CN106282265A - A kind of difunctionalization ionic liquid pretreatment biomass improve the method for enzymolysis efficiency - Google Patents
A kind of difunctionalization ionic liquid pretreatment biomass improve the method for enzymolysis efficiency Download PDFInfo
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Abstract
The present invention provides a kind of method that difunctionalization ionic liquid pretreatment biomass improve enzymolysis efficiency, relates to the agricultural-forestry biomass utilization of resources and processing technology field.The method first with containing basic group and containing the difunctionalization ionic liquid of sulfonic acid anion base as solvent, after at a certain temperature protista matter being carried out pretreatment certain time, it is separated by filtration and cleaning obtains residue, the most i.e. obtain pretreated protista matter.With the protista matter after process as substrate, utilize cellulase that it is carried out enzymolysis, finally obtain the sugar liquid based on glucose and xylose.The present invention can not only effectively strengthen the enzymolysis efficiency of protista matter, improves fermentable reducing sugar yield, and relative to system safety non-toxics such as conventional organic solvents, experimental facilities is required low, ionic liquid reusable edible.
Description
Technical field
The present invention relates to the agricultural-forestry biomass utilization of resources and processing technology field, invent a kind of difunctionalization ion
Liquid pretreatment protista matter improves the new method of enzymolysis efficiency.
Background technology
Along with shortage and the rising of price of the non-renewable resources such as oil and coal, and people are dirty to environment
Dye concern so that be only second to coal, oil and natural gas and occupy world energy sources total quantity consumed the 4th
Biomass energy and receive much concern.Biomass energy, as a kind of clean and reproducible energy, is unique
Alternative fossil energy changes into gaseous state, liquid and solid fuel and other industrial chemicals or the carbon of product
Resource.Biomass, as low-grade energy form, are translated into high-grade liquid fuel, are biological
The of paramount importance research direction of energy field.In the conversion process of lignocellulose, life based on sugar platform
Thing transformation technology is owing to reaction condition is gentle, advantages of environment protection enjoys favor.Lignocellulosic material by
Cellulose, hemicellulose and lignin three parts composition.Hemicellulose is combined in fibre as molecule adhesive
Between dimension element and lignin, and the network structure that lignin has, surround as support frame and add set fibre
Dimension element and hemicellulose [Bioresource Technology, 1996,55 (1): 1-33].The network structure of lignin and
The hydrolysis and saccharification of cellulose all can be had a huge impact by the degree of crystallinity of cellulose etc., at biological utilisation fiber
During element, for making microorganism be more easily utilized cellulose, it is necessary to substrate is carried out pretreatment, with degraded
The network structure of lignin, improve to the utilization ratio of cellulose [Biotechnology and Bioengineering,
1995,45(4):328-336]。
Traditional biomass pre-treating method is such as: low-kappa number, oxygenation pretreatment, steam explosion method, hot-water pretreatment,
The quick-fried method of ammonia, but easily produce micro-owing to these methods all exist intrinsic shortcoming, such as low-kappa number method
Biofermentation mortifier, adds the difficulty of subsequent treatment, additionally, the corrosion that diluted acid is to pre-pretreatment unit
Property be also to restrict one of its industrialized key factor；Alkali processes the degree of crystallinity being difficult to destroy lignocellulose,
Compared with amorphous cellulose, its enzymolysis time is the longest；Steam explosion is higher to equipment requirements, equipment
Put into big, constrain scale application.
In recent years, ionic liquid, with the advantage of its uniqueness, becomes the excellent selection of biomass pre-treatment system
[Chem.Commun.,2011,47:1405-1421].Compared with traditional chemical solvent, ionic liquid has
There is a following characteristics: the vapour pressure that (1) is extremely low, it is not easy to volatilization；(2) there is wider liquid journey scope (from low
In or close to room temperature to 300 DEG C), preferable chemical stability and wider electrochemically stable potential window；(3)
By design zwitterion, its dissolubility to inorganic matter, water, Organic substance and polymer of scalable.At present
For the ionic liquid application in biomass pre-treatment field, mainly apply the super solvability of ionic liquid,
Lignin as much as possible is extracted from biomass or separates, because the existence of lignin one side
Face be network structure hinder microorganism close to cellulose and hemicellulose, on the other hand lignin can cause
The ineffective adsorption of enzyme so that enzyme live reduce and enzymolysis cost raise [ChemSusChem, 2013,
6(5):919–927].But while removing lignin, wish reservation cellulose as much as possible and half fiber,
So be conducive to more effectively utilizing the cellulose in biomass and hemicellulose.From existing achievement in research
See, 1,3-dimethylimidazolium methylsulfate (MMim MeSO4)[J.Wood Chem.
