CN106591395A - Method for improving lignocellulose enzymolysis efficiency by lignocellulose pretreatment through ultrasonic-microwave synchronously-assisted ionic liquid system - Google Patents
Method for improving lignocellulose enzymolysis efficiency by lignocellulose pretreatment through ultrasonic-microwave synchronously-assisted ionic liquid system Download PDFInfo
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Abstract
The invention belongs to the technical field of resourceful utilization of lignocellulose biomass, aims to provide an environment-friendly efficient lignocellulose pretreatment method and relates to a novel method for improving lignocellulose enzymolysis efficiency by extracting lignin components in lignocelluloses through an ionic liquid-water mixed system assisted by synergistic effect of ultrasonic-microwave energy. Compared with a traditional ionic liquid pretreatment method, the method has the advantages that synergistic ultrasonic-microwave energy is initiatively used for increasing the ratio that the ionic liquid-water mixed system extracts the lignin, enhancing accessibility of cellulase and the cellulose components in the lignocelluloses and improving the lignocellulose enzymolysis efficiency; ionic liquid consumption during pretreatment is reduced substantially through mixing of water, so that technological costs are saved; the method is short in treatment time, mild and green in conditions and simple in technology.
Description
Technical field
The present invention relates to a kind of ultrasonic-microwave synchronously assists the mixed system pretreatment wood fibre of ionic liquid-water
The method that element improves its enzymolysis efficiency, belongs to the application technology as the second resource field of lignocellulose-like biomass.
Background technology
Lignocellulose is the huge biomass resource of reserves on the earth, and its main source has agricultural crop straw, trees, energy
Source plant and weeds etc..With a series of emerging in large numbers for environment and energy problem, reproducible lignocellulose has become the mankind
Focus development and the energy resources for utilizing.Because its energy utilization rate is low and the problems such as serious environmental pollution, directly burns wooden
The tradition of cellulose utilizes form, is gradually substituted by other approach；At present, lignocellulose-like biomass is passed through into bioanalysises
Hydrolysis generates reducing sugar, then Jing fermentables obtain the study hotspot of fuel and other chemicals Cheng Xin.
Enzyme hydrolysiss lignocellulose prepares reducing sugar technique because of environmental friendliness, mild condition and reducing sugar high income again
Favored, but the mineral crystal structure of lignocellulose inner tight but hinders accessibility of the enzyme molecule to substrate, significantly
Limit enzymolysis efficiency.Lignocellulose is mainly made up of cellulose, hemicellulose and lignin；Wherein cellulose is by β-D-
Glucose is formed by connecting by β-Isosorbide-5-Nitrae glycosidic bond, is chain heteromorphic polymer；The change that hemicellulose is made up of various sugar units
Learn the non-homogeneous polysaccharide of structure；Lignin be by benzene oxide unit with C-C keys and C-O-C ehter bonds be formed by connecting it is unformed
Complicated netted heteropolymer；Wherein lignin is " viscose glue " that lignocellulose key component links together so as to indissoluble
Yu Shui, also links just because of lignin and hemicellulose and realizes the parcel to cellulose and winding effect so as to become enzyme
Solution and the significant obstacle of microbial degradation.
