CN108893507A - A method of it is pre-processed by combination and promotes sawdust enzymatic hydrolysis - Google Patents
A method of it is pre-processed by combination and promotes sawdust enzymatic hydrolysis Download PDFInfo
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Abstract
The invention discloses the methods for promoting sawdust enzymatic hydrolysis by combination pretreatment, carry out efficient combination pretreatment to poplar sawdust using sour processing method combination alkali treatment method.This method specifically includes：(1) using poplar sawdust as raw material, xylo-oligosaccharide is prepared in acid processing；(2) after the acid processing poplar sawdust generated again using step (1) carries out oxygenation pretreatment as raw material, pretreatment poplar wood fiber element enzyme hydrolysis yield is greatly improved.In described (2) alkali treatment, Polyethylene Glycol Bisglycidyl Ether aid in treatment is added, lignin may advantageously facilitate the progress of cellulose hydrolyzation to the non-specific adsorption of cellulase during effectively weakening enzyme hydrolysis.Test result shows that compared with existing result of study, the enzyme hydrolysis yield of this patent significantly improves, and reaches 83.48%.Meanwhile the method for the present invention realizes the coproduction of xylo-oligosaccharide, the comprehensive utilization for poplar sawdust provides new way.
Description
Technical field
The present invention relates to sawdust pretreatment and enzymatic hydrolysis, and in particular to a kind of that the side for promoting sawdust enzymatic hydrolysis is pre-processed by combination
Method.
Background technique
As human society is to the growing and exhaustive exploitation of fossil resource demand, fossil resource is inevitable on the earth
Ground is faced with exhausted destiny totally.Lignocellulose raw material is renewable resource the most abundant on the earth, it passes through plant
Solar energy is captured and is stored in polymer form by photosynthesis.Lignocellulose raw material is turned using industrial biotechnology
Turning to fungible energy source and chemicals etc. is one of the important channel for alleviating fossil resource shortage.Lignocellulose raw material mainly includes
Broadleaf, softwood, energy crop, agricultural crop straw, agriculture and forestry organic waste material and city fiber rubbish etc..Poplar is that China is important
Economic tree, formed the complete industrial chain from breeding, cultivation, processing and utilization and Furniture manufacturing at present.According to incompletely statistics,
Up to 8,000,000 hectares or more, cultivated area and accumulation occupy first place in the world China's poplar artificial forest cultivated area.Thus band
The fast development of the China Dong Liao poplar secondary industry, therefore the poplar processing sawdust total amount generated is also extremely considerable.However, at present
These poplar sawdust utilization rates are lower, are mainly used for farmers' family's dyestuff, small part is for particieboard production, pulping and paper-making and food
With bacterium base-material.Therefore, under the overall background of energy restriction and environmental pollution, using poplar sawdust as raw material, pass through industrial bio skill
Art, is translated into the energy or chemicals has great importance.
Cellulose in poplar sawdust can be biodegradable into monosaccharide under cellulase catalytic, is then fermented by different microorganisms
The various energy or chemicals.However, as other lignocellulose raw materials, cellulose in poplar sawdust, hemicellulose and wooden
Element forms fine and close structure, is unfavorable for effect of the cellulase to cellulose.Therefore, lignocellulose raw material is carrying out cellulase
It must be pre-processed before hydrolysis, so that the compact texture of lignocellulosic is destroyed, to achieve the purpose that improve enzyme hydrolysis efficiency.
Common preprocess method has dilute acid pretreatment, steam blasting pretreatment, alkaline process pretreatment, hydrothermal pretreatment etc..It is pre- in diluted acid
In treatment process, hemicellulose is largely hydrolyzed in raw material, so that cellulose accessibility improves, greatly strengthens raw cellulose enzyme
Hydrolysis property.Alkaline process processing is then effectively broken ester bond or ehter bond, to be conducive to the dissolution of lignin.However, variety classes are wooden
The structural difference of matter fibrous raw material causes same preprocess method often to show different pretreating effects.Early-stage study
The result shows that herbaceous material is handled using conventional diluted acid method, such as corn stover, it can be achieved that the efficient fibre of pretreatment corn stover
Tie up plain enzyme hydrolysis.However, it is directed to woody raw material, even if improving dilute acid pretreatment intensity, dilute acid pretreatment poplar sawdust
Cellulose hydrolyzation yield is still in reduced levels.Equally, early-stage study the result shows that, polyethyleneglycol diglycidylether auxiliary
Low temperature alkali process corn stover, so that Spruce lignin realizes that lignin situ-formed graft is modified during oxygenation pretreatment,
To reduce lignin hydrophobicity, alleviates lignin to the ineffective adsorption of cellulase, significantly improve cellulose hydrolyzation yield.