Technol.,2007,27,23–33],1-butyl-3-methylimidazolium methylsulfate(Bmim
MeSO4)[J.Wood Chem.Technol.,2007,27,23–33],1-butyl-3-methylimidazolium
acesulfamate(Bmim Ace)[Green Chem.,2011,13,3124-3136],
1-ethyl-3-methylimidazolium acesulfamate(Emim Ace)[Green Chem.,2011,13,
3124-3136]and 1-ethyl-3-methylimidazolium xylenesulfonate(Emim ABS)[Green
Chem.,2009,11,339–345].Lignin is demonstrated preferably by the ionic liquid containing sulfonic acid anion base
Dissolve and extraction effect, but due to ionic liquid be easy to water suction, and sulfonic acid anion base have water with
And under conditions of heating, very solution is hydrolyzed into acidic-group, and such cellulose and hemicellulose are at acid condition
Under be susceptible to degraded.And the invention provides a kind of difunctionalization ionic liquid pretreatment protista matter and carry
The new method of high enzymolysis efficiency, the difunctionalization ionic liquid that basic group and sulfonic acid group are combined is permissible
Effectively remove lignin and reservation cellulose as much as possible and hemicellulose.And relative to traditional
Preprocess method, treatment conditions are gentle, require low to experimental facilities, and the advantage such as ionic liquid is capable of circulation.
Summary of the invention
It is an object of the invention to provide a kind of difunctionalization ionic liquid pretreatment biomass and improve enzymolysis efficiency
Method, the present invention by the following technical solutions:
(1) protista matter drying, pulverizing, its mean diameter controls at 20-200 mesh；
(2) with difunctionalization ionic liquid as pre-treatment solvents, according to the mass ratio of biomass and ionic liquid it is
1:2.5～25 mixing biomass and ionic liquid, stir 0.5～36h at room temperature～200 DEG C, be subsequently cooled to
Room temperature, adds the water of 2～10 times of ionic liquid volumes, filters, and washs filtering residue, obtains pretreatment after drying
After biomass；
(3) it is 1～10mg/ml to weigh pretreated biomass material and join citrate according to solid-to-liquid ratio
In buffer, then add cellulase according to the ratio of 5～30FPU/g pretreated biomass,
Reacting at 100-300r/min, 40-60 DEG C, enzymolysis time is 0.5-100h, and glucose in solutions yield is remote
Much larger than glucose amount produced by undressed biomass material.
Described difunctionalization ionic liquid is containing sulfonic acid anion base but the most fluorine-containing-CHxSO3 -Anion, its
Middle x=0,1 or 2；And sulfonic acid anion base is containing basic group.
Described basic group is-NHx, wherein X=0,1 or 2.
The described ionic liquid containing sulfonic acid anion base: the preferably alkali ionic liquid containing sulfonic acid anion base
Anion structure as follows:
M1And M2It is independently ,-H ,-CH3,-C2H5, C3-C16Straight chain or branched alkane, alkene
Hydrocarbon, aromatic hydrocarbons, monosubstituted aromatic hydrocarbons, di-substituted aryl hydrocarbon and polysubstituted aromatic hydrocarbons；N is 1 or 2；X is-H ,-CH3、
-C2H5, C3-C8Straight chain or branched alkane, alkene, aromatic hydrocarbons, monosubstituted aromatic hydrocarbons, di-substituted aryl hydrocarbon and
Polysubstituted aromatic hydrocarbons.
