CN107177644A - A kind of method of utilization ultrasonic assistant ionic liquid preprocessing biomass - Google Patents
A kind of method of utilization ultrasonic assistant ionic liquid preprocessing biomass Download PDFInfo
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- CN107177644A CN107177644A CN201710354461.2A CN201710354461A CN107177644A CN 107177644 A CN107177644 A CN 107177644A CN 201710354461 A CN201710354461 A CN 201710354461A CN 107177644 A CN107177644 A CN 107177644A
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- C12P19/00—Preparation of compounds containing saccharide radicals
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Abstract
The invention discloses a kind of method of utilization ultrasonic assistant ionic liquid preprocessing biomass, belong to biomass utilization technologies field.Biomass is pre-processed from the methylimidazole villaumite of 1 butyl of anion imidazole ion liquid 3 and the methylimidazole acetate of 1 butyl 3 first, apply the ultrasonic wave of five frequencies respectively simultaneously to aid in pretreatment, then plus distillation dissolving, through filtering, being dried to obtain renewable biomass, enzyme hydrolysis and sour water solution then are carried out to renewable biomass under certain condition.Yield, including enzyme hydrolysis yield and sour water solution yield are hydrolyzed the method can significantly improve the reduced sugar of biomass, Reducing sugar improves 27.5% ~ 38.1%.Regenerate and reuse with ionic liquid, advantage and the low energy consumption such as ultrasonic wave green physical field, low cost are simple to operate, the features such as environment-friendly.
Description
Technical field
The invention belongs to biomass utilization technologies field, and in particular to one kind utilizes ultrasonic assistant ionic liquid pretreatment
The method that biomass improves its percent hydrolysis.
Background technology
As economic develops rapidly, the rapid growth of population, the mankind increase the demand of the energy increasingly, non-renewable
Fossil resources are just depleted and face increasingly serious environmental problem, seriously constrain whole world sustainable development,
The renewable resource that exploitation cleaning substitutes petroleum resources is extremely urgent.Biomass is unique on the earth converted by solar energy
A kind of rich reserves the renewable resource for storing and transporting.Lignocellulosic is the chief component of biomass, its
Be widely present in agricultural crop straw, yield is big, be easy to get to and the advantage such as cheap become most Exploitative potential
One of biomass resource, produce reduced sugar using stalk in recent years and also important grind for one as countries in the world biomass economy
Study carefully direction.
Ionic liquid has solvability strong because of it, has good dissolubility energy to organic and inorganic compound；Liquid
State temperature scope is wide, and heat endurance is high；Easily reclaim, can be recycled；With designability, it can be constituted by adjusting anion
The features such as physics and chemical property to change itself, is as in recent years as the main pretreating agent of cellulose, but be due to
Ionic liquid costly, with certain toxicity, can be lost in dissolving cellulose and removal process, and right after pretreatment
Cellulose percent hydrolysis improve it is limited, so on this basis introduce ultrasonic assistant ionic liquid pretreatment biomass with increase from
Sub- liquid reduces the usage amount of ionic liquid as the solubility effect of pretreating agent, saves cost.
Ultrasonic wave is one kind of sound wave, because it can produce cavitation effect, fuel factor, mechanical effect etc. in communication process
Effect, also gradually applies to the degraded of macromolecular substances, in terms of such as polysaccharide degraded and proteolysis.At present in biomass degradation
There has been a certain degree of research in field.Cavitation and machine that the effect performance that ultrasonic wave is pre-processed to biomass is produced in the solution
Tool effect can be impacted and be sheared to biological surface, can effectively destroy hydrogen bond in cellulosic molecule, reduce crystallinity,
So that biomass becomes coarse and irregular, more contact sites are provided in enzyme hydrolysis and sour water solution, less hydrogen bond is sheared.
Therefore, introducing ultrasonic assistant ionic liquid makes biomass further hydrolyze, and improves resource utilization.
The content of the invention
It is an object of the invention to provide one kind its acid or enzyme are improved using ultrasonic assistant ionic liquid pretreatment biomass
The method of percent hydrolysis.
The method that utilization ultrasonic assistant ionic liquid pretreatment biomass proposed by the present invention improves its percent hydrolysis, first
From anion imidazole ion liquid 1- butyl -3- methylimidazole villaumites and 1- butyl -3- methylimidazole acetate to biomass
Pre-processed, while applying the ultrasonic wave of five frequencies respectively to aid in pre-processing, then add distillation dissolving, through filtering, do
It is dry to obtain renewable biomass, enzyme hydrolysis and sour water solution then are carried out to renewable biomass under certain condition, finally using efficient
Liquid chromatography and DNS methods determine the content of monose and reduced sugar in hydrolyzate, and compare before and after pretreatment and individually make
With the monose and content of reducing sugar of ionic liquid pretreatment, ultrasonic assistant ionic liquid pretreatment biomass is studied to it with this
The influence of percent hydrolysis.
To achieve the above object, the present invention is adopted the following technical scheme that：One kind utilizes the pretreatment of ultrasonic assistant ionic liquid
The method of biomass, is carried out as steps described below：
（1）Using ionic liquid as solvent, according to 1:10~1:20 solid-to-liquid ratio（g/mL）Biomass is mixed with ionic liquid,
The magnetic agitation at 70 ~ 80 DEG C, and applying the min of ultrasonic wave 20 ~ 30 of different frequency makes biomass be dissolved completely in ionic liquid
In；
（2）After question response terminates, distilled water is added in reaction solution according to the amount of 3 ~ 5 times of volumes of ionic liquid, ionic liquid
Body is dissolved into distilled water and separates out precipitation, filters out precipitation and with water washing is distilled 3 ~ 5 times after 80 DEG C of drying of baking oven, obtains
Renewable biomass；
（3）Ionic liquid is reclaimed using the method for vacuum distillation.After biomass Precipitation, obtained solution be water and
Ionic liquid mixing liquid, rotary evaporation removes moisture removal under the conditions of 60 DEG C, 100 rpm, until moisture not re-evaporation, and 80
12 ~ 24 h are dried under the conditions of DEG C to remove residual moisture.
（4）The pretreatment renewable biomass of certain mass is weighed, according to solid-to-liquid ratio 1:300~1:500 (g/ml) are added to
With in the cellulase solution of Acetic acid-sodium acetate cushion, as in shaking table, reacting 12 under 50 ~ 60 DEG C, 80 ~ 150 rpm ~
24 h, obtain enzyme hydrolyzate；
（5）The pretreatment renewable biomass of certain mass is weighed, according to solid-to-liquid ratio 1:20 (g/mL) and mass fraction for 2% it is dilute
60 ~ 90 min are reacted after sulfuric acid mixing at 160 ~ 180 DEG C, reaction solution is removed by filtration precipitation after terminating, will filtered by reaction
Liquid is settled to a certain amount of, the sour hydrating solution of acquisition；
（6）Hydrolysis determines sugared monose in enzyme hydrolyzate and acid hydrolysis liquid and content of reducing sugar and calculated respectively after terminating
Respective yield.