However, the low temperature alkali process of polyethyleneglycol diglycidylether auxiliary can not abolish compact texture for poplar sawdust.If mentioned
High pretreatment temperature or pretreatment base amount, then often make under the rate of recovery of cellulose or hemicellulose is significant in woody raw material
Drop.It therefore, is the cellulose hydrolyzation performance for improving poplar sawdust, the high-efficiency pretreatment strategy for establishing poplar sawdust becomes urgently
It solves the problems, such as.
Summary of the invention
Goal of the invention：In order to solve the problems, such as that existing sawdust pretreating effect is poor, enzymatic hydrolysis yield is low, the present invention provides one
The method that kind promotes sawdust to digest by combination pretreatment.
Technical solution：A kind of method pre-processing promotion sawdust enzymatic hydrolysis by combination of the present invention, includes the following steps：
(1) low-kappa number is carried out to sawdust using acidic aqueous solution, it is water-soluble that separation of solid and liquid obtains the acidity containing xylo-oligosaccharide
Liquid and filter residue；
(2) oxygenation pretreatment is carried out to the filter residue that step (1) obtains using alkaline aqueous solution, is added in the alkaline aqueous solution
Polyethylene Glycol Bisglycidyl Ether；It is separated by solid-liquid separation after alkali process and obtains rich cellulose-containing material；
(3) the cellulose-containing material of richness obtained using cellulase degradation step (2), obtains glucose.
Preferably, the sawdust is poplar sawdust.
In step (1), the acidic aqueous solution is the aqueous acetic acid that volumetric concentration is 5-10% or mass concentration is
The solid-to-liquid ratio of the aqueous sulfuric acid or aqueous hydrochloric acid solution of 0.1-2%, sawdust and acidic aqueous solution is 1g:5-20mL (press by solid-to-liquid ratio
Sawdust over dry restatement, sawdust water content is low, and contained humidity is not counted in acidic aqueous solution), the condition of the low-kappa number is as follows：
140-190 DEG C of processing 0.4-1.0h.It is preferred that being used for subsequent alkali process after filter residue is washed, is filtered.
In step (2), the alkaline aqueous solution is the sodium hydrate aqueous solution of 10-50g/L, the filter residue and sodium hydroxide
The solid-to-liquid ratio of aqueous solution is 1g:(solid-to-liquid ratio presses filter residue over dry restatement to 5-20mL, and filter residue contained humidity is included in sodium hydrate aqueous solution
In, for calculating the concentration and volume of sodium hydrate aqueous solution), the condition of the oxygenation pretreatment is as follows：60-90 DEG C of processing 1-
4h；It is preferred that the concentration of sodium hydrate aqueous solution is 30-50g/L.The Polyethylene Glycol Bisglycidyl Ether is polyethyleneglycol shrink
The combination of any one or two kinds in glycerin ether, polyethyleneglycol diglycidylether.The Polyethylene Glycol Bisglycidyl Ether
Quality is the 5-30% of the filter residue quality, and preferably 20-30%, filter residue quality is by over dry restatement.It is preferred that will be enriched in cellulose
Material is washed, is filtered, and the lignin and sugar of remaining lye, dissolution are washed away.
In step (3), enzymatic hydrolysis system includes following component：The cellulose-containing object of richness that cellulase, step (2) obtain
Material, citrate buffer solution and water, cellulase degradation cellulose obtain glucose；The wherein moisture in rich cellulose-containing material
It is included in enzymatic hydrolysis system.The concentration of cellulose is 10-100g/L in the enzymatic hydrolysis system, and the dosage of cellulase is 10-40FPU/g
Cellulose.The enzymatic hydrolysis system digests 48-72h under the conditions of 45-55 DEG C, pH 4.0-6.0.The enzyme activity of its cellulase is fixed
Justice is as follows：The international unit (FPU) of one filter paper enzyme activity is equal to generates 1 μm of ol glucose per minute under the conditions of standard reaction
The enzyme amount of amount.