The described ionic liquid containing sulfonic acid anion base: described anion is preferably
[Et2NC3SO3],[Et2NC4SO3],[Me2NC4SO3],[Me2NC3SO3],
[n-BuNHC3SO3],[n-BuNHC4SO3],[n-HeNHC3SO3],[i-PrNHC3SO3],[i-PrNHC4SO3],[i
-BuNHC3SO3],[i-BuNHC4SO3],[MeNHC4SO3],[MeNHC3SO3],[EtNHC4SO3] or
[EtNHC3SO3]。
The described ionic liquid containing sulfonic acid anion base: the sun of the described ionic liquid containing sulfonic acid anion base
Ion is that imidazoles organic cation, pyridines organic cation, piperidines organic cation, pyroles have
Machine cation, quaternary ammonium salt organic cation, quaternary phosphonium salt organic cation or choline organic cation.
Described imidazoles organic cation general structure is:
Pyridines organic cation general structure is:
Piperidines organic cation general structure is:
Pyroles organic cation general structure is:
Quaternary ammonium salt organic cation general structure is:
Quaternary phosphonium salt organic cation general structure is:
Choline organic cation structure is:
Wherein, R1-R6It is independently ,-CH3,-C2H5Or C3-C6Straight chain or branched alkane
Or alkene；R7-R10It is independently ,-CH3,-C2H5Or C3-C15Straight chain or branched alkane or
Alkene.
The described ionic liquid containing sulfonic acid anion base: cation is preferably tetramethyl amine (Me4N), tetraethyl
Amine (Et4N), 4-butyl amine (Bu4N), 1-methyl-3-ethyl imidazol(e) (Emim), choline (Ch).
Described difunctionalization ionic liquid, preferred ionic liquid is combined as:
Cation is tetramethyl amine (Me4N), tetraethyl amine (Et4N), 4-butyl amine (Bu4N), 1-methyl
-3-ethyl imidazol(e) (Emim), the one in choline (Ch)；
Anion is [Et2NC3SO3],[Et2NC4SO3],[Me2NC4SO3],[Me2NC3SO3],
[n-BuNHC3SO3],[n-BuNHC4SO3],[n-HeNHC3SO3],[i-PrNHC3SO3],[i-PrNHC4SO3],[i
-BuNHC3SO3],[i-BuNHC4SO3],[MeNHC4SO3],[MeNHC3SO3],[EtNHC4SO3],
[EtNHC3SO3One in].
Described difunctionalization ionic liquid: preferably ionic liquid is [Ch] [Et2NC3SO3],[Ch][Me2NC4SO3],[Ch][n-BuNHC3SO3],[Ch][n-HeNHC3SO3],[Ch][i-PrNHC3SO3],[Ch][i-PrNHC4SO3],[Ch][i-BuNHC3SO3],[Et4N][i-PrNHC3SO3],[Et4N][Me2NC4SO3],[Et4N][Et2NC3SO3],[Et4N][n-BuNHC3SO3],[Et4N][i-BuNHC3SO3],[Me4N][MeNHC3SO3], [Me4
N][MeNHC4SO3], [Emim] [MeNHC3SO3] one of which or several mixture.
Described protista raw material have various trees, agricultural crop straw, farming industry side-product,
One or several mixture in feces of livestock and poultry and energy crop etc., described trees are cork and/or hardwood.
Described protista raw material preferable particle size is: 40-80 mesh；Described pretreatment preferable temperature is:
50-180℃。
After described pretreatment, protista matter drying means is lyophilization or heat drying；Heat drying
Temperature is 65-110 DEG C.
Described citrate buffer solution, concentration be 50mmol/L, pH be 4.8.
It is an advantage of the current invention that:
(1) ionic liquid pretreatment condition is relatively mild, requires low to experimental facilities.
(2) compared with untreated biomass material, after difunctionalization ionic liquid pretreatment, enzymolysis is imitated
Rate is remarkably reinforced, and reducing sugar yield improves.
(3) compared with traditional ionic liquid pretreatment technique, through difunctionalization ionic liquid pretreatment
After, cellulose and hemicellulose are farthest retained, for industrial common fermentation skill
For art, this process technique is most important.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is described further, but protection scope of the present invention is not subject to
The restriction of embodiment, if the present invention is made by the person skilled in the art in this field according to the invention described above content
Some nonessential improvement and adjustment, still fall within protection scope of the present invention.