In the present invention, the ionic liquid used is anion imidazole ion liquid 1- butyl -3- methylimidazole villaumites
（[Bmim]Cl）With 1- butyl -3- methylimidazole acetate（[Bmim]OAc）.
In the present invention, biomass used includes agricultural waste material：Such as rice straw, wheat stalk, maize straw；Agriculture
Product processing discarded object：Such as bagasse, sugarcane skin, Cortex Bulbus Allii.
In the present invention, used ultrasonic device is energy gathering type single-frequency ultrasonic equipment.
In the present invention, supersonic frequency used includes 20,28,35,40 and 50kHz, five frequencies, and ultrasonic power is 100 W
Within some firm power.
In the present invention, ultrasonic probe is stretched into level control a certain constant depth in 1 ~ 3 cm when ultrasonic device is used
Degree.
In the present invention, firing equipment is thermostatic water-circulator bath pot, supports the use reaction vessel for double-jacket beaker.
In the present invention, precipitation is soaked in ethanol before cleaning precipitation and 15 ~ 30 min are stirred.
In the present invention, it is cloth funnel and rotary-vane vaccum pump to filter the instrument used in precipitation.
In the present invention, Acetic acid-sodium acetate buffer solution ph used is 4.8 during enzyme hydrolysis.
In the present invention, acid hydrolytic reaction container is the autoclave of each model, including Pressure solution bullet.
In the present invention, hydrolyzate contents of monosaccharides detection method utilizes high performance liquid chromatography, and content of reducing sugar utilizes DNS
Method.
The invention has the advantages that：
（1）Compared with traditional preprocessing solution, the ionic liquid used in the present invention has dissolubility strong, designability
By force, solvability is strong, and chemical stability is good, environment-friendly and with recuperability, meets sustainable green chemistry
It is required that.
（2）[Bmim] Cl and two kinds of ionic liquids of [Bmim] OAc in the present invention have recuperability, and the rate of recovery is 80%
More than, and reclaim after ionic liquid maintain original performance, can be used with secondary.
（3）Ultrasonic assistant ionic liquid pretreatment biomass is introduced in the present invention during pretreatment, because of its tool
There are the characteristics such as pack, orientation and reflection, transmission, can effectively open the crystal structure of lignocellulosic, and destroy wooden
Element, makes biomass more facile hydrolysis.
（4）The ultrasonic device used in the present invention is energy gathering type single-frequency ultrasonic wave, can control to pop one's head in and go deep into the depth of liquid level
The features such as degree, supersonic frequency, ultrasonic power, ultrasonic time and pulse retention time and intermittent time, it can efficiently control
The ultrasound condition of reaction.
（5）The firing equipment used in the present invention is thermostatic water-circulator bath pot, is heated compared to traditional water bath with thermostatic control, this
Equipment heated for controlling temperature is more accurate, can control, at ± 0.1 DEG C, to carry out using stable reaction.
（6）Reaction vessel used in the present invention is double-jacket beaker, coordinates thermostatic water-circulator bath pot, can be effective
It is stable that reaction environment is provided, and can be with observing response phenomenon.
（7）Monosaccharide quantitation is carried out to hydrolyzate using high performance liquid chromatography in the present invention, the chromatographic column used is the U.S.
The Aminex of Bio-Rad companies production@HPX-87H chromatographic columns.It can efficiently separate and quantify hydrolyzate monose.
Embodiment
Illustrate the present invention by embodiment, but do not invent and be not limited only to embodiment.
Embodiment 1：
（1）Take the g of [Bmim] Cl ionic liquids 20 into double-jacket beaker, 1 g bagasse is slowly added into ionic liquid
In, using thermostatic water-circulator bath pot 80 DEG C of temperature of control, energy gathering type ultrasonic probe is stretched into the cm of liquid level 2, the W of ultrasonic power 100,
The kHz of supersonic frequency 20, the burst length keeps 10 s, the s of intermittent time 2, on this condition ultrasonic 30 min.After pretreatment terminates
Plus 3 ~ distilled water of 5 times of original solutions produces precipitation, allows precipitation to be dissolved in after filtering in 20 ~ 30 mL ethanol, magnetic agitation 20
Min, is precipitated to remove survivor ion liquid with the distilled water flushing of 5 ~ 8 times of volumes of original solution after filtering, obtained precipitation is dried
Obtain regenerating bagasse after dry.
（2）The cushioning liquid that pH is 4.8 is made into using acetic acid, sodium acetate as cushion, then uses buffer cellulase
Liquid（1 g/L）, 50 mL cellulase solutions are taken into 150 mL conical flasks, 0.1 g regeneration bagasse are added, in 50 DEG C of shaking tables
24 h of middle reaction, filter, filtrate are settled into 100 mL with distilled water, cryopreservation is to be measured afterwards.
（3）Accurate to weigh drying to the regeneration bagasse of constant weight, the dilute sulfuric acid with 2% is according to 20:1 （mL/g）It is sufficiently mixed
After be transferred in reactor, 160 DEG C of 90 min of heating, reaction filters to take supernatant 100 mL is settled to distilled water after terminating,
Cryopreservation is to be measured.
（4）Content of reducing sugar in hydrolyzate is determined using DNS methods.Enzyme hydrolysis and each 0.5 mL of sour water solution prepare liquid are taken, plus
Enter and be cooled to room temperature after reacting 15 min, taking-up in 1.5 mL DNS agent and 1.5 mL distilled water, boiling water bath, be settled to 20 mL,
Under 54 nm absorbance is determined under spectrophotometric.It is respectively 77.85% He finally to determine enzyme hydrolysis and sour water solution Reducing sugar
70.40%
（5）Utilize contents of monosaccharides in Water By High Performance Liquid solution liquid.Utilize the Agilent 1260 for being equipped with Composition distribution
High performance liquid chromatograph, Aminex@HPX-87H chromatographic columns, 60 DEG C of column temperature, 50 DEG C of detector, 5 mM H2SO4Surveyed as mobile phase
Determine glucose, xylose and the arabinose in cellobiose, the glucose and xylose in enzyme hydrolyzate, acid hydrolysis liquid.Measure enzyme
Hydrolysis fiber disaccharides, glucose, xylose yield are respectively 3.82%, 40.32%, 12.53%；Sour hydrolyzation of glucose, xylose, I
Uncle's sugar 30.94%, 24.14%, 2.24%.
Embodiment 2：
（1）Take the g of [Bmim] Cl ionic liquids 10 into double-jacket beaker, 1 g bagasse is slowly added into ionic liquid
In, using thermostatic water-circulator bath pot control temperature 70 C, energy gathering type ultrasonic probe is stretched into the cm of liquid level 2, the W of ultrasonic power 100,
The kHz of supersonic frequency 20, the burst length keeps 10 s, the s of intermittent time 2, on this condition ultrasonic 20 min.After pretreatment terminates
Plus 3 ~ distilled water of 5 times of original solutions produces precipitation, allows precipitation to be dissolved in after filtering in 20 ~ 30 mL ethanol, magnetic agitation 20
Min, is precipitated to remove survivor ion liquid with the distilled water flushing of 5 ~ 8 times of volumes of original solution after filtering, obtained precipitation is dried
Obtain regenerating bagasse after dry.