Beneficial effect：Compared with prior art, the efficient combination preprocess method of poplar sawdust of the invention, at acetic acid
After reason, it is added the oxygenation pretreatment of Polyethylene Glycol Bisglycidyl Ether, lignin is to fiber during effectively reducing enzyme hydrolysis
The non-specific adsorption of plain enzyme may advantageously facilitate the progress of cellulose hydrolyzation.Acetic acid processing has effectively hydrolyzed in lignin
Aromatic yl ether key provides more to release more free phenolic hydroxyl groups for the grafting and modifying of Polyethylene Glycol Bisglycidyl Ether
Reaction site.This effectively improves the Hydrophilic modification degree of lignin, further enhances lignin in-situ modification pair
The relaxation effect of enzyme non-specific adsorption.Test result shows and individually uses acetic acid processing or Polyethylene Glycol Bisglycidyl Ether
The enzymatic hydrolysis yield of auxiliary oxygenation pretreatment poplar sawdust is compared, and combination pretreatment of the invention is so that enzyme hydrolysis yield is improved by 30%
To 80% or more.The achievable xylo-oligosaccharide coproduction of the present invention, the comprehensive utilization for poplar sawdust each component provide one simultaneously
New way.
Detailed description of the invention
Fig. 1 is zymoprotein adsorption rate result figure after combined treatment poplar sawdust enzyme hydrolysis 72h.
Specific embodiment
In following embodiment, glucose and xylose concentration is measured using high performance liquid chromatography (HPLC).Chromatographic condition is such as
Under：Chromatograph：Agillent1200 high performance liquid chromatograph；Chromatographic column：Bio-Rad Aminex HPX-87H；Mobile phase：
0.005mol/L sulfuric acid, flow velocity：0.6mL/min；Column temperature：55℃；Detector：Differential refraction detector；Sample volume：10μL.Outside
The measurement of mark method.
In following embodiment, xylo-oligosaccharide concentration is measured using indirect acid hydrolyzation in acetic acid treatment fluid.Specific measuring method
It is as follows：10mL acetic acid treatment fluid is taken, pH is adjusted to 7.0 or so with sodium hydroxide, and be settled to 50mL；Taking 10mL, treated
10mL 8% (w/v) sulfuric acid is added in acidolysis bottle in sample solution, handles 60min at 121 DEG C.Acid is measured using HPLC
Xylose concentration after solution in sample and in acetic acid treatment fluid, and xylo-oligosaccharide concentration is calculated by xylose concentration difference before and after acidolysis.
Xylose concentration g/L in xylose concentration g/L × 2 × 5- acetic acid treatment fluid in sample after xylo-oligosaccharide concentration (g/L)=acidolysis.Formula
In：2 and 5 be extension rate.
In embodiment, polyethyleneglycol diglycidylether and polyethylene glycol monoglycidyl ether dosage are with acetic acid processing poplar
Sawdust over dry restatement refers to pretreatment 100g over dry acetic acid processing by taking polyethyleneglycol diglycidylether dosage 20% (g/g) as an example
Poplar sawdust, polyethyleneglycol diglycidylether dosage are 20g, and polyethyleneglycol diglycidylether molecular weight is 500.
Embodiment 1
A kind of efficient combination preprocess method of poplar sawdust, steps are as follows：
(1) weighing over dry weight is 200g poplar sawdust, with 5% (v/v) aqueous acetic acid, in solid-to-liquid ratio 1:10 (g/mL, under
Together), 0.5h is handled at 170 DEG C.
(2) acetic acid after treatment is separated by solid-liquid separation, and the treatment fluid of generation is that the acetic acid rich in xylo-oligosaccharide is water-soluble
Liquid, xylo-oligosaccharide concentration and yield are shown in Table 1；Its generate solid residue washed, filter after obtain acetic acid processing poplar wood
Bits, Principle components analysis are shown in Table 1.
Wherein, xylo-oligosaccharide yield (%)=(1000g × xylan contains (concentration of oligosaccharide g/L × 10 in hydrolyzate) ÷
Amount) × 100%.In formula：1 handles total volume, L for acetic acid；1000 be substrate over dry weight, g.
(3) weighing over dry weight is that the above-mentioned acetic acid of 20g handles poplar sawdust, with 10-50g/L sodium hydrate aqueous solution, poly- second
Hexanediol diglycidyl ether dosage 20%, solid-to-liquid ratio 1:10g/mL, coprocessing 3h at 70 DEG C.Wherein, polyethylene glycol diglycidyl is sweet
Oily ether quality handles poplar sawdust over dry restatement by acetic acid.