During biorefinery, in order to improve the efficiency that cellulase is hydrolyzed to lignocellulose, need to wooden fibre
Dimension element carries out pretreatment to break its compact texture, except constituents for suppressing such as delignifications, reduces the degree of crystallinity of cellulose, maximum limit
Improve the contacting efficiency of enzyme and cellulose in degree ground.Preprocess method traditional at present includes Physical, chemical method, physical-chemical process
And bioanalysises, but majority to there is environmental pollution big, the shortcomings of low production efficiency.As a kind of novel green solvent, ionic liquid
It is the salt being made up of organic cation and organic or inorganic anion.Ionic liquid has good solubility to lignocellulose component
Can, and with non-volatile, environmental pollution is little, the advantages of reusable, at present at the pre- place of lignocellulose-like biomass
It is used widely in reason.The pretreating process mainly uses the superpower solvability of ionic liquid, first by lignocellulose
In cellulose dissolution, be subsequently adding other solvents（Anti-solvent）Separate out Cellulose precipitates（Regeneration）, to destroy its crystal knot
Structure.However, because viscosity of il is big, the mass transfer in preprocessing process can be affected, increase operation difficulty；While high cost
It is also problem present in ionic liquid pretreatment.At present existing solution is to add water in ionic liquid to reduce gluing
Degree, strengthens mass transfer, and reducing the presence of the water in cost, but ionic liquid can seriously hinder the dissolving of cellulose again, therefore the dissolving
Technique requires water content very harsh（1% need to be generally less than）, this just must carry out long-time high temperature to raw material and ionic liquid
Processed, causes energy consumption excessive, seriously reduces the economic feasibility of the pretreating process.
The content of the invention
The technical problem to be solved is：Lignocellulose is carried out in preprocessing process using ionic liquid,
Need the water content control in ionic liquid in very low range, to promote cellulose dissolution in ionic liquid, during deposit
Carrying out long-time high temperature dehydration process to raw material and ionic liquid, energy consumption is big, high cost the problems such as.
The present invention employs a kind of and diverse technology design of prior art, i.e., do not consider ionic liquid to fiber
The deliquescent problem of element, and directly adopt and lignin is divided lignin from cellulose with the ionic liquid compared with highly dissoluble
Separate out, meanwhile, viscosity of il, reinforcing mass transfer, reduces cost are reduced using more water is added, in this process,
It is aided with the superposition of two kinds of energy of ultrasonic-microwave, effectively removes the lignin in lignocellulose, being capable of reinforcing fiber element enzyme pair
The hydrolysis efficiency of cellulose, significantly improves fermentable reducing sugar（Glucose and xylose）Yield；And after the pre-treatment, also together
When avoid the problem for needing again to extract the cellulose in ionic liquid with anti-solvent again in routine techniquess, directly will process
Solidss afterwards send into enzymolysis operation.
Specifically technical scheme is：
A kind of method that ultrasonic-microwave synchronization assisting ion liquid system preprocessing lignocellulose improves its enzymolysis efficiency,
Comprise the steps：
1st step, lignocellulose raw material is added into the mixed solution of ionic liquid and water, in additional microwave/ultrasonic field
Under effect, heat treated is carried out；After solidss are leached, through washing, drying, pretreated lignocellulose is obtained；
2nd step, through enzymolysis processing, it is main to obtain with reducing sugar glucose and xylose to the lignocellulose that the 1st step is obtained
The saccharified liquid of composition.
In one embodiment, in the 1st described step, the ionic liquid of employing refers to there is lignin compared with good dissolving
Property, there is relatively low deliquescent ionic liquid to cellulose.
In one embodiment, described ionic liquid refers to [choline] [ornithine] [Cho] [Orn], and it has such as
Lower shown structure：
In one embodiment, in the mixed solution of ionic liquid and water, mass percent scope is 25 shared by ionic liquid
～99.9wt%, more preferably 45～55wt%.
In one embodiment, lignocellulose raw material is agricultural crop straw, forest, bamboo and wood, wild draft, the energy
Crop or their mixture, its raw material particle size scope is 0.1～5cm.
In one embodiment, lignocellulose raw material and mixed solution mass ratio are 1:5～1:25.
In one embodiment, 15～1000W of microwave power, 1500～3000MHz of microwave frequency.
In one embodiment, 200～1000W of ultrasonic power, 15～30KHz of ultrasonic frequency.
In one embodiment, enzymolysis is that lignocellulose after the pre-treatment adds appropriate cellulase to carry out enzyme
Solution saccharification react.