（2）The cushioning liquid that pH is 4.8 is made into using acetic acid, sodium acetate as cushion, then uses buffer cellulase
Liquid（1 g/L）, 30 mL cellulase solutions are taken into 250mL conical flasks, 0.1 g regeneration bagasse are added, in 50 DEG C of shaking tables
12 h are reacted, are filtered afterwards, filtrate is settled to 100 mL with distilled water, cryopreservation is to be measured.
（3）Accurate to weigh drying to the regeneration bagasse of constant weight, the dilute sulfuric acid with 2% is according to 20:1 （mL/g）It is sufficiently mixed
After be transferred in reactor, 180 DEG C of 60 min of heating, reaction filters to take supernatant 100 mL is settled to distilled water after terminating,
Cryopreservation is to be measured.
（4）Content of reducing sugar in hydrolyzate is determined using DNS methods.Enzyme hydrolysis and each 0.5 mL of sour water solution prepare liquid are taken, plus
Enter and be cooled to room temperature after reacting 15 min, taking-up in 1.5 mL DNS agent and 1.5 mL distilled water, boiling water bath, be settled to 20 mL,
Under 54 nm absorbance is determined under spectrophotometric.It is respectively 63.22% He finally to determine enzyme hydrolysis and sour water solution Reducing sugar
61.34%
（5）Utilize contents of monosaccharides in Water By High Performance Liquid solution liquid.Utilize the Agilent 1260 for being equipped with Composition distribution
High performance liquid chromatograph, Aminex@HPX-87H chromatographic columns, 60 DEG C of column temperature, 50 DEG C of detector, 5 mM H2SO4Surveyed as mobile phase
Determine glucose, xylose and the arabinose in cellobiose, the glucose and xylose in enzyme hydrolyzate, acid hydrolysis liquid.Measure enzyme
Hydrolysis fiber disaccharides, glucose, xylose yield are respectively 2.16%, 33.77%, 8.63%；Sour hydrolyzation of glucose, xylose, Arab
Sugar 18.53%, 21.24%, 1.13%.
Embodiment 3：
（1）Take the g of [Bmim] OAc ionic liquids 20 into double-jacket beaker, 1 g bagasse is slowly added into ionic liquid
In, using thermostatic water-circulator bath pot 80 DEG C of temperature of control, energy gathering type ultrasonic probe is stretched into the cm of liquid level 2, the W of ultrasonic power 100,
The kHz of supersonic frequency 28, the burst length keeps 10 s, the s of intermittent time 2, on this condition ultrasonic 30 min.After pretreatment terminates
Plus 3 ~ distilled water of 5 times of original solutions produces precipitation, allows precipitation to be dissolved in after filtering in 20 ~ 30 mL ethanol, magnetic agitation 20
Min, is precipitated to remove survivor ion liquid after filtering with the distilled water flushing of original solution 5 ~ 8, after obtained precipitation is dried
To regeneration bagasse.
（2）The cushioning liquid that pH is 4.8 is made into using acetic acid, sodium acetate as cushion, then uses buffer cellulase
Liquid（1 g/L）, 50 mL cellulase solutions are taken into 150 mL conical flasks, 0.1 g regeneration bagasse are added, in 50 DEG C of shaking tables
24 h of middle reaction, filter, filtrate are settled into 100 mL with distilled water, cryopreservation is to be measured afterwards.
（3）Accurate to weigh drying to the regeneration bagasse of constant weight, the dilute sulfuric acid with 2% is according to 20:1（mL/g）It is sufficiently mixed
After be transferred in reactor, 160 DEG C of 90 min of heating, reaction filters to take supernatant 100 mL is settled to distilled water after terminating,
Cryopreservation is to be measured.
（4）Content of reducing sugar in hydrolyzate is determined using DNS methods.Enzyme hydrolysis and each 0.5 mL of sour water solution prepare liquid are taken, plus
Enter and be cooled to room temperature after reacting 15 min, taking-up in 1.5 mL DNS agent and 1.5 mL distilled water, boiling water bath, be settled to 20 mL,
Under 54 nm absorbance is determined under spectrophotometric.It is respectively 87.57% He finally to determine enzyme hydrolysis and sour water solution Reducing sugar
75.26%
（5）Utilize contents of monosaccharides in Water By High Performance Liquid solution liquid.Utilize the Agilent 1260 for being equipped with Composition distribution
High performance liquid chromatograph, Aminex@HPX-87H chromatographic columns, 60 DEG C of column temperature, 50 DEG C of detector, 5 mM H2SO4Surveyed as mobile phase
Determine glucose, xylose and the arabinose in cellobiose, the glucose and xylose in enzyme hydrolyzate, acid hydrolysis liquid.Measure enzyme
Hydrolysis fiber disaccharides, glucose, xylose yield are respectively 7.04%, 47.13%, 16.52%；Sour hydrolyzation of glucose, xylose, I
Uncle's sugar 46.32%, 20.23%, 2.71%.
Embodiment 4：
（1）Take the g of [Bmim] OAc ionic liquids 10 into double-jacket beaker, 1g bagasse is slowly added into ionic liquid
In, using thermostatic water-circulator bath pot control temperature 70 C, energy gathering type ultrasonic probe is stretched into the cm of liquid level 2, the W of ultrasonic power 100,
The kHz of supersonic frequency 28, the burst length keeps 10 s, the s of intermittent time 2, on this condition ultrasonic 20 min.After pretreatment terminates
Plus 3 ~ distilled water of 5 times of original solution volumes produces precipitation, allows precipitation to be dissolved in after filtering in 20 ~ 30 mL ethanol, magnetic agitation 20
Min, is precipitated to remove survivor ion liquid with the distilled water flushing of 5 ~ 8 times of volumes of original solution after filtering, obtained precipitation is dried
Obtain regenerating bagasse after dry.
（2）The cushioning liquid that pH is 4.8 is made into using acetic acid, sodium acetate as cushion, then uses buffer cellulase
Liquid（1 g/L）, 30 mL cellulase solutions are taken into 150 ml conical flasks, 0.1 g regeneration bagasse are added, in 50 DEG C of shaking tables
12 h of middle reaction, filter, filtrate are settled into 100 mL with distilled water, cryopreservation is to be measured afterwards.
（3）Accurate to weigh drying to the regeneration bagasse of constant weight, the dilute sulfuric acid with 2% is according to 20:1 （mL/g）It is sufficiently mixed
After be transferred in reactor, 180 DEG C of 60 min of heating, reaction filters to take supernatant 100 mL is settled to distilled water after terminating,
Cryopreservation is to be measured.