(4) after combined treatment, material is washed and is collected by filtration.At polyethyleneglycol diglycidylether Assisted Combinatorial
The Principle components analysis of reason poplar sawdust is shown in Table 2.
(5) said combination that cellulose over dry weight is 1.00g is weighed respectively handles poplar sawdust in 250mL hydrolysis bottle,
1mol/L citrate buffer solution 2.5mL, enzyme dosage is added as the cellulase of 20FPU/g cellulose in each hydrolysis bottle and fits
Distilled water is measured, makes moisture total volume 50mL in enzymatic hydrolysis system, closes the lid after being mixed well reaction system with glass rod, in
72h is digested in the constant-temperature table that 4.8,50 DEG C of 150rpm, pH.
After enzymatic hydrolysis, respectively hydrolysate is centrifuged 5min with centrifuge under the conditions of 10000rpm, supernatant is taken to measure
Concentration of glucose therein, and enzyme hydrolysis yield is calculated, data take double parallel to test average value.Wherein, cellulase hydrolysis yield
(%)=(concentration of glucose g/L × 0.9 in hydrolyzate) × 0.05 ÷ (substrate over dry weight g × beta-dextran content) × 100%.Formula
In：0.9 is the conversion coefficient of glucose and cellulose；0.95 is the conversion coefficient of cellobiose and cellulose；0.05 is hydrolysis
Liquid product, L.
Acetic acid processing poplar sawdust Principle components analysis and xylo-oligosaccharide yield are shown in Table 1.
1 acetic acid of table handles poplar sawdust Principle components analysis and xylo-oligosaccharide yield
Table 1 the result shows that, acetic acid is handled so that xylan hydrolysis is monosaccharide and xylo-oligosaccharide in poplar sawdust, and xylan contains
Amount is remarkably decreased, and glucan and the insoluble content of lignin of acid significantly increase.In addition, detecting oligomeric wood in acetic acid treatment fluid
Sugar, the content based on xylan in untreated poplar sawdust calculate its xylo-oligosaccharide yield up to 37.60%.
It adds the combined treatment poplar sawdust Principle components analysis of 20% polyethyleneglycol diglycidylether and digests
Rate is shown in Table 2.
2 polyethyleneglycol diglycidylether Assisted Combinatorial of table handles the Principle components analysis and enzymatic hydrolysis yield of poplar sawdust
Table 2 the result shows that, under identical polyethyleneglycol diglycidylether dosage (20%, g/g), with pretreatment alkali use
The increase of amount, beta-dextran content gradually increases in combined treatment poplar sawdust, and xylan and the insoluble content of lignin of acid are gradually
It reduces, while cellulase hydrolysis yield significantly improves.
Embodiment 2
A kind of efficient combination preprocess method of poplar sawdust, steps are as follows：
(1) weighing over dry weight is 200g poplar sawdust, with 5% (v/v) aqueous acetic acid, in solid-to-liquid ratio 1:10 (g/mL, under
Together), 0.5h is handled at 170 DEG C.
(2) acetic acid after treatment is separated by solid-liquid separation, and collects treatment fluid and processing solid residue respectively；Solid residue
It is stand-by after being washed, being filtered.
(3) weighing over dry weight is that the above-mentioned acetic acid of 20g handles poplar sawdust, with 30g/L sodium hydrate aqueous solution, polyethylene glycol
Diglycidyl ether dosage is 5-30%, solid-to-liquid ratio 1:10g/mL, coprocessing 3h at 70 DEG C.Wherein, polyethylene glycol diglycidyl is sweet
Oily ether quality handles poplar sawdust over dry restatement by acetic acid.
(4) after combined treatment, material is washed and is collected by filtration.At polyethyleneglycol diglycidylether Assisted Combinatorial
The Principle components analysis of reason poplar sawdust is shown in Table 3.
(5) said combination that cellulose over dry weight is 1.00g is weighed respectively handles poplar sawdust in 250mL hydrolysis bottle,
1mol/L citrate buffer solution 2.5mL, enzyme dosage is added as the cellulase of 20FPU/g cellulose in each hydrolysis bottle and fits
Distilled water is measured, makes moisture total volume 50mL in enzymatic hydrolysis system, closes the lid after being mixed well reaction system with glass rod, in
72h is digested in the constant-temperature table that 4.8,50 DEG C of 150rpm, pH.