In one embodiment, enzymolysis process be by pretreated lignocellulose according to solid-to-liquid ratio be 1～10
Mg/mL mixes with citrate buffer, adds cellulase, in 150～250r/min speeds of agitator, 35～65 DEG C of temperature
Under degree, react 1～72 hour.
In one embodiment, cellulose enzyme amount is added to be 5～1000U/g substrates in feed liquid.
In one embodiment, the cellulase is commercially available extensive stock enzyme preparation or microorganism direct fermentation
The enzyme liquid for obtaining.
Beneficial effect
Present invention firstly provides a kind of new method of ionic liquid-water mixed system preprocessing lignocellulose, existing same
Class pretreatment system is usually the pure ionic liquid having to cellulose compared with high-solvency, such as imidazolium ionic liquid.
The present invention need not investigate solvability of the ionic liquid to cellulose, only be absorbed in the leaching effect of its lignin, be firstly introduced
The mixed system of [choline] [ornithine]-water is as preprocessing solution and preferably while relatively low using having to cellulose
The ionic liquid of solvability, it is to avoid the loss of cellulose, while synchronously there is system as assistant strip by the use of ultrasonic-microwave
Part, the problems such as not only solve slow mass transfer caused by pure viscosity of il is high in preprocessing process and big operation difficulty, and mixes
The key that links of cellulose and lignin has effectively been broken in system pretreatment, removes lignin or hemicellulose, makes cellulose sudden and violent
Expose, increase accessibility of the cellulase to substrate, it will be apparent that improve the enzymolysis efficiency of lignocellulose, reduce ion
The consumption of liquid, economize on resources reduces cost；Meanwhile, the characteristics of the ionic liquid for being adopted has environmentally friendly.
The present invention strengthens first the effect of solution process lignocellulose using ultrasonic-microwave collaboration energy field simultaneously
Really.The invention has the advantages that：In ultrasound wave and the work station of the noiseless combination of microwave, the collaboration of two kinds of energy is processed
Sonic oscillation and ripple cavitation effect is reached and is preferably combined, ionic liquid can be strengthened to the moistening of lignocellulose and molten
Swollen effect, makes loose, the mechanical strength decrease that lignocellulosic structure becomes, and improves dissolution of the ionic liquid to lignin and separates
Effect, while ionic liquid solution can be entered preferably inside lignocellulose, breaks lignin and hemicellulose to cellulose
Constraint, cellulase and fiber have bigger contact area, solids separation is being gone out and to be applied to follow-up enzymolysis anti-
Ying Zhong, can promote enzyme hydrolysiss to carry out, and obtain higher reducing sugar yield.Ultrasonic-microwave collaboration preconditioning technique can be saved
Save time, without the need for high temperature, mild condition is simple to operate, and environmental protection, low cost, subsequent treatment is simple.
Specific embodiment
Embodiment of the present invention and produced pretreating effect, but this are further illustrated by embodiment and comparison example
The protection domain of invention is not limited to the content listed by embodiment.Following embodiments are the more excellent embodiment of the present invention, but are sent out
Bright embodiment is not limited by above-mentioned fact Example, and other any spirit without departing from the present invention are lower with principle to be made
Change, modification, replacement, combination, simplify, should be equivalent substitute mode, be included within protection scope of the present invention.
Embodiment 1：Utilize [Cho] [Orn] to rice straw pretreatment to lift its enzymolysis efficiency.
A) pretreatment：The g rice straw powder of accurate weighing 0.25（150～350 μm）With 4 g [Cho] [Orn],
In being collectively disposed at 20 mL triangular flasks, stir 12 hours at 95 DEG C；Room temperature is then cooled to, 0.5 mL supernatant is taken, is utilized
Determined by ultraviolet spectrophotometry lignin extraction rate.
B) sample is subsequently prepared：The NaOH solution of 1 times of ionic liquid volume is slowly added to in pretreatment system（0.1
mol/L）Dilution, filter, then with the deionized water wash filtering residue 4 times of 2 times of volumes, filtering residue Jing 60 DEG C dryings colourless to supernatant
Pretreated rice straw sample is obtained afterwards.