（4）Content of reducing sugar in hydrolyzate is determined using DNS methods.Enzyme hydrolysis and each 0.5 mL of sour water solution prepare liquid are taken, plus
Enter and be cooled to room temperature after reacting 15min, taking-up in 1.5 mL DNS agent and 1.5 mL distilled water, boiling water bath, be settled to 20 mL,
Under 54 nm absorbance is determined under spectrophotometric.It is respectively 71.46% He finally to determine enzyme hydrolysis and sour water solution Reducing sugar
62.23%
（5）Utilize contents of monosaccharides in Water By High Performance Liquid solution liquid.Utilize the Agilent 1260 for being equipped with Composition distribution
High performance liquid chromatograph, Aminex@HPX-87H chromatographic columns, 60 DEG C of column temperature, 50 DEG C of detector, 5 mM H2SO4Surveyed as mobile phase
Determine glucose, xylose and the arabinose in cellobiose, the glucose and xylose in enzyme hydrolyzate, acid hydrolysis liquid.Measure enzyme
Hydrolysis fiber disaccharides, glucose, xylose yield are respectively 4.35%, 33.46%, 12.34%；Sour hydrolyzation of glucose, xylose, I
Uncle's sugar 24.14%, 12.35%, 1.02%.
Embodiment 5：
（1）Take the g of [Bmim] Cl ionic liquids 20 into double-jacket beaker, 1 g wheat stalks are slowly added into ionic liquid
In, using thermostatic water-circulator bath pot 80 DEG C of temperature of control, energy gathering type ultrasonic probe is stretched into the cm of liquid level 2, the W of ultrasonic power 100,
The kHz of supersonic frequency 20, the burst length keeps 10 s, the s of intermittent time 2, on this condition ultrasonic 30 min.After pretreatment terminates
Plus 3 ~ distilled water of 5 times of original solutions produces precipitation, allows precipitation to be dissolved in after filtering in 20 ~ 30 mL ethanol, magnetic agitation 20
Min, is precipitated to remove survivor ion liquid with the distilled water flushing of 5 ~ 8 times of volumes of original solution after filtering, obtained precipitation is dried
Obtain regenerating bagasse after dry.
（2）The cushioning liquid that pH is 4.8 is made into using acetic acid, sodium acetate as cushion, then uses buffer cellulase
Liquid（1g/L）, 50 mL cellulase solutions are taken into 150 mL conical flasks, 0.1 g regeneration wheat stalks are added, in 50 DEG C of shaking tables
24 h of middle reaction, filter, filtrate are settled into 100 mL with distilled water, cryopreservation is to be measured afterwards.
（3）Accurate to weigh drying to the regeneration wheat stalk of constant weight, the dilute sulfuric acid with 2% is according to 20:1（mL/g）It is fully mixed
It is transferred to after conjunction in reactor, 160 DEG C of 90 min of heating, reaction filters to take supernatant after terminating and is settled to 100 with distilled water
ML, cryopreservation is to be measured.
（4）Content of reducing sugar in hydrolyzate is determined using DNS methods.Enzyme hydrolysis and each 0.5 mL of sour water solution prepare liquid are taken, plus
Enter and be cooled to room temperature after reacting 15 min, taking-up in 1.5 mL DNS agent and 1.5 mL distilled water, boiling water bath, be settled to 20 mL,
Under 54 nm absorbance is determined under spectrophotometric.It is respectively 91.89% He finally to determine enzyme hydrolysis and sour water solution Reducing sugar
73.27%
（5）Utilize contents of monosaccharides in Water By High Performance Liquid solution liquid.Utilize the Agilent 1260 for being equipped with Composition distribution
High performance liquid chromatograph, Aminex@HPX-87H chromatographic columns, 60 DEG C of column temperature, 50 DEG C of detector, 5 mM H2SO4Surveyed as mobile phase
Determine glucose, xylose and the arabinose in cellobiose, the glucose and xylose in enzyme hydrolyzate, acid hydrolysis liquid.Measure enzyme
Hydrolysis fiber disaccharides, glucose, xylose yield are respectively 9.25%, 53.17%, 18.13%；Sour hydrolyzation of glucose, xylose, I
Uncle's sugar 48.07%, 19.26%, 1.85%.
Embodiment 6：
（1）Take the g of [Bmim] Cl ionic liquids 10 into double-jacket beaker, 1 g wheat stalks are slowly added into ionic liquid
In, using thermostatic water-circulator bath pot control temperature 70 C, energy gathering type ultrasonic probe is stretched into the cm of liquid level 2, the W of ultrasonic power 100,
The kHz of supersonic frequency 20, the burst length keeps 10 s, the s of intermittent time 2, on this condition ultrasonic 10 min.After pretreatment terminates
Plus 3 ~ distilled water of 5 times of original solution volumes produces precipitation, allows precipitation to be dissolved in after filtering in 20 ~ 30 mL ethanol, magnetic agitation 20
Min, is precipitated to remove survivor ion liquid with the distilled water flushing of 5 ~ 8 times of volumes of original solution after filtering, obtained precipitation is dried
Obtain regenerating bagasse after dry.
（2）The cushioning liquid that pH is 4.8 is made into using acetic acid, sodium acetate as cushion, then uses buffer cellulase
Liquid（1 g/L）, 20 mL cellulase solutions are taken into 150 mL conical flasks, are added 0.1 g regeneration wheat stalks, are shaken at 50 DEG C
12 h are reacted in bed, filters afterwards, filtrate is settled to 100 mL with distilled water, cryopreservation is to be measured.
（3）Accurate to weigh drying to the regeneration wheat stalk of constant weight, the dilute sulfuric acid with 2% is according to 20:1（mL/g）It is fully mixed
It is transferred to after conjunction in reactor, 180 DEG C of 60 min of heating, reaction filters to take supernatant after terminating and is settled to 100 with distilled water
ML, cryopreservation is to be measured.
（4）Content of reducing sugar in hydrolyzate is determined using DNS methods.Enzyme hydrolysis and each 0.5 mL of sour water solution prepare liquid are taken, plus
Enter and be cooled to room temperature after reacting 15 min, taking-up in 1.5 mL DNS agent and 1.5 mL distilled water, boiling water bath, be settled to 20 mL,
Under 54 nm absorbance is determined under spectrophotometric.It is respectively 82.13% He finally to determine enzyme hydrolysis and sour water solution Reducing sugar
65.32%
（5）Utilize contents of monosaccharides in Water By High Performance Liquid solution liquid.Utilize the Agilent 1260 for being equipped with Composition distribution
High performance liquid chromatograph, Aminex@HPX-87H chromatographic columns, 60 DEG C of column temperature, 50 DEG C of detector, 5 mM H2SO4Surveyed as mobile phase
Determine glucose, xylose and the arabinose in cellobiose, the glucose and xylose in enzyme hydrolyzate, acid hydrolysis liquid.Measure enzyme
Hydrolysis fiber disaccharides, glucose, xylose yield are respectively 5.53%, 38.57%, 12.84%；Sour hydrolyzation of glucose, xylose, I
Uncle's sugar 34.23%, 12.58%, 1.29%.