After enzymatic hydrolysis, respectively hydrolysate is centrifuged 5min with centrifuge under the conditions of 10000rpm, supernatant is taken to measure
Concentration of glucose therein, and enzyme hydrolysis yield is calculated, data take double parallel to test average value.Polyethyleneglycol diglycidylether
Assisted Combinatorial processing poplar sawdust 72h enzymatic hydrolysis yield is shown in Table 3.
3 polyethyleneglycol diglycidylether Assisted Combinatorial of table handles the Principle components analysis and enzymatic hydrolysis yield of poplar sawdust
From table 3 the result shows that, under identical alkali concentration (30g/L), with the increasing of polyethyleneglycol diglycidylether dosage
Add, pre-processes the insoluble content of lignin of acid in corn stover and gradually increase, pretreated material enzymatic hydrolysis yield gradually rises.When poly- second
When Hexanediol diglycidyl ether dosage is more than 20%, the raising trend for digesting yield tends towards stability.
Embodiment 3
A kind of efficient combination preprocess method of poplar sawdust, cellulase adsorb situation test, and specific step is as follows：
(1) weighing over dry weight is 200g poplar sawdust, with 5% (v/v) aqueous acetic acid, in solid-to-liquid ratio 1:10 (g/mL, under
Together), 0.5h is handled at 170 DEG C.
(2) acetic acid after treatment is separated by solid-liquid separation, and collects treatment fluid and processing solid residue respectively；Solid residue
It is stand-by after being washed, being filtered.
(3) weighing over dry weight is that the above-mentioned acetic acid of 20g handles poplar sawdust, with 30g/L sodium hydrate aqueous solution, polyethylene glycol
Diglycidyl ether dosage is 5-30%, solid-to-liquid ratio 1:10g/mL, coprocessing 3h at 70 DEG C.
(4) after combined treatment, material is washed and is collected by filtration, and is operated for cellulase hydrolysis.
(5) said combination that cellulose over dry weight is 1.00g is weighed respectively handles poplar sawdust in 250mL hydrolysis bottle,
1mol/L citrate buffer solution 2.5mL, enzyme dosage is added as the cellulase of 20FPU/g cellulose in each hydrolysis bottle and fits
Distilled water is measured, makes moisture total volume 50mL in enzymatic hydrolysis system, closes the lid after being mixed well reaction system with glass rod, in
72h is digested in the constant-temperature table that 4.8,50 DEG C of 150rpm, pH.
(6) after digesting 72h, respectively hydrolysate is centrifuged 5min with centrifuge under the conditions of 10000rpm, supernatant is taken to survey
The concentration of the fixed zymoprotein that wherein dissociates, and opposite protein content is calculated, data take double parallel to test average value.Wherein, enzyme egg
White adsorption rate (%)=(zymoprotein concentration g/L in the total zymoprotein concentration g/L- supernatant added in enzymatic hydrolysis system) ÷ (enzymatic hydrolysis
The total zymoprotein concentration g/L added in system) × 100%.
To compare with polyethyleneglycol diglycidylether is not added in combined treatment, test is synchronized.As a result such as Fig. 1
It is shown, in the poplar sawdust enzymatic hydrolysis check experiment for being not added with polyethyleneglycol diglycidylether combined treatment, digest group after 72h
It closes processing poplar sawdust and the 84.8% of total protein content is reached to cellulase adsorbance.However, in the proteolysis assay of test sample
In, as polyethyleneglycol diglycidylether additive amount increases, digest suction of the combined treatment poplar sawdust to cellulase after 72h
Attached rate gradually decreases.When polyethyleneglycol diglycidylether additive amount is 30%, 72h zymoprotein adsorption rate drops to
56.0%, it is far below control group.Illustrate that adding polyethyleneglycol diglycidylether Assisted Combinatorial processing poplar sawdust reduces object
Expect the non-specific adsorption to cellulase.
Embodiment 4
A kind of efficient combination preprocess method of poplar sawdust, the measurement of lignin phenol's hydroxy radical content, specific step is as follows：
(1) weighing over dry weight is 200g poplar sawdust, with 5% (v/v) aqueous acetic acid, in solid-to-liquid ratio 1:10 (g/mL, under
Together), 0.5h is handled at 170 DEG C；Acetic acid after treatment, is separated by solid-liquid separation, and collects treatment fluid and processing solid residue respectively；
Solid residue is stand-by after being washed, being filtered, and is denoted as material 1.
