/min, the constant temperature oscillator of 50 DEG C and react.After 48h, reducing sugar is received
Rate is 15%.
Comparative example 2
The most pretreated wheat stalk enzymolysis
Accurate weighing 20mg the most pretreated wheat stalk powder (250～420 μm), is placed in 50mL
Triangular flask in, add 7mL citrate buffer (50mmol/L, pH 4.8), 11FPU/g source
The beta-glucosidase deriving from aspergillus niger in the cellulase of Trichoderma spp. and 25CBU/g (is all purchased from Novi's letter
Company), seal to be placed in 200r/min, the constant temperature oscillator of 50 DEG C and react.After 48h, reducing sugar
Yield is 15%.
Comparative example 3
The most pretreated rice straw enzymolysis
Accurate weighing 20mg the most pretreated rice straw powder (250～420 μm), is placed in 50mL
Triangular flask in, add 7mL sea water (pH 4.8, pH is regulated) by citric acid, 11FPU/g derives from
The cellulase of Trichoderma spp. and 25CBU/g derive from the beta-glucosidase of aspergillus niger and (are all purchased from Novi's letter public affairs
Department), seal to be placed in 200r/min, the constant temperature oscillator of 50 DEG C and react.After 48h, reducing sugar is received
Rate is 17%.
Comparative example 4
The most pretreated corn cob enzymolysis
The most pretreated corncob powder of accurate weighing 20mg (250～420 μm), is placed in 50mL
Triangular flask in, add 7mL sea water (pH 4.8, pH is regulated) by citric acid, 11FPU/g derives from
The cellulase of Trichoderma spp. and 25CBU/g derive from the beta-glucosidase of aspergillus niger and (are all purchased from Novi's letter public affairs
Department), seal to be placed in 200r/min, the constant temperature oscillator of 50 DEG C and react.After 48h, reducing sugar is received
Rate is 14%.
Claims (10)
1. a lignocellulose pretreatment method based on ionic liquid, it is characterised in that include as follows
Step:
(1) with ionic liquid-sea water mixing thing as pre-treatment solvents, under nitrogen protection, lignocellulose
It is that 1:5～1:25 mixes with pre-treatment solvents according to mass ratio, stirs 1～24 hour at 50～150 DEG C,
Gained treatment fluid is then cooled to room temperature, filters, utilizes fresh water or seawer washing filtering residue, and dried acquisition is pre-
Lignocellulosic residue after process；Described ionic liquid is choline or glyoxaline ion liquid；
(2) in above-mentioned lignocellulosic residue, add pH 4.8 aqueous solution, add cellulase and β-
Glucosidase, after enzyme digestion reaction, it is thus achieved that fermentable reducing sugar solution.
Method the most according to claim 1, it is characterised in that the choline described in step (1) from
Sub-liquid is with choline as cation, and amino acid salts or taurate are anion ion liquid；Described
Glyoxaline ion liquid is [1-ethyl-3-methylimidazole] [acetate].
Method the most according to claim 2, it is characterised in that described amino acid salts be glycinate,
Alanine salt, serine salt, threonine salt, valine salt, leucine salt, isoleucine salt, methionine
Salt, phenylalanine salt, proline salt, tryptophan salt, lysinate, arginine salt or histidine salt.
4. according to the method described in claim 1 or 2 or 3, it is characterised in that described in step (2)
The condition of enzyme digestion reaction is hunting speed 150～250r/min, reaction temperature 40～60 DEG C, the response time
12～72h.
Method the most according to claim 4, it is characterised in that the wooden fibre described in step (1)
Dimension element is rice straw, corn straw, corn cob, wheat stalk or bagasse.
Method the most according to claim 5, it is characterised in that the ionic liquid described in step (1)
Body-sea water mixing thing intermediate ion content liquid is 5～95wt%；In described ionic liquid-sea water mixing thing
During ionic liquid content >=50wt%, gained treatment fluid adds diluted before filtration, described dilute
Releasing liquid is sea water or fresh water；The volume of described diluent is 1～3 times of pre-treatment solvents volume.
7. according to the method described in claim 1～6 any one, it is characterised in that in step (2)
Described pH 4.8 aqueous solution is 50mmol/L, pH 4.8 citrate buffer or utilize citric acid to regulate
PH 4.8 sea water.
Method the most according to claim 7, it is characterised in that the wooden fibre described in step (2)
Dimension element residue is 1～10mg/mL with the solid-to-liquid ratio of pH 4.8 aqueous solution.
Method the most according to claim 8, it is characterised in that the cellulose described in step (2)
Enzyme dosage is 5～15FPU/g pretreated lignocellulosic residue；Described beta-glucosidase enzyme dosage
It is 15～40CBU/g pretreated lignocellulosic residue.
Method the most according to claim 9, it is characterised in that described cellulase is for deriving from
The commercial fibers element enzyme of Trichoderma spp. (Trichoderma sp.)；Described beta-glucosidase is black for deriving from
The commercialization beta-glucosidase of aspergillosis (Aspergillus niger).
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| JP2018065984A (en) * | 2016-10-18 | 2018-04-26 | パナソニックＩｐマネジメント株式会社 | Ionic liquid and method for dissolving cellulose using the same |
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| CN107467382A (en) * | 2017-10-11 | 2017-12-15 | 广西南宁荣威德新能源科技有限公司 | A kind of beef cattle special feed and preparation method thereof |
| CN109851679A (en) * | 2019-02-18 | 2019-06-07 | 中国科学院过程工程研究所 | A kind of arrest method degraded when cellulose dissolution in ion liquid system |
| CN110066840A (en) * | 2019-03-08 | 2019-07-30 | 华南农业大学 | A kind of method that hydro-thermal-microwave coupled ion liquid two-step method lignocellulosic material improves enzymolysis efficiency |
| CN110241141A (en) * | 2019-07-29 | 2019-09-17 | 江南大学 | A kind of method that preprocessing lignocellulose improves anaerobic fermentation gas production rate |
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