Embodiment 7：
（1）Take the g of [Bmim] OAc ionic liquids 20 into double-jacket beaker, 1 g wheat stalks are slowly added into ionic liquid
In body, using thermostatic water-circulator bath pot 80 DEG C of temperature of control, energy gathering type ultrasonic probe is stretched into the cm of liquid level 2, ultrasonic power 100
W, the kHz of supersonic frequency 28, the burst length keep 10 s, the s of intermittent time 2, on this condition ultrasonic 30 min.Pretreatment terminates
Plus 3 afterwards ~ distilled water of 5 times of original solutions produces precipitation, allows precipitation to be dissolved in after filtering in 20 ~ 30 mL ethanol, magnetic agitation 20
Min, is precipitated to remove survivor ion liquid with the distilled water flushing of 5 ~ 8 times of volumes of original solution after filtering, obtained precipitation is dried
Obtain regenerating wheat stalk after dry.
（2）The cushioning liquid that pH is 4.8 is made into using acetic acid, sodium acetate as cushion, then uses buffer cellulase
Liquid（1 g/L）, 50ml cellulase solutions are taken into 150 mL conical flasks, 0.1 g regeneration wheat stalks are added, in 50 DEG C of shaking tables
24 h of middle reaction, filter, filtrate are settled into 100 mL with distilled water, cryopreservation is to be measured afterwards.
（3）Accurate to weigh drying to the regeneration wheat stalk of constant weight, the dilute sulfuric acid with 2% is according to 20:1（mL/g）It is fully mixed
It is transferred to after conjunction in reactor, 160 DEG C of 90 min of heating, reaction filters to take supernatant after terminating and is settled to 100 with distilled water
ML, cryopreservation is to be measured.
（4）Content of reducing sugar in hydrolyzate is determined using DNS methods.Enzyme hydrolysis and each 0.5 mL of sour water solution prepare liquid are taken, plus
Enter and be cooled to room temperature after reacting 15 min, taking-up in 1.5 mL DNS agent and 1.5 mL distilled water, boiling water bath, be settled to 20 mL,
Under 54 nm absorbance is determined under spectrophotometric.It is respectively 93.68% He finally to determine enzyme hydrolysis and sour water solution Reducing sugar
76.70%
（5）Utilize contents of monosaccharides in Water By High Performance Liquid solution liquid.Utilize the Agilent 1260 for being equipped with Composition distribution
High performance liquid chromatograph, Aminex@HPX-87H chromatographic columns, 60 DEG C of column temperature, 50 DEG C of detector, 5 mM H2SO4Surveyed as mobile phase
Determine glucose, xylose and the arabinose in cellobiose, the glucose and xylose in enzyme hydrolyzate, acid hydrolysis liquid.Measure enzyme
Hydrolysis fiber disaccharides, glucose, xylose yield are respectively 8.61%, 54.25%, 12.90%；Sour hydrolyzation of glucose, xylose, I
Uncle's sugar 43.73%, 19.10%, 2.45%.
Embodiment 8：
（1）Take the g of [Bmim] OAc ionic liquids 10 into double-jacket beaker, 1 g wheat stalks are slowly added into ionic liquid
In body, using thermostatic water-circulator bath pot control temperature 70 C, energy gathering type ultrasonic probe is stretched into the cm of liquid level 2, ultrasonic power 100
W, the kHz of supersonic frequency 28, the burst length keep 10 s, the s of intermittent time 2, on this condition ultrasonic 20 min.Pretreatment terminates
Plus 3 afterwards ~ distilled water of 5 times of original solution volumes produces precipitation, allows precipitation to be dissolved in after filtering in 20 ~ 30 mL ethanol, magnetic agitation
20 min, are precipitated to remove survivor ion liquid after filtering with the distilled water flushing of 5 ~ 8 times of volumes of original solution, by obtained precipitation
Obtain regenerating wheat stalk after drying.
（2）The cushioning liquid that pH is 4.8 is made into using acetic acid, sodium acetate as cushion, then uses buffer cellulase
Liquid（1 g/L）, 30 mL cellulase solutions are taken into 150 mL conical flasks, are added 0.1 g regeneration wheat stalks, are shaken at 50 DEG C
12 h are reacted in bed, filters afterwards, filtrate is settled to 100 mL with distilled water, cryopreservation is to be measured.
（3）Accurate to weigh drying to the regeneration wheat stalk of constant weight, the dilute sulfuric acid with 2% is according to 20:1（mL/g）It is fully mixed
It is transferred to after conjunction in reactor, 180 DEG C of 60 min of heating, reaction filters to take supernatant after terminating and is settled to 100 with distilled water
ML, cryopreservation is to be measured.
（4）Content of reducing sugar in hydrolyzate is determined using DNS methods.Enzyme hydrolysis and each 0.5 mL of sour water solution prepare liquid are taken, plus
Enter and be cooled to room temperature after reacting 15 min, taking-up in 1.5 mL DNS agent and 1.5 mL distilled water, boiling water bath, be settled to 20 mL,
Under 54 nm absorbance is determined under spectrophotometric.It is respectively 82.75% He finally to determine enzyme hydrolysis and sour water solution Reducing sugar
58.92%
（5）Utilize contents of monosaccharides in Water By High Performance Liquid solution liquid.Utilize the Agilent 1260 for being equipped with Composition distribution
High performance liquid chromatograph, Aminex@HPX-87H chromatographic columns, 60 DEG C of column temperature, 50 DEG C of detector, 5 mM H2SO4Surveyed as mobile phase
Determine glucose, xylose and the arabinose in cellobiose, the glucose and xylose in enzyme hydrolyzate, acid hydrolysis liquid.Measure enzyme
Hydrolysis fiber disaccharides, glucose, xylose yield are respectively 5.63%, 37.62%, 8.38%；Sour hydrolyzation of glucose, xylose, Arab
Sugar 12.26%, 11.35%, 0.88%.
Reference examples 1：
Not pretreated bagasse enzyme hydrolysis and sour water solution
（1）The cushioning liquid that PH is 4.8 is made into using acetic acid, sodium acetate as cushion, then uses buffer cellulase solution
（1g/L）, 50 mL cellulase solutions are taken into 150 mL conical flasks, the not pretreated bagasse of 0.1 g are added, at 50 DEG C
24 h are reacted in shaking table, are filtered afterwards, filtrate is settled to 100 mL with distilled water, cryopreservation is to be measured.
（2）Accurate to weigh drying to the not pretreated bagasse of constant weight, the dilute sulfuric acid with 2% is according to 20:1（mL/g）
It is transferred to after being sufficiently mixed in reactor, 160 DEG C of 90 min of heating, reaction is filtered to take supernatant after terminating and is settled to distilled water
100 mL, cryopreservation is to be measured.