(2) weighing over dry weight is 20g poplar sawdust, with 30g/L sodium hydrate aqueous solution, solid-to-liquid ratio 1:10g/mL, 70 DEG C
Lower coprocessing 3h；After treatment, material are washed and are collected by filtration for use, and material 2 is denoted as.
(3) weighing over dry weight is that the above-mentioned acetic acid of 20g handles poplar sawdust (material 1), with 30g/L sodium hydrate aqueous solution,
Solid-to-liquid ratio 1:10g/mL, coprocessing 3h at 70 DEG C；After the combined treatment for being not added with polyethyleneglycol diglycidylether, material
It is washed and is collected by filtration, meter does material 3.
(4) weighing over dry weight is that the above-mentioned acetic acid of 20g handles poplar sawdust (material 1), with 30g/L sodium hydrate aqueous solution,
Polyethyleneglycol diglycidylether dosage is 20%, solid-to-liquid ratio 1:10g/mL, coprocessing 3h at 70 DEG C；After combined treatment,
Material is washed and is collected by filtration, and meter does material 4.
(5) above-mentioned material 1-4 is taken, after drying process, carries out ball-milling treatment in ball mill, ball milling speed 600rpm,
Effective Ball-milling Time is 6h.
(6) take above-mentioned ball milling wood powder in dioxane/water (96:4, v/v) lignin extraction is carried out in solvent, solid-to-liquid ratio is
It is 1:10, extracting gained lignin is Milled wood lignin.
(7) it uses31P nuclear magnetic resonance technique analyzes the content of phenolic hydroxyl groups of above-mentioned 4 kinds of Milled wood lignins, and the results are shown in Table 4.
Lignin phenol's hydroxy radical content (mmol/g) in 4 four kinds of pretreatment poplar sawdusts of table
| Material 1 | Material 2 | Material 3 | Material 4 | |
| Content of phenolic hydroxyl groups | 1.79 | 1.01 | 2.07 | 0.12 |
The result shows that, individually used in low temperature alkali process poplar sawdust (material 2), the free phenol of lignin from table 4
Hydroxy radical content is minimum (1.01mmol/g).Polyethyleneglycol diglycidylether auxiliary is individually handled and is not added with using acetic acid
After combined treatment, the free phenolic hydroxyl group content of lignin is significantly increased in pretreatment poplar sawdust (material 1 and material 3).Explanation
Acetic acid processing has effectively hydrolyzed the aromatic yl ether key in lignin, is polyethylene glycol diglycidyl to release free phenolic hydroxyl group
The grafting and modifying of glycerin ether provides more reaction sites.In addition, in the group of addition polyethyleneglycol diglycidylether auxiliary
It closes in processing poplar sawdust (material 4), the free phenolic hydroxyl group content of lignin is remarkably decreased to 0.12mmol/g, is further illustrated
Polyethyleneglycol diglycidylether successfully grafts on lignin phenol's hydroxyl.
Embodiment 5
For method with embodiment 1, different places is as follows：
The volumetric concentration of aqueous acetic acid is 10% in step (1), and the solid-to-liquid ratio of poplar sawdust and aqueous acetic acid is 1:5
(g/mL), 140 DEG C of processing 1.0h；
The solid-to-liquid ratio of filter residue and sodium hydrate aqueous solution is 1 in step (3):5 (g/mL), addition are polyethyleneglycol contracting
Water glycerin ether, 60 DEG C of processing 4h；
The combination poplar sawdust that cellulose over dry weight is 5.00g is weighed in step (5), the dosage of cellulase is 40FPU/
G cellulose, 55 DEG C of enzymatic hydrolysis 72h.
Embodiment 6
For method with embodiment 1, different places is as follows：
The volumetric concentration of aqueous acetic acid is 10% in step (1), and the solid-to-liquid ratio of poplar sawdust and aqueous acetic acid is 1:
20 (g/mL), 190 DEG C of processing 0.4h；
The solid-to-liquid ratio of filter residue and sodium hydrate aqueous solution is 1 in step (3):20 (g/mL), 90 DEG C of processing 1h；
The combination poplar sawdust that cellulose over dry weight is 0.5g is weighed in step (5), the dosage of cellulase is 10FPU/g
Cellulose, 45 DEG C of enzymatic hydrolysis 48h.