Embodiment 1
[Et4N][Me2NC4SO3] 120 DEG C of pretreatment improve mallee bark enzymolysis efficiency
(1) pretreatment: precise 500mg by bark fines (particle diameter 40-80 mesh) and
10g[Et4N][Me2NC4SO3] ionic liquid, it is placed in 100ml round-bottomed flask, stirs at 120 DEG C
Mix 10 hours；After being cooled to room temperature, add 40ml deionized water wash, filtration, then use 40ml
Deionized water wash filtering residue 5 times, filtering residue remembers pretreated mallee bark after drying.
(2) enzymolysis experiment: accurate weighing pretreated mallee bark 20mg, is placed in 50ml tool plug conical flask,
Add 7ml citrate buffer (50mmol/L, pH 4.8) and cellulase, seal rearmounted
React in 160r/min, the constant temperature oscillator of 50 DEG C.Timing sampling 200ul, then takes out
Be placed in boiling water with quencher enzyme digestion reaction, centrifugal after, with high-performance liquid chromatogram determination glucose and
The concentration of xylose.According to the theoretical yield of glucose and xylose, Ji Keji in mallee bark before pretreatment
Final glucose and the yield of xylose, such as table 1.
Table 1
| Time (h) | 4 | 8 | 24 | 48 | 96 |
| Glucose yield (%) | 24.46 | 30.32 | 58.13 | 64.56 | 80.33 |
| Xylose yield (%) | 21.07 | 21.86 | 45.16 | 48.54 | 60.30 |
Embodiment 2
[Et4N][i-PrNHC3SO3] 120 DEG C of pretreatment improve mallee bark enzymolysis efficiency
(1) pretreatment: precise 500mg by bark fines (particle diameter 40-80 mesh) and
10g[Et4N][i-PrNHC3SO3] ionic liquid, it is placed in 100ml round-bottomed flask, stirs at 120 DEG C
Mix 10 hours；After being cooled to room temperature, add 40ml deionized water wash, filtration, then use 40ml
Deionized water wash filtering residue 5 times, filtering residue remembers pretreated mallee bark after drying.
(2) enzymolysis experiment: accurate weighing pretreated mallee bark 20mg, is placed in 50ml tool plug conical flask,
Adding 7ml citrate buffer (50mmol/L, pH 4.8) and cellulase, sealing is placed on
160r/min, reacts in the constant temperature oscillator of 50 DEG C.Timing sampling 200ul, then takes out rearmounted
With quencher enzyme digestion reaction in boiling water, after being centrifuged, with high-performance liquid chromatogram determination glucose and xylose
Concentration.According to the theoretical yield of glucose and xylose in mallee bark before pretreatment, can calculate final
Glucose and the yield of xylose, such as table 2.
Table 2
| Time (h) | 4 | 8 | 24 | 48 | 96 |
| Glucose yield (%) | 16.25 | 24.00 | 36.00 | 39.73 | 48.39 |
| Xylose yield (%) | 10.88 | 17.10 | 26.64 | 29.32 | 38.83 |
Embodiment 3
[Ch][Et2NC4SO3] 120 DEG C of pretreatment improve mallee bark enzymolysis efficiency
(1) pretreatment: precise 500mg mallee bark powder (particle diameter 40-80 mesh) and 10g [Ch] [Et2NC4SO3]
Ionic liquid, is placed in 100ml round-bottomed flask, stirs 10 hours at 120 DEG C；It is cooled to room temperature
After, add 40ml deionized water wash, filtration, then with 40ml deionized water wash filtering residue 5 times,
Filtering residue remembers pretreated mallee bark after drying.
(2) enzymolysis experiment: accurate weighing pretreated mallee bark 20mg, is placed in 50ml tool plug conical flask,
Adding 7ml citrate buffer (50mmol/L, pH 4.8) and cellulase, sealing is placed on
160r/min, reacts in the constant temperature oscillator of 50 DEG C.Timing sampling 200ul, then takes out rearmounted
With quencher enzyme digestion reaction in boiling water, after being centrifuged, with high-performance liquid chromatogram determination glucose and xylose
Concentration.According to the theoretical yield of glucose and xylose in mallee bark before pretreatment, can calculate final
Glucose and the yield of xylose, such as table 3.