（3）Content of reducing sugar in hydrolyzate is determined using DNS methods.Enzyme hydrolysis and each 0.5 mL of sour water solution prepare liquid are taken, plus
Enter and be cooled to room temperature after reacting 15 min, taking-up in 1.5 mL DNS agent and 1.5 mL distilled water, boiling water bath, be settled to 20 mL,
Under 54 nm absorbance is determined under spectrophotometric.It is respectively 41.88% He finally to determine enzyme hydrolysis and sour water solution Reducing sugar
44.86%
（4）Utilize contents of monosaccharides in Water By High Performance Liquid solution liquid.Utilize the Agilent 1260 for being equipped with Composition distribution
High performance liquid chromatograph, Aminex@HPX-87H chromatographic columns, 60 DEG C of column temperature, 50 DEG C of detector, 5 mM H2SO4Surveyed as mobile phase
Determine glucose, xylose and the arabinose in cellobiose, the glucose and xylose in enzyme hydrolyzate, acid hydrolysis liquid.Measure enzyme
Hydrolysis fiber disaccharides, glucose, xylose yield are respectively 0.81%, 26.09%, 3.12%；Sour hydrolyzation of glucose, xylose, Arab
Sugar 8.82%, 4.25%, 0.55%.
Reference examples 2：
Only by [Bmim] Cl bagasse enzyme hydrolysis pre-processed and sour water solution
（1）Take the g of [Bmim] Cl ionic liquids 20 into double-jacket beaker, 1 g bagasse is slowly added into ionic liquid
In, using thermostatic water-circulator bath pot 80 DEG C of temperature of control, 30 min are dissolved on this condition.Pretreatment Jia 3 ~ 5 times of originals after terminating
The distilled water of liquor capacity produces precipitation, allows precipitation to be dissolved in after filtering in 20 ~ 30 mL ethanol, the min of magnetic agitation 20, filtering
Precipitated afterwards with the distilled water flushing of original solution 5 ~ 8 to remove survivor ion liquid, regeneration is obtained after obtained precipitation is dried sweet
Bagasse.
（2）The cushioning liquid that pH is 4.8 is made into using acetic acid, sodium acetate as cushion, then uses buffer cellulase
Liquid（1 g/L）, 50 mL cellulase solutions are taken into 150 mL conical flasks, 0.1 g regeneration bagasse are added, in 50 DEG C of shaking tables
24 h of middle reaction, filter, filtrate are settled into 100 mL with distilled water, cryopreservation is to be measured afterwards.
（3）Accurate to weigh drying to the regeneration bagasse of constant weight, the dilute sulfuric acid with 2% is according to 20:1（mL/g）It is sufficiently mixed
After be transferred in reactor, 160 DEG C of 90 min of heating, reaction filters to take supernatant 100 mL is settled to distilled water after terminating,
Cryopreservation is to be measured.
（4）Content of reducing sugar in hydrolyzate is determined using DNS methods.Enzyme hydrolysis and each 0.5 mL of sour water solution prepare liquid are taken, plus
Enter and be cooled to room temperature after reacting 15 min, taking-up in 1.5 mL DNS agent and 1.5 mL distilled water, boiling water bath, be settled to 20 mL,
Under 54 nm absorbance is determined under spectrophotometric.It is respectively 53.39% He finally to determine enzyme hydrolysis and sour water solution Reducing sugar
56.19%
（5）Utilize contents of monosaccharides in Water By High Performance Liquid solution liquid.Utilize the Agilent 1260 for being equipped with Composition distribution
High performance liquid chromatograph, Aminex@HPX-87H chromatographic columns, 60 DEG C of column temperature, 50 DEG C of detector, 5 mM H2SO4Surveyed as mobile phase
Determine glucose, xylose and the arabinose in cellobiose, the glucose and xylose in enzyme hydrolyzate, acid hydrolysis liquid.Measure enzyme
Hydrolysis fiber disaccharides, glucose, xylose yield are respectively 2.69%, 29.27%, 6.18%；Sour hydrolyzation of glucose, xylose, Arab
Sugar 21.78%, 8.70%, 1.01%.
Reference examples 3：
Only by [Bmim] OAc bagasse enzyme hydrolysis pre-processed and sour water solution
1）Take the g of [Bmim] OAc ionic liquids 20 into double-jacket beaker, 1 g bagasse is slowly added into ionic liquid
In, using thermostatic water-circulator bath pot 80 DEG C of temperature of control, 30 min are dissolved on this condition.Pretreatment Jia 3 ~ 5 times of originals after terminating
The distilled water of liquor capacity produces precipitation, allows precipitation to be dissolved in after filtering in 20 ~ 30 mL ethanol, the min of magnetic agitation 20, filtering
Precipitated afterwards with the distilled water flushing of original solution 5 ~ 8 to remove survivor ion liquid, regeneration is obtained after obtained precipitation is dried sweet
Bagasse.
（2）The cushioning liquid that pH is 4.8 is made into using acetic acid, sodium acetate as cushion, then uses buffer cellulase
Liquid（1 g/L）, 50 mL cellulase solutions are taken into 150 mL conical flasks, 0.1 g regeneration bagasse are added, in 50 DEG C of shaking tables
24 h of middle reaction, filter, filtrate are settled into 100 mL with distilled water, cryopreservation is to be measured afterwards.
（3）Accurate to weigh drying to the regeneration bagasse of constant weight, the dilute sulfuric acid with 2% is according to 20:1（mL/g）It is sufficiently mixed
After be transferred in reactor, 160 DEG C of 90 min of heating, reaction filters to take supernatant 100 mL is settled to distilled water after terminating,
Cryopreservation is to be measured.
（4）Content of reducing sugar in hydrolyzate is determined using DNS methods.Enzyme hydrolysis and each 0.5 mL of sour water solution prepare liquid are taken, plus
Enter and be cooled to room temperature after reacting 15 min, taking-up in 1.5 mL DNS agent and 1.5 mL distilled water, boiling water bath, be settled to 20 mL,
Under 54 nm absorbance is determined under spectrophotometric.It is respectively 52.18% He finally to determine enzyme hydrolysis and sour water solution Reducing sugar
50.26%
（5）Utilize contents of monosaccharides in Water By High Performance Liquid solution liquid.Utilize the Agilent 1260 for being equipped with Composition distribution
High performance liquid chromatograph, Aminex@HPX-87H chromatographic columns, 60 DEG C of column temperature, 50 DEG C of detector, 5 mM H2SO4Surveyed as mobile phase
Determine glucose, xylose and the arabinose in cellobiose, the glucose and xylose in enzyme hydrolyzate, acid hydrolysis liquid.Measure enzyme
Hydrolysis fiber disaccharides, glucose, xylose yield are respectively 3.20%, 32.73%, 6.77%；Sour hydrolyzation of glucose, xylose, Arab
Sugar 21.35%, 8.77%, 0.92%.
Reference examples 4：
Not pretreated wheat stalk enzyme hydrolysis and sour water solution
（1）The cushioning liquid that pH is 4.8 is made into using acetic acid, sodium acetate as cushion, then uses buffer cellulase solution（1
g/L）, 50 mL cellulase solutions are taken into 150 mL conical flasks, the not pretreated wheat stalks of 0.1 g are added, at 50 DEG C
24 h are reacted in shaking table, are filtered afterwards, filtrate is settled to 100 mL with distilled water, cryopreservation is to be measured.