Comparative example 1
A kind of poplar sawdust acetic acid processing method individually pre-processes poplar sawdust using acetic acid, without alkali
Processing, step does not include step (3) and (4) with the step (1), (2) and (5) in embodiment 2.Wherein, step (2) obtains
The Principle components analysis and enzymatic hydrolysis yield of acetic acid processing poplar sawdust are shown in Table 5.
5 acetic acid of table handles the Principle components analysis and enzymatic hydrolysis yield of poplar sawdust
Table 5 the result shows that, individually using acetic acid handle poplar sawdust, enzymatic hydrolysis yield be only 32.31%, far below implement
The combined treatment poplar sawdust that polyethyleneglycol diglycidylether auxiliary is added in example 1 and embodiment 2 digests yield (52.63-
83.48%).
Comparative example 2
A kind of poplar sawdust polyethyleneglycol diglycidylether auxiliary alkali treatment method, i.e., individually contracted using polyethylene glycol two
Water glycerin ether auxiliary alkaline process pre-processes poplar sawdust, and specific step is as follows：
(1) weighing over dry weight is 20g poplar sawdust, and with 30-50g/L sodium hydrate aqueous solution, polyethylene glycol diglycidyl is sweet
Oily ether dosage is 20%, solid-to-liquid ratio 1:10g/mL, coprocessing 3h at 70 DEG C.
(2) after treatment, material are washed and are collected by filtration；It adds polyethyleneglycol diglycidylether and assists alkali process
The Principle components analysis of poplar sawdust is shown in Table 6.
(3) said combination that cellulose over dry weight is 1.00g is weighed respectively handles poplar sawdust in 250mL hydrolysis bottle,
1mol/L citrate buffer solution 2.5mL, enzyme dosage is added as the cellulase of 20FPU/g cellulose in each hydrolysis bottle and fits
Distilled water is measured, makes moisture total volume 50mL in enzymatic hydrolysis system, closes the lid after being mixed well reaction system with glass rod, in
72h is digested in the constant-temperature table that 4.8,50 DEG C of 150rpm, pH.
After enzymatic hydrolysis, respectively hydrolysate is centrifuged 5min with centrifuge under the conditions of 10000rpm, supernatant is taken to measure
Concentration of glucose therein, and enzyme hydrolysis yield is calculated, data take double parallel to test average value, the results are shown in Table 6.
Table 6 adds the Principle components analysis and enzymatic hydrolysis yield of polyethyleneglycol diglycidylether auxiliary alkali process poplar sawdust
From table 6 the result shows that, under identical polyethyleneglycol diglycidylether dosage (20%, g/g), when oxygenation pretreatment alkali
When concentration is 30-50g/L, enzymatic hydrolysis yield is only 30.13-35.32%, is equally far below in embodiment 1 and embodiment 2 and adds
The combined treatment poplar sawdust of polyethyleneglycol diglycidylether auxiliary digests yield (52.63-83.48%).
Comparative example 3
A kind of efficient combination preprocess method of poplar sawdust, step do not add poly- second in alkali process with embodiment 1
Hexanediol diglycidyl ether is not added with the Principle components analysis of polyethyleneglycol diglycidylether Assisted Combinatorial processing poplar sawdust
It the results are shown in Table 7 with enzymatic hydrolysis.
Table 7 is not added with the Principle components analysis and enzymatic hydrolysis of polyethyleneglycol diglycidylether Assisted Combinatorial processing poplar sawdust
Yield
Table 7 the result shows that, compared with the result of embodiment 1, under identical base amount, the insoluble lignin of acid in embodiment 1
Content (table 2) is above the insoluble content of lignin of acid in the present embodiment.This is polyethyleneglycol diglycidylether grafting and modifying wood
The result of quality.In addition, the enzymatic hydrolysis yield of the present embodiment is below the enzymatic hydrolysis of embodiment 1 under identical alkali concentration treatment conditions
Yield, when handling alkali concentration is 30g/L, the enzymatic hydrolysis yield of embodiment 1 is 80.86% (table 1), than digesting for the present embodiment
Rate (37.89%) improves 2.13 times.