Table 3
| Time (h) | 4 | 8 | 24 | 48 | 96 |
| Glucose yield (%) | 12.82 | 20.04 | 28.19 | 34.44 | 37.73 |
| Xylose yield (%) | 10.54 | 16.95 | 22.32 | 29.22 | 32.78 |
Embodiment 4
[Ch][n-BuNHC3SO3] 120 DEG C of pretreatment improve mallee bark enzymolysis efficiency
(1) pretreatment: precise 500mg mallee bark powder (particle diameter 40-80 mesh) and
10g[Ch][n-BuNHC3SO3] ionic liquid, it is placed in 100ml round-bottomed flask, stirs at 120 DEG C
Mix 10 hours；After being cooled to room temperature, add 40ml deionized water wash, filtration, then use 40ml
Deionized water wash filtering residue 5 times, filtering residue remembers pretreated mallee bark after drying.
(2) enzymolysis experiment: accurate weighing pretreated mallee bark 20mg, is placed in 50ml tool plug conical flask,
Add 7ml citrate buffer (50mmol/L, pH 4.8) and cellulase, seal rearmounted
React in 160r/min, the constant temperature oscillator of 50 DEG C.Timing sampling 200ul, then takes out
Be placed in boiling water with quencher enzyme digestion reaction, centrifugal after, with high-performance liquid chromatogram determination glucose and
The concentration of xylose.According to the theoretical yield of glucose and xylose, Ji Keji in mallee bark before pretreatment
Final glucose and the yield of xylose, such as table 4.
Table 4
| Time (h) | 4 | 8 | 24 | 48 | 96 |
| Glucose yield (%) | 11.69 | 14.96 | 20.39 | 22.43 | 23.37 |
| Xylose yield (%) | 8.48 | 10.49 | 17.78 | 19.52 | 21.32 |
Embodiment 5
[Et4N][Me2NC4SO3] 180 DEG C of pretreatment improve mallee bark enzymolysis efficiency
(1) pretreatment: precise 500mg mallee bark powder (particle diameter 40-80 mesh) and
10g[Et4N][Me2NC4SO3] ionic liquid, it is placed in 100ml round-bottomed flask, stirs at 180 DEG C
Mix 6 hours；After being cooled to room temperature, add 40ml deionized water wash, filtration, then use 40ml
Deionized water wash filtering residue 5 times, filtering residue remembers pretreated mallee bark after drying.
(2) enzymolysis experiment: accurate weighing pretreated mallee bark 20mg, is placed in 50ml tool plug conical flask,
Adding 7ml citrate buffer (50mmol/L, pH 4.8) and cellulase, sealing is placed on
160r/min, reacts in the constant temperature oscillator of 50 DEG C.96 hours sampling 200ul, then take out
It is placed in boiling water with quencher enzyme digestion reaction, after being centrifuged, with high-performance liquid chromatogram determination glucose and wood
The concentration of sugar.According to the theoretical yield of glucose and xylose in mallee bark before pretreatment, can calculate
Whole glucose and the yield of xylose, such as table 5.
Table 5
| Time (h) | 96 |
| Glucose yield (%) | 92.10 |
| Xylose yield (%) | 80.20 |
Embodiment 6
[Et4N][Me2NC4SO3] 140 DEG C of pretreatment improve Fast growth poplar enzymolysis efficiency
(1) pretreatment: precise 1000mg Fast growth poplar powder (particle diameter 40-80 mesh) and
10g[Et4N][Me2NC4SO3] ionic liquid, it is placed in 100ml round-bottomed flask, stirs at 140 DEG C
Mix 6 hours；After being cooled to room temperature, add 80ml deionized water wash, filtration, then use 80ml
Deionized water wash filtering residue 5 times, filtering residue remembers pretreated Fast growth poplar after drying.
(2) enzymolysis experiment: accurate weighing pretreated Fast growth poplar 20mg, is placed in 50ml tool plug conical flask,
Adding 7ml citrate buffer (50mmol/L, pH 4.8) and cellulase, sealing is placed on
160r/min, reacts in the constant temperature oscillator of 50 DEG C.96 hours sampling 200ul, then take out
It is placed in boiling water with quencher enzyme digestion reaction, after being centrifuged, with high-performance liquid chromatogram determination glucose and wood
The concentration of sugar.According to the theoretical yield of glucose and xylose in Fast growth poplar before pretreatment, can calculate
Whole glucose and the yield of xylose, such as table 6.