（2）Accurate to weigh drying to the not pretreated wheat stalk of constant weight, the dilute sulfuric acid with 2% is according to 20:1（mL/
g）It is transferred to after being sufficiently mixed in reactor, 160 DEG C of 90 min of heating, reaction filters to take supernatant distilled water constant volume after terminating
To 100 mL, cryopreservation is to be measured.
（3）Content of reducing sugar in hydrolyzate is determined using DNS methods.Enzyme hydrolysis and each 0.5 mL of sour water solution prepare liquid are taken, plus
Enter and be cooled to room temperature after reacting 15 min, taking-up in 1.5 mL DNS agent and 1.5 mL distilled water, boiling water bath, be settled to 20 mL,
Under 54 nm absorbance is determined under spectrophotometric.Finally determine enzyme hydrolysis and sour water solution Reducing sugar is respectively:35.77% He
39.29%
（4）Utilize contents of monosaccharides in Water By High Performance Liquid solution liquid.Utilize the Agilent 1260 for being equipped with Composition distribution
High performance liquid chromatograph, Aminex@HPX-87H chromatographic columns, 60 DEG C of column temperature, 50 DEG C of detector, 5 mM H2SO4Surveyed as mobile phase
Determine glucose, xylose and the arabinose in cellobiose, the glucose and xylose in enzyme hydrolyzate, acid hydrolysis liquid.Measure enzyme
Hydrolysis fiber disaccharides, glucose, xylose yield are respectively 1.49%, 23.63%, 2.57%；Sour hydrolyzation of glucose, xylose, Arab
Sugar 8.19%, 3.49%, 0.31%.
Reference examples 5：
Only by [Bmim] Cl wheat stalk enzyme hydrolysis pre-processed and sour water solution
（1）Take the g of [Bmim] Cl ionic liquids 20 into double-jacket beaker, 1 g wheat stalks are slowly added into ionic liquid
In, using thermostatic water-circulator bath pot 80 DEG C of temperature of control, 30 min are dissolved on this condition.Pretreatment Jia 3 ~ 5 times of originals after terminating
The distilled water of liquor capacity produces precipitation, allows precipitation to be dissolved in after filtering in 20 ~ 30 mL ethanol, the min of magnetic agitation 20, filtering
Precipitated afterwards with the distilled water flushing of original solution 5 ~ 8 to remove survivor ion liquid, regeneration is obtained after obtained precipitation is dried small
Wheat Straw.
（2）The cushioning liquid that pH is 4.8 is made into using acetic acid, sodium acetate as cushion, then uses buffer cellulase
Liquid（1 g/L）, 50 mL cellulase solutions are taken into 150 mL conical flasks, are added 0.1 g regeneration wheat stalks, are shaken at 50 DEG C
24 h are reacted in bed, filters afterwards, filtrate is settled to 100 mL with distilled water, cryopreservation is to be measured.
（3）Accurate to weigh drying to the regeneration wheat stalk of constant weight, the dilute sulfuric acid with 2% is according to 20:1（mL/g）It is fully mixed
It is transferred to after conjunction in reactor, 160 DEG C of 90 min of heating, reaction filters to take supernatant after terminating and is settled to 100 with distilled water
ML, cryopreservation is to be measured.
（4）Content of reducing sugar in hydrolyzate is determined using DNS methods.Enzyme hydrolysis and each 0.5 mL of sour water solution prepare liquid are taken, plus
Enter and be cooled to room temperature after reacting 15 min, taking-up in 1.5 mL DNS agent and 1.5 mL distilled water, boiling water bath, be settled to 20 mL,
Under 54 nm absorbance is determined under spectrophotometric.It is respectively 41.88% He finally to determine enzyme hydrolysis and sour water solution Reducing sugar
43.73%
（5）Utilize contents of monosaccharides in Water By High Performance Liquid solution liquid.Utilize the Agilent 1260 for being equipped with Composition distribution
High performance liquid chromatograph, Aminex@HPX-87H chromatographic columns, 60 DEG C of column temperature, 50 DEG C of detector, 5 mM H2SO4Surveyed as mobile phase
Determine glucose, xylose and the arabinose in cellobiose, the glucose and xylose in enzyme hydrolyzate, acid hydrolysis liquid.Measure enzyme
Hydrolysis fiber disaccharides, glucose, xylose yield are respectively 2.25%, 28.75%, 8.57%；Sour hydrolyzation of glucose, xylose, Arab
Sugar 17.41%, 9.58%, 1.14%.
Reference examples 6：
Only by [Bmim] OAc wheat stalk enzyme hydrolysis pre-processed and sour water solution
（1）Take the g of [Bmim] Cl ionic liquids 20 into double-jacket beaker, 1 g wheat stalks are slowly added into ionic liquid
In, using thermostatic water-circulator bath pot 80 DEG C of temperature of control, 30 min are dissolved on this condition.Pretreatment Jia 3 ~ 5 times of originals after terminating
The distilled water of liquor capacity produces precipitation, allows precipitation to be dissolved in after filtering in 20 ~ 30 mL ethanol, the min of magnetic agitation 20, filtering
Precipitated afterwards with the distilled water flushing of original solution 5 ~ 8 to remove survivor ion liquid, regeneration is obtained after obtained precipitation is dried small
Wheat Straw.
（2）The cushioning liquid that pH is 4.8 is made into using acetic acid, sodium acetate as cushion, then uses buffer cellulase
Liquid（1 g/L）, 50 mL cellulase solutions are taken into 150 mL conical flasks, are added 0.1 g regeneration wheat stalks, are shaken at 50 DEG C
24 h are reacted in bed, filters afterwards, filtrate is settled to 100 mL with distilled water, cryopreservation is to be measured.
（3）Accurate to weigh drying to the regeneration wheat stalk of constant weight, the dilute sulfuric acid with 2% is according to 20:1（mL/g）It is fully mixed
It is transferred to after conjunction in reactor, 160 DEG C of 90 min of heating, reaction filters to take supernatant after terminating and is settled to 100 with distilled water
ML, cryopreservation is to be measured.
（4）Content of reducing sugar in hydrolyzate is determined using DNS methods.Enzyme hydrolysis and each 0.5 mL of sour water solution prepare liquid are taken, plus
Enter and be cooled to room temperature after reacting 15 min, taking-up in 1.5 mL DNS agent and 1.5 mL distilled water, boiling water bath, be settled to 20 mL,
Under 54 nm absorbance is determined under spectrophotometric.It is respectively 49.44% He finally to determine enzyme hydrolysis and sour water solution Reducing sugar
49.00%
（5）Utilize contents of monosaccharides in Water By High Performance Liquid solution liquid.Utilize the Agilent 1260 for being equipped with Composition distribution
High performance liquid chromatograph, Aminex@HPX-87H chromatographic columns, 60 DEG C of column temperature, 50 DEG C of detector, 5 mM H2SO4Surveyed as mobile phase
Determine glucose, xylose and the arabinose in cellobiose, the glucose and xylose in enzyme hydrolyzate, acid hydrolysis liquid.Measure enzyme
Hydrolysis fiber disaccharides, glucose, xylose yield are respectively 2.23%, 29.89%, 4.57%；Sour hydrolyzation of glucose, xylose, Arab
Sugar 11.51%, 7.49%, 0.89%.