C) digest：The above-mentioned g of pretreated rice straw 0. 05 of accurate weighing, in being placed in the triangular flask of 50 mL, plus
Enter 20 mL citrate buffers (50 mmol/L, pH 4.8) and 45 U commercial fibers element enzyme, 200 are placed in after sealing
React in r/min, 50 DEG C of constant temperature oscillator.The μ l of timing sampling 200, are centrifuged after boiling 5 minutes（10000 g）, using efficient
Liquid chromatogram measuring glucose and xylose concentration, until glucose and xylose concentration reach maximum, terminating reaction in hydrolyzed solution.Root
Before Data preprocess in rice straw glucose and xylose theoretical yield.
Embodiment 2：[Cho] [Orn]-water mixed system is utilized to lift its enzymolysis efficiency to rice straw pretreatment.
A) pretreatment：Accurate weighing 0.25g rice straw powder（150～350 μm）, 2 g [Cho] [Orn] ion
Liquid and 2 g deionized waters, in being collectively disposed at 20 mL conical flask with stopper, stir 12 hours at 95 DEG C, are then cooled to room
Temperature；0.5ml supernatant is taken using determined by ultraviolet spectrophotometry lignin extraction rate.
B) sample is subsequently prepared：The NaOH solution of 1 times of ionic liquid volume is slowly added to in pretreatment system（0.1
mol/L）Dilution, filter, then with the deionized water wash filtering residue 4 times of 2 times of volumes, filtering residue Jing 60 DEG C dryings colourless to supernatant
Pretreated rice straw sample is obtained afterwards.
C) digest：The above-mentioned pretreated rice straw 0.05g of accurate weighing, is placed in the conical flask with cover of 50 mL
In, 20 mL citrate buffers (50 mmol/L, pH 4.8) and 45 U commercial fibers element enzyme are added, it is placed in after sealing
React in 200 r/min, 50 DEG C of constant temperature oscillator.In 0 h, 12 h, the μ L of 24 h timing samplings 200, after boiling 5 minutes from
The heart（10000 g）, using high-performance liquid chromatogram determination glucose and xylose concentration, until glucose and xylose concentration in hydrolyzed solution
Reach maximum, terminating reaction.According to the theoretical yield of glucose and xylose in rice straw before pretreatment.
Embodiment 3：Ultrasonic-microwave assists [Cho] Orn- water mixed systems（50%：50%）Preprocessing straw stalk improves it
The method of enzymolysis efficiency.
A) pretreatment：The g rice straw powder of accurate weighing 0.25（150 ~ 350μm）, 2 g [Cho] [Orn] from
Sub- liquid and 2 g deionized waters, during 20 mL triangular flasks are put in jointly, are placed in ultrasonic-microwave concerted reaction device（Nanjing elder generation Larus ridibunduss
Instrument manufacturing company limited）In；Setting condition is：Supersonic frequency 25KHZ, power 800W;The MHZ of microwave frequency 2450, work(
Rate 1000W, is kept for 6 hours under the conditions of 95 DEG C.Room temperature is then cooled to, 0.5ml supernatant is taken and is surveyed using ultraviolet spectrophotometry
Determine lignin extraction rate.
B) sample is subsequently prepared:The NaOH solution of 1 times of ionic liquid volume is slowly added to in pretreatment system
（0.1 mol/L）Dilution, filter, then with the deionized water wash filtering residue 4 times of 2 times of volumes, filtering residue Jing 60 colourless to supernatant
Pretreated rice straw sample is obtained after DEG C drying.