Table 6
| Time (h) | 96 |
| Glucose yield (%) | 85.20 |
| Xylose yield (%) | 63.20 |
Embodiment 7
[Emim][Me2NC4SO3] 150 DEG C of pretreatment improve Kenaf Core enzymolysis efficiency
(1) pretreatment: precise 2000mg Kenaf Core powder (particle diameter 40-80 mesh) and
10g[Emim][Me2NC4SO3] ionic liquid, it is placed in 100ml round-bottomed flask, stirs at 150 DEG C
Mix 6 hours；After being cooled to room temperature, add 120ml deionized water wash, filtration, then use 120ml
Deionized water wash filtering residue 5 times, filtering residue remembers pretreated Fast growth poplar after drying.
(2) enzymolysis experiment: accurate weighing pretreated Kenaf Core 20mg, is placed in 50ml tool plug conical flask,
Adding 7ml citrate buffer (50mmol/L, pH 4.8) and cellulase, sealing is placed on
160r/min, reacts in the constant temperature oscillator of 50 DEG C.96 hours sampling 200ul, then take out
It is placed in boiling water with quencher enzyme digestion reaction, after being centrifuged, with high-performance liquid chromatogram determination glucose and wood
The concentration of sugar.According to the theoretical yield of glucose and xylose in Kenaf Core before pretreatment, can calculate
Whole glucose and the yield of xylose, such as table 7.
Table 7
| Time (h) | 96 |
| Glucose yield (%) | 73.20 |
| Xylose yield (%) | 60.20 |
Comparative example 1
The most pretreated mallee bark enzymolysis
Accurate weighing the most pretreated mallee bark 20mg, is placed in 50ml tool plug conical flask, adds 7ml
Citrate buffer (50mmol/L, pH 4.8) and cellulase, seal and be placed on 160r/min, and 50
DEG C constant temperature oscillator in react.Timing sampling 200ul, then takes out and is placed in boiling water with quencher enzymolysis
Reaction, after being centrifuged, by the concentration of high-performance liquid chromatogram determination glucose and xylose.According to Fructus Vitis viniferae in mallee bark
Sugar and the theoretical yield of xylose, can calculate the yield of final glucose and xylose, such as table 8.
Table 8
| Time (h) | 4 | 8 | 24 | 48 | 96 |
| Glucose yield (%) | 2.95 | 3.86 | 6.48 | 7.14 | 8.83 |
| Xylose yield (%) | 7.28 | 8.21 | 15.11 | 16.91 | 18.20 |
Comparative example 2
The most pretreated Fast growth poplar enzymolysis
Accurate weighing the most pretreated Fast growth poplar powder 20mg, is placed in 50ml tool plug conical flask, adds
7ml citrate buffer (50mmol/L, pH 4.8) and cellulase, seal and be placed on 160r/min,
The constant temperature oscillator of 50 DEG C reacts.96 hours sampling 200ul, then take out and are placed in boiling water with quencher
Enzyme digestion reaction, after being centrifuged, by the concentration of high-performance liquid chromatogram determination glucose and xylose.According in Fast growth poplar
The theoretical yield of glucose and xylose, can calculate the yield of final glucose and xylose, such as table 9.
Table 9
| Time (h) | 96 |
| Glucose yield (%) | 6.23 |
| Xylose yield (%) | 15.20 |
Comparative example 3
The most pretreated Kenaf Core enzymolysis
Accurate weighing the most pretreated Kenaf Core powder 20mg, is placed in 50ml tool plug conical flask, adds
7ml citrate buffer (50mmol/L, pH 4.8) and cellulase, seal and be placed on 160r/min,
The constant temperature oscillator of 50 DEG C reacts.96 hours sampling 200ul, then take out and are placed in boiling water with quencher
Enzyme digestion reaction, after being centrifuged, by the concentration of high-performance liquid chromatogram determination glucose and xylose.According in Kenaf Core
The theoretical yield of glucose and xylose, can calculate the yield of final glucose and xylose, such as table 10.