Claims (10)
1. a kind of pre-process the method for promoting sawdust enzymatic hydrolysis by combination, which is characterized in that include the following steps：
(1) low-kappa number is carried out to sawdust using acidic aqueous solution, be separated by solid-liquid separation obtain acidic aqueous solution containing xylo-oligosaccharide and
Filter residue；
(2) oxygenation pretreatment is carried out to the filter residue that step (1) obtains using alkaline aqueous solution, adds poly- second in the alkaline aqueous solution
Glycol glycidol ether；It is separated by solid-liquid separation after alkali process and obtains rich cellulose-containing material；
(3) the cellulose-containing material of richness obtained using cellulase degradation step (2), obtains glucose.
2. the method according to claim 1, wherein sawdust described in step (1) is poplar sawdust.
3. the method according to claim 1, wherein acidic aqueous solution described in step (1) is that volumetric concentration is
The aqueous acetic acid or mass concentration of 5-10% is the aqueous sulfuric acid or aqueous hydrochloric acid solution of 0.1-2%, and sawdust and acidity are water-soluble
The solid-to-liquid ratio of liquid is 1g:The condition of 5-20mL, the low-kappa number are as follows：140-190 DEG C of processing 0.4-1.0h, sawdust quality are pressed
Sawdust over dry restatement.
4. the method according to claim 1, wherein alkaline aqueous solution described in step (2) is 10-50g/L's
The solid-to-liquid ratio of sodium hydrate aqueous solution, the filter residue and sodium hydrate aqueous solution is 1g:5-20mL, the condition of the oxygenation pretreatment
It is as follows：60-90 DEG C of processing 1-4h, filter residue quality press filter residue over dry restatement.
5. according to the method described in claim 4, it is characterized in that, the concentration of the sodium hydrate aqueous solution is 30-50g/L.
6. the method according to claim 1, wherein Polyethylene Glycol Bisglycidyl Ether described in step (2) is poly-
The combination of any one or two kinds in ethylene glycol monoglycidyl ether, polyethyleneglycol diglycidylether.
7. the method according to claim 1, wherein the matter of Polyethylene Glycol Bisglycidyl Ether described in step (2)
Amount is the 5-30% of the filter residue quality, and filter residue quality presses over dry restatement.
8. method described in -7 any one according to claim 1, which is characterized in that enzymatic hydrolysis system includes as follows in step (3)
Component：The cellulose-containing material of richness, citrate buffer solution and the water that cellulase, step (2) obtain, cellulase degradation fiber
Element obtains glucose.
9. according to the method described in claim 8, it is characterized in that, the concentration of cellulose is 10-100g/ in the enzymatic hydrolysis system
L, the dosage of cellulase are 10-40FPU/g cellulose.
10. according to the method described in claim 8, it is characterized in that, the enzymatic hydrolysis system at 45-55 DEG C, pH 4.0-6.0 item
48-72h is digested under part.
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| CN110714041A (en) * | 2019-11-19 | 2020-01-21 | 云南省烟草农业科学研究院 | Method for efficiently and directly producing glucose by using waste tobacco stems |
| CN111349671A (en) * | 2020-02-21 | 2020-06-30 | 南京林业大学 | Method for promoting wood chip enzymolysis through combined pretreatment |
| CN113481260A (en) * | 2021-07-23 | 2021-10-08 | 南京林业大学 | Method for combined pretreatment of woody raw materials and co-production of xylo-oligosaccharide and glucose |
| CN114317638A (en) * | 2022-01-18 | 2022-04-12 | 中南林业科技大学 | Method for saccharifying lignocellulose through cyclic enzymolysis of multiple enzymes and surfactant |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110714041A (en) * | 2019-11-19 | 2020-01-21 | 云南省烟草农业科学研究院 | Method for efficiently and directly producing glucose by using waste tobacco stems |
| CN111349671A (en) * | 2020-02-21 | 2020-06-30 | 南京林业大学 | Method for promoting wood chip enzymolysis through combined pretreatment |
| CN113481260A (en) * | 2021-07-23 | 2021-10-08 | 南京林业大学 | Method for combined pretreatment of woody raw materials and co-production of xylo-oligosaccharide and glucose |
| CN114317638A (en) * | 2022-01-18 | 2022-04-12 | 中南林业科技大学 | Method for saccharifying lignocellulose through cyclic enzymolysis of multiple enzymes and surfactant |
| CN114317638B (en) * | 2022-01-18 | 2023-09-22 | 中南林业科技大学 | Method for saccharifying lignocellulose by multi-enzyme synergistic surfactant cyclic enzymolysis |
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