C) digest：The above-mentioned g of pretreated rice straw 0.05 of accurate weighing, in being placed in the triangular flask of 50 mL, plus
Enter 20 mL citrate buffers (50 mmol/L, pH 4.8) and 45 U commercial fibers element enzyme, 200 are placed in after sealing
React in r/min, 50 DEG C of constant temperature oscillator.The μ L of timing sampling 200, are centrifuged after boiling 5 minutes（10000 g）, using efficient
Liquid chromatogram measuring glucose and xylose concentration, until glucose and xylose concentration reach maximum, terminating reaction in hydrolyzed solution.Root
Before Data preprocess in rice straw glucose and xylose theoretical yield.
Embodiment 4：Ultrasonic-microwave assists [Cho] Orn- water mixed systems（30%：70%）Preprocessing straw stalk improves it
The method of enzymolysis efficiency.
A) pretreatment：The g rice straw powder of accurate weighing 0.25（150 ~ 350μm）, 1.2 g [Cho] [Orn]
Ionic liquid and 2.8 g deionized waters, during 20 mL triangular flasks are put in jointly, are placed in ultrasonic-microwave concerted reaction device（Nanjing
Xian Ou instrument manufacturings company limited）In；Setting condition is：Supersonic frequency 25KHZ, power 800W;Microwave frequency 2450
MHZ, power 1000W, are kept for 6 hours under the conditions of 95 DEG C.Room temperature is then cooled to, 0.5ml supernatant is taken and is utilized ultraviolet spectrometry
Spectrphotometric method for measuring lignin extraction rate.
B) sample is subsequently prepared:The NaOH solution of 1 times of ionic liquid volume is slowly added to in pretreatment system
（0.1 mol/L）Dilution, filter, then with the deionized water wash filtering residue 4 times of 2 times of volumes, filtering residue Jing 60 colourless to supernatant
Pretreated rice straw sample is obtained after DEG C drying.
C) digest：The above-mentioned g of pretreated rice straw 0.05 of accurate weighing, in being placed in the triangular flask of 50 mL, plus
Enter 20 mL citrate buffers (50 mmol/L, pH 4.8) and 45 U commercial fibers element enzyme, 200 are placed in after sealing
React in r/min, 50 DEG C of constant temperature oscillator.The μ L of timing sampling 200, are centrifuged after boiling 5 minutes（10000 g）, using efficient
Liquid chromatogram measuring glucose and xylose concentration, until glucose and xylose concentration reach maximum, terminating reaction in hydrolyzed solution.Root
Before Data preprocess in rice straw glucose and xylose theoretical yield.
Embodiment 5：Ultrasonic-microwave assists [Cho] Orn- water mixed systems（70%：30%）Preprocessing straw stalk improves it
The method of enzymolysis efficiency.
A) pretreatment：The g rice straw powder of accurate weighing 0.25（150 ~ 350μm）, 2.8 g [Cho] [Orn]
Ionic liquid and 1.2 g deionized waters, during 20 mL triangular flasks are put in jointly, are placed in ultrasonic-microwave concerted reaction device（Nanjing
Xian Ou instrument manufacturings company limited）In；Setting condition is：Supersonic frequency 25KHZ, power 800W;Microwave frequency 2450
MHZ, power 1000W, are kept for 6 hours under the conditions of 95 DEG C.Room temperature is then cooled to, 0.5ml supernatant is taken and is utilized ultraviolet spectrometry
Spectrphotometric method for measuring lignin extraction rate.
B) sample is subsequently prepared:The NaOH solution of 1 times of ionic liquid volume is slowly added to in pretreatment system
（0.1 mol/L）Dilution, filter, then with the deionized water wash filtering residue 4 times of 2 times of volumes, filtering residue Jing 60 colourless to supernatant
Pretreated rice straw sample is obtained after DEG C drying.
C) digest：The above-mentioned g of pretreated rice straw 0.05 of accurate weighing, in being placed in the triangular flask of 50 mL, plus
Enter 20 mL citrate buffers (50 mmol/L, pH 4.8) and 45 U commercial fibers element enzyme, 200 are placed in after sealing
React in r/min, 50 DEG C of constant temperature oscillator.The μ L of timing sampling 200, are centrifuged after boiling 5 minutes（10000 g）, using efficient
Liquid chromatogram measuring glucose and xylose concentration, until glucose and xylose concentration reach maximum, terminating reaction in hydrolyzed solution.Root
Before Data preprocess in rice straw glucose and xylose theoretical yield.