Table 10
| Time (h) | 96 |
| Glucose yield (%) | 9.10 |
| Xylose yield (%) | 17.65 |
Claims (14)
1. the method that difunctionalization ionic liquid pretreatment biomass improve enzymolysis efficiency, it is characterised in that the party
Method follows the steps below:
(1) protista matter drying, pulverizing, its mean diameter controls at 20-200 mesh；
(2) with difunctionalization ionic liquid as pre-treatment solvents, according to the mass ratio of biomass and ionic liquid it is
1:2.5～25 mixing biomass and ionic liquid, stir 0.5～36h at room temperature～200 DEG C, be subsequently cooled to
Room temperature, adds the water of 2～10 times of ionic liquid volumes, filters, and washs filtering residue, obtains pretreatment after drying
After biomass；
(3) it is 1～10mg/ml to weigh pretreated biomass material and join citrate according to solid-to-liquid ratio
In buffer, then add cellulase according to the ratio of 5～30FPU/g pretreated biomass,
Reacting at 100-300r/min, 40-60 DEG C, enzymolysis time is 0.5-100h, and glucose in solutions yield is remote
Much larger than glucose amount produced by undressed biomass material.
One difunctionalization ionic liquid pretreatment biomass the most according to claim 1 improve enzymolysis effect
The method of rate, it is characterised in that: described difunctionalization ionic liquid be containing sulfonic acid anion base but the most fluorine-containing
-CHxSO3 -Anion, wherein x=0,1 or 2；And sulfonic acid anion base is containing basic group.
A kind of difunctionalization ionic liquid pretreatment biomass improve enzymolysis efficiency
Method, it is characterised in that: described basic group is-NHx, wherein X=0,1 or 2.
One difunctionalization ionic liquid pretreatment biomass the most according to claim 2 improve enzymolysis effect
The method of rate, it is characterised in that containing the ionic liquid of sulfonic acid anion base: preferably containing sulfonic acid anion base
The anion structure of alkali ionic liquid is as follows:
M1And M2It is independently ,-H ,-CH3,-C2H5, C3-C16Straight chain or branched alkane, alkene
Hydrocarbon, aromatic hydrocarbons, monosubstituted aromatic hydrocarbons, di-substituted aryl hydrocarbon and polysubstituted aromatic hydrocarbons；N is 1 or 2；X is-H ,-CH3、
-C2H5, C3-C8Straight chain or branched alkane, alkene, aromatic hydrocarbons, monosubstituted aromatic hydrocarbons, di-substituted aryl hydrocarbon and
Polysubstituted aromatic hydrocarbons.
One difunctionalization ionic liquid pretreatment biomass the most according to claim 2 improve enzymolysis effect
The method of rate, it is characterised in that containing the ionic liquid of sulfonic acid anion base: described anion is preferably
[Et2NC3SO3],[Et2NC4SO3],[Me2NC4SO3],[Me2NC3SO3],
[n-BuNHC3SO3],[n-BuNHC4SO3],[n-HeNHC3SO3],[i-PrNHC3SO3],[i-PrNHC4SO3],[i
-BuNHC3SO3],[i-BuNHC4SO3],[MeNHC4SO3],[MeNHC3SO3],[EtNHC4SO3] or
[EtNHC3SO3]。
One difunctionalization ionic liquid pretreatment biomass the most according to claim 2 improve enzymolysis effect
The method of rate, it is characterised in that containing the ionic liquid of sulfonic acid anion base: described containing sulfonic acid anion base
The cation of ionic liquid be imidazoles organic cation, pyridines organic cation, the organic sun of piperidines from
Son, pyroles organic cation, quaternary ammonium salt organic cation, quaternary phosphonium salt organic cation or choline
Organic cation.
7. the side of enzymolysis efficiency is improved according to claim 6 difunctionalization ionic liquid pretreatment biomass
Method, it is characterised in that:
Imidazoles organic cation general structure is:
Pyridines organic cation general structure is:
Piperidines organic cation general structure is:
Pyroles organic cation general structure is:
Quaternary ammonium salt organic cation general structure is:
Quaternary phosphonium salt organic cation general structure is:
Choline organic cation structure is:
Wherein, R1-R6It is independently ,-CH3,-C2H5Or C3-C6Straight chain or branched alkane
Or alkene；R7-R10It is independently ,-CH3,-C2H5Or C3-C15Straight chain or branched alkane or
Alkene.