Embodiment is compared with reference examples, and reduced sugar total amount improves 27.5% ~ 38.1%.It is an advantage of the present invention that first,
[Bmim] Cl and two kinds of ionic liquids of [Bmim] OAc have recuperability, and the rate of recovery is more than 80%, and the ionic liquid after recovery
Body maintains original performance, can be used with secondary.Secondly, it is applied with ultrasonic assistant in biomass preprocessing process, and this
Individual process does not add any chemical substance, will not produce new chemical product during subsequent reactions.Last ultrasonic wave production
Raw cavitation effect and mechanical effect causes biomass not plan a successor, and surface is more coarse, and space increase, impalpable structure increases
Plus, this make it that enzyme hydrolysis and acid hydrolytic reaction are more abundant, so the present invention's is a kind of method of green high-efficient.
Claims (10)
1. a kind of method of utilization ultrasonic assistant ionic liquid preprocessing biomass, it is characterised in that carry out as steps described below：
（1）Using ionic liquid as solvent, according to 1:10~1:20 solid-to-liquid ratio（g/mL）Biomass is mixed with ionic liquid,
The magnetic agitation at 70 ~ 80 DEG C, and applying the min of ultrasonic wave 20 ~ 30 of different frequency makes biomass be dissolved completely in ionic liquid
In；
（2）After question response terminates, distilled water is added in reaction solution according to the amount of 3 ~ 5 times of volumes of ionic liquid, ionic liquid
Body is dissolved into distilled water and separates out precipitation, filters out precipitation and with water washing is distilled 3 ~ 5 times after 80 DEG C of drying of baking oven, obtains
Pre-process renewable biomass；
（3）Ionic liquid is reclaimed using the method for vacuum distillation；
After biomass Precipitation, obtained solution is water and ionic liquid mixing liquid, in 60 DEG C, 100 rpm condition backspins
Turn evaporative removal moisture, until moisture not re-evaporation, and dry 12 ~ 24 h to remove residual moisture under the conditions of 80 DEG C；
（4）The pretreatment renewable biomass of certain mass is weighed, according to solid-to-liquid ratio 1:300~1:500 (g/ml) are added to vinegar
In the cellulase solution of acid-sodium acetate buffer thing, as in shaking table, 12 ~ 24 h are reacted under 50 ~ 60 DEG C, 80 ~ 150 rpm,
Obtain enzyme hydrolyzate；
（5）The pretreatment renewable biomass of certain mass is weighed, according to solid-to-liquid ratio 1:20 (g/mL) and mass fraction for 2% it is dilute
60 ~ 90 min are reacted after sulfuric acid mixing at 160 ~ 180 DEG C, reaction solution is removed by filtration precipitation after terminating, will filtered by reaction
Liquid is settled to a certain amount of, the sour hydrating solution of acquisition；
（6）Hydrolysis determines sugared monose in enzyme hydrolyzate and acid hydrolysis liquid and content of reducing sugar and calculated respectively after terminating
Respective yield.
2. a kind of method of utilization ultrasonic assistant ionic liquid preprocessing biomass according to claim 1, its feature exists
In the ionic liquid used be anion imidazole ion liquid 1- butyl -3- methylimidazole villaumites（[Bmim]Cl）Or 1- butyl-
3- methylimidazole acetate（[Bmim]OAc）.
3. a kind of method of utilization ultrasonic assistant ionic liquid preprocessing biomass according to claim 1, its feature exists
Include agricultural waste material in biomass used：Such as rice straw, wheat stalk, maize straw；Processing of farm products discarded object：
Such as bagasse, sugarcane skin, Cortex Bulbus Allii.
4. a kind of method of utilization ultrasonic assistant ionic liquid preprocessing biomass according to claim 1, its feature exists
In used ultrasonic device be energy gathering type single-frequency ultrasonic equipment.
5. a kind of method of utilization ultrasonic assistant ionic liquid preprocessing biomass according to claim 4, its feature exists
Include 20,28,35,40 and 50 kHz, five frequencies in supersonic frequency used, ultrasonic power is constant for some within 100 W
Power.
6. a kind of method of utilization ultrasonic assistant ionic liquid preprocessing biomass according to claim 4, its feature exists
Ultrasonic probe is stretched into level control a certain constant depth in 1 ~ 3cm when ultrasonic device is used.
7. a kind of method of utilization ultrasonic assistant ionic liquid preprocessing biomass according to claim 1, its feature exists
It is thermostatic water-circulator bath pot in firing equipment, supports the use reaction vessel for double-jacket beaker.
8. a kind of method of utilization ultrasonic assistant ionic liquid preprocessing biomass according to claim 1, its feature exists
Precipitation is soaked and stirs 15 ~ 30 min in ethanol before cleaning is precipitated.
9. a kind of method of utilization ultrasonic assistant ionic liquid preprocessing biomass according to claim 1, its feature exists
Instrument used in being precipitated in filtering is cloth funnel and rotary-vane vaccum pump.
10. a kind of method of utilization ultrasonic assistant ionic liquid preprocessing biomass according to claim 1, its feature exists
Acetic acid-sodium acetate pH of buffer used is 4.8 when enzyme hydrolysis；Acid hydrolytic reaction container is the autoclave of each model,
Including Pressure solution bullet.
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| CN111154817A (en) * | 2020-02-19 | 2020-05-15 | 中国科学院过程工程研究所 | Method for efficiently separating lignocellulose and carrying out enzymolysis by using ionic liquid-high-boiling-point alcohol composite system |
| CN114431331A (en) * | 2021-12-31 | 2022-05-06 | 无锡金农生物科技有限公司 | Preparation method of high-water-solubility rice protein |
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| CN105255966A (en) * | 2015-10-27 | 2016-01-20 | 江苏大学 | Method for raising hydrolysis yield of microcrystalline cellulose through ionic liquid pretreatment |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111154817A (en) * | 2020-02-19 | 2020-05-15 | 中国科学院过程工程研究所 | Method for efficiently separating lignocellulose and carrying out enzymolysis by using ionic liquid-high-boiling-point alcohol composite system |
| CN111154817B (en) * | 2020-02-19 | 2022-04-08 | 中国科学院过程工程研究所 | Method for efficiently separating lignocellulose and carrying out enzymolysis by using ionic liquid-high-boiling-point alcohol composite system |
| CN114431331A (en) * | 2021-12-31 | 2022-05-06 | 无锡金农生物科技有限公司 | Preparation method of high-water-solubility rice protein |
| CN114431331B (en) * | 2021-12-31 | 2024-03-15 | 无锡金农生物科技有限公司 | Preparation method of rice protein with high water solubility |
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