Comparative example 1：Ultrasound wave individually assists [Cho] [Orn]-water mixed system preprocessing straw.
A) pretreatment：The g rice straw powder of accurate weighing 0.25（150 ~ 350μm）, 2 g [Cho] [Orn] from
Sub- liquid and 2 g deionized waters, in being collectively disposed at 20 mL triangular flasks, in being placed in ultrasonic-microwave concerted reaction device；Bar is set
Part is：Supersonic frequency 25KHZ, power 800W;The MHZ of microwave frequency 0, the W of power 0, stir 6 hours at 95 DEG C；Subsequently
Room temperature is cooled to, room temperature is then cooled to；0.5ml supernatant is taken using determined by ultraviolet spectrophotometry lignin extraction rate.
B) sample is subsequently prepared：The NaOH solution of 1 times of ionic liquid volume is slowly added to in pretreatment system（0.1
mol/L）Dilution, filter, then with the deionized water wash filtering residue 4 times of 2 times of volumes, filtering residue Jing 60 DEG C dryings colourless to supernatant
Pretreated rice straw sample is obtained afterwards.
B) digest：The above-mentioned g of pretreated rice straw 0.05 of accurate weighing, in being placed in the triangular flask of 50 mL, plus
Enter 20 mL citrate buffers (50 mmol/L, pH 4.8) and 45 U commercial fibers element enzyme, 200 are placed in after sealing
React in r/min, 50 DEG C of constant temperature oscillator.The μ L of timing sampling 200, are centrifuged after boiling 5 minutes（10000 g）, using height
Effect liquid phase chromatogram determines glucose and xylose concentration, until glucose and xylose concentration reach maximum, terminating reaction in hydrolyzed solution.
According to the theoretical yield of glucose and xylose in rice straw before pretreatment.
Comparative example 2：Microwave assistant [Cho] [Orn]-water mixed system preprocessing straw improves its enzymolysis efficiency.
A) pretreatment：The g rice straw powder of accurate weighing 0.25（150 ~ 350 μm）, 2 g [Cho] [Orn] from
Sub- liquid and 2 g deionized waters, during 20 mL triangular flasks are put in jointly, in being placed in ultrasonic-microwave concerted reaction device；Bar is set
Part is：The KHZ of supersonic frequency 0, the W of power 0;The MHZ of microwave frequency 2450, power 1000W, holding 6 is little under the conditions of 95 DEG C
When.Room temperature is then cooled to, 0.5 mL supernatant is taken using determined by ultraviolet spectrophotometry lignin extraction rate.
B) sample is subsequently prepared：The NaOH solution of 1 times of ionic liquid volume is slowly added to in pretreatment system（0.1
mol/L）Dilution, filter, then with the deionized water wash filtering residue 4 times of 2 times of volumes, filtering residue Jing 60 DEG C dryings colourless to supernatant
Pretreated rice straw sample is obtained afterwards.
C) digest：The above-mentioned g of pretreated rice straw 0.05 of accurate weighing, in being placed in the triangular flask of 50 mL, plus
Enter 20 mL citrate buffers (50 mmol/L, pH 4.8) and 45U commercial fibers element enzyme, 200 are placed in after sealing
React in r/min, 50 DEG C of constant temperature oscillator.The μ L of timing sampling 200, are centrifuged after boiling 5 minutes（10000 g）, using efficient
Liquid chromatogram measuring glucose and xylose concentration, until glucose and xylose concentration reach maximum, terminating reaction in hydrolyzed solution.Root
Before Data preprocess in rice straw glucose and xylose theoretical yield.