One difunctionalization ionic liquid pretreatment biomass the most according to claim 2 improve enzymolysis effect
The method of rate, it is characterised in that containing the ionic liquid of sulfonic acid anion base: cation is preferably tetramethyl amine
(Me4N), tetraethyl amine (Et4N), 4-butyl amine (Bu4N), 1-methyl-3-ethyl imidazol(e) (Emim),
Choline (Ch).
A kind of difunctionalization ionic liquid pretreatment biomass improve enzymolysis efficiency
Method, described difunctionalization ionic liquid, it is characterised in that: preferably ionic liquid is combined as:
Cation is tetramethyl amine (Me4N), tetraethyl amine (Et4N), 4-butyl amine (Bu4N), 1-methyl
-3-ethyl imidazol(e) (Emim), the one in choline (Ch)；
Anion is [Et2NC3SO3],[Et2NC4SO3],[Me2NC4SO3],[Me2NC3SO3],
[n-BuNHC3SO3],[n-BuNHC4SO3],[n-HeNHC3SO3],[i-PrNHC3SO3],[i-PrNHC4SO3],[i
-BuNHC3SO3],[i-BuNHC4SO3],[MeNHC4SO3],[MeNHC3SO3],[EtNHC4SO3],
[EtNHC3SO3One in].
One difunctionalization ionic liquid pretreatment biomass the most according to claim 9 improve enzymolysis
The method of efficiency, it is characterised in that described difunctionalization ionic liquid: preferably ionic liquid is [Ch] [Et2NC3SO3],[Ch][Me2NC4SO3],[Ch][n-BuNHC3SO3],[Ch][n-HeNHC3SO3],[Ch][i-PrN
HC3SO3],[Ch][i-PrNHC4SO3],[Ch][i-BuNHC3SO3],[Et4N][i-PrNHC3SO3],[Et4N][Me2
NC4SO3],[Et4N][Et2NC3SO3],[Et4N][n-BuNHC3SO3],[Et4N][i-BuNHC3SO3],[Me4
N][MeNHC3SO3], [Me4N][MeNHC4SO3], [Emim] [MeNHC3SO3] one of which or several
The mixture planted.
11. one difunctionalization ionic liquid pretreatment biomass according to claim 1 improve enzymolysis effect
The method of rate, it is characterised in that: described protista raw material has various trees, agricultural crop straw, agriculture
One or several mixture in Product processing industry side-product, feces of livestock and poultry and energy crop etc., described tree
Wood is cork and/or hardwood.
12. improve enzymolysis effect according to the one difunctionalization ionic liquid pretreatment biomass described in claim 1
The method of rate, it is characterised in that: described protista raw material preferable particle size is: 40-80 mesh；Described
Pretreatment preferable temperature is: 50-180 DEG C.
13. one difunctionalization ionic liquid pretreatment biomass according to claim 1 improve enzymolysis
The method of efficiency, it is characterised in that: after described pretreatment protista matter drying means be lyophilization or
Person's heat drying；Heat drying temperature is 65-110 DEG C.
14. improve enzymolysis according to the one difunctionalization ionic liquid pretreatment biomass described in claim 1
The method of efficiency, it is characterised in that: described citrate buffer solution, concentration be 50mmol/L, pH be 4.8.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101589153A (en) * | 2007-01-23 | 2009-11-25 | 巴斯夫欧洲公司 | Method for producing glucose by enzymatic hydrolysis of cellulose that can be pretreated with an ionic liquid containing a polyatomic anion |
| WO2014140643A1 (en) * | 2013-03-15 | 2014-09-18 | Imperial Innovations Limited | Treatment |
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2015
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101589153A (en) * | 2007-01-23 | 2009-11-25 | 巴斯夫欧洲公司 | Method for producing glucose by enzymatic hydrolysis of cellulose that can be pretreated with an ionic liquid containing a polyatomic anion |
| WO2014140643A1 (en) * | 2013-03-15 | 2014-09-18 | Imperial Innovations Limited | Treatment |
Non-Patent Citations (1)
| Title |
|---|
| PEIFANG YAN等: ""Factionation of lignin from eucalyptus bark using amine-sufonate functionalized ionic liquids"", 《GREEN CHEMISTRY》 * |
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