The ultrasonic-microwave of comparative example 3 assists ionic liquid-water mixed system preprocessing straw stalk to improve the side of its enzymolysis efficiency
Method.
A) pretreatment：The g rice straw powder of accurate weighing 0.25（150 ~ 350μm）, 2 g other species from
Sub- liquid（Respectively from 1- ethyl-3-methylimidazole villaumites, 1- pi-allyl -3- methylimidazole villaumites and 1- ethyl -3- methyl -
Imidazoles acetate, respectively as the 1st～3 group）With 2 g deionized waters, during 20 mL triangular flasks are put in jointly, be placed in ultrasound wave-
Microwave cooperating reactor（Nanjing Xian Ou instrument manufacturings company limited）In；Setting condition is：Supersonic frequency 25KHZ, power
800W;The MHZ of microwave frequency 2450, power 1000W, are kept for 6 hours under the conditions of 95 DEG C.It is then cooled to room temperature.It is subsequently cold
But to room temperature, 0.5 mL supernatant is taken using determined by ultraviolet spectrophotometry lignin extraction rate.
B) sample is subsequently prepared:The NaOH solution of 1 times of ionic liquid volume is slowly added to in pretreatment system
（0.1 mol/L）Dilution, filter, then with the deionized water wash filtering residue 4 times of 2 times of volumes, filtering residue Jing 60 colourless to supernatant
Pretreated rice straw sample is obtained after DEG C drying.
C) digest：The above-mentioned g of pretreated rice straw 0.05 of accurate weighing, in being placed in the triangular flask of 50 mL, plus
Enter 20 mL citrate buffers (50 mmol/L, pH 4.8) and 45 U commercial fibers element enzyme, 200 are placed in after sealing
React in r/min, 50 DEG C of constant temperature oscillator.The μ L of timing sampling 200, are centrifuged after boiling 5 minutes（10000 g）, using efficient
Liquid chromatogram measuring glucose and xylose concentration, until glucose and xylose concentration reach maximum, terminating reaction in hydrolyzed solution.Root
Before Data preprocess in rice straw glucose and xylose theoretical yield.
Impact and corresponding enzymatic saccharification efficiency of the different pretreatments condition of table 1 to the lignin extraction rate of rice straw.
As shown in table 1, by contrasting with embodiment 1, glued for reducing ionic liquid by adding water in embodiment 2
Degree, but due in water the separating effect of lignin it is bad, had influence on separation of the lignin from cellulose, also result in
Follow-up enzyme digestion reaction effect is bad；Found out by embodiment 3, under conditions of the process of ultrasound-microwave cooperating, two kinds of reducing sugar
Final conversion ratio can exceed that embodiment 1, it was demonstrated that ultrasonic-microwave assists pretreatment to reduce ionic liquid usage amount
In the case of, promote the expansion of the internal structure of cellulose, lignin is preferably separated in system with cellulose, and energy
Significantly improve the conversion sugar yield of rice straw.Wherein can be seen that ultrasonic-microwave collaboration by data and process sample to wood
The leaching efficiency of quality is obviously improved effect, the reason for can lift saccharification efficiency so as to explain this method.In addition, logical
3～embodiment of embodiment 5 is crossed as can be seen that the amount for adding water controls when under the conditions of preferably, can effectively make lignin
Separated in mixed solution with cellulose, make hydrolysis result more preferable.
As shown in table 1, by further compared with comparative example 1 and comparative example 2, it was demonstrated that the ultrasound applied in embodiment 3
Ripple-microwave cooperating treatment conditions and simple ultrasonic Treatment or simple microwave treatment, have larger to improving sugar yield aspect
The raising of degree, this shows that the instantaneous synergism of two kinds of energy is located in advance to ionic liquid [Cho] [Orn]-water mixed system
Reason rice straw have be obviously improved effect.
By the parallel test of embodiment 3 and 3 groups of glyoxaline ion liquids of reference examples 3 can be seen that imidazoles from
Sub- liquid is larger for the dissolubility of cellulose, is not suitable for the technology design of the present invention, can cause cellulose in ionic liquid
In solution loss, decline final enzyme digestion reaction yield.
Claims (10)
1. a kind of ultrasonic-microwave synchronization assisting ion liquid system preprocessing lignocellulose improves the side of its enzymolysis efficiency
Method, it is characterised in that comprise the steps：
1st step, lignocellulose raw material is added into the mixed solution of ionic liquid and water, in additional microwave-ultrasonic field
Under effect, heat treated is carried out；After solidss are leached, through washing, drying, pretreated lignocellulose is obtained；
2nd step, through enzymolysis processing, it is main to obtain with reducing sugar glucose and xylose to the lignocellulose that the 1st step is obtained
The saccharified liquid of composition.
2. ultrasonic-microwave synchronization assisting ion liquid system preprocessing lignocellulose according to claim 1 is improved
The method of its enzymolysis efficiency, it is characterised in that in the 1st described step, the ionic liquid of employing refers to have preferably to lignin
Dissolubility, there is relatively low deliquescent ionic liquid to cellulose.
3. ultrasonic-microwave synchronization assisting ion liquid system preprocessing lignocellulose according to claim 1 is improved
The method of its enzymolysis efficiency, it is characterised in that described ionic liquid refers to [choline] [ornithine] [Cho] [Orn], and it has
Structure shown below：
。
4. ultrasonic-microwave synchronization assisting ion liquid system preprocessing lignocellulose according to claim 1 is improved
The method of its enzymolysis efficiency, it is characterised in that in the mixed solution of ionic liquid and water, mass percent model shared by ionic liquid
It is 25～99.9wt% to enclose, more preferably 45～55wt%.
5. ultrasonic-microwave synchronization assisting ion liquid system preprocessing lignocellulose according to claim 1 is improved
The method of its enzymolysis efficiency, it is characterised in that lignocellulose raw material be agricultural crop straw, forest, bamboo and wood, wild draft,
Energy crop or their mixture, its raw material particle size scope is 0.1～5cm.
6. ultrasonic-microwave synchronization assisting ion liquid system preprocessing lignocellulose according to claim 1 is improved
The method of its enzymolysis efficiency, it is characterised in that lignocellulose raw material is 1 with mixed solution mass ratio:5～1:25.
7. ultrasonic-microwave synchronization assisting ion liquid system preprocessing lignocellulose according to claim 1 is improved
The method of its enzymolysis efficiency, it is characterised in that 15～1000W of microwave power, 1500～3000MHz of microwave frequency；Ultrasound wave work(
200～1000W of rate, 15～30KHz of ultrasonic frequency.
8. ultrasonic-microwave synchronization assisting ion liquid system preprocessing lignocellulose according to claim 1 is improved
The method of its enzymolysis efficiency, it is characterised in that enzymolysis is that lignocellulose after the pre-treatment adds appropriate cellulase to enter
Row enzymatic saccharification reacts；It by pretreated lignocellulose according to solid-to-liquid ratio is 1～10 mg/mL and Fructus Citri Limoniae that enzymolysis process is
Phthalate buffer mixes, and adds cellulase, at a temperature of 150～250r/min speeds of agitator, 35～65 DEG C, reaction 1～
72 hours.
9. ultrasonic-microwave synchronization assisting ion liquid system preprocessing lignocellulose according to claim 8 is improved
The method of its enzymolysis efficiency, it is characterised in that add cellulose enzyme amount to be 5～1000U/g substrates in feed liquid.
10. ultrasonic-microwave synchronization assisting ion liquid system preprocessing lignocellulose according to claim 1 is improved
The method of its enzymolysis efficiency, it is characterised in that the cellulase is that commercially available extensive stock enzyme preparation or microorganism are direct
The enzyme liquid that fermentation is obtained.
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