CN105112474A - Application of tetraacetylethylenediamine to improvement of lignocellulose enzymolysis conversion rate - Google Patents
Application of tetraacetylethylenediamine to improvement of lignocellulose enzymolysis conversion rate Download PDFInfo
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- CN105112474A CN105112474A CN201510613208.5A CN201510613208A CN105112474A CN 105112474 A CN105112474 A CN 105112474A CN 201510613208 A CN201510613208 A CN 201510613208A CN 105112474 A CN105112474 A CN 105112474A
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Abstract
The invention relates to application of tetraacetylethylenediamine to improvement of the lignocellulose enzymolysis conversion rate, and aims to decompose hydrogen peroxide through a complete reaction to release active oxygen and improve the lignocellulose enzymolysis conversion rate. The application comprises the following steps: uniformly mixing tetraacetylethylenediamine (TAED), NaOH, H2O2 and deionized water, adding a pretreated lignocellulose raw material, and carrying out a 20-30 hours' heat preservation reaction, thereby obtaining treated lignocellulose. According to a treatment method related to the application, hydrogen peroxide is decomposed into hydrogen peroxide anions under an alkali condition, a nucleophilic substitution reaction can be carried out between TAED and the hydrogen peroxide anions to generate peroxy acetic acid anions and DAED, peroxy acetic acid can act on lignin more efficiently to activate H2O2, and lignin can be oxidized and separated out of cellulose through released active oxygen, so that the inhibition of lignin to follow-up enzymolysis is eliminated, and the glucose conversion rate is effectively improved.
Description
Technical field
The present invention relates to a kind of tetraacetyl ethylene diamine improving the application on lignocellulose enzymolysis transformation efficiency, belonging to biomass resource trans-utilization technical field.
Background technology
Lignocellulose is the renewable resources of the abundantest cheapness on the earth, accumulates and stores up huge biomass energy.Be mainly derived from agricultural wastes, forest waste and urban waste.Along with the exhaustion of fossil oil and going from bad to worse of environment, lignocellulose sugar is changed into glucose and xylose etc., and be further used for the production of bio-based chemical as alcohol fuel equal energy source product, be subject to growing interest.China is large agricultural country; agricultural straw resource is quite abundant; produce nearly 800,000,000 tons per year; but for a long time; major part stalk is dropped or burns; not only waste resource but also pollute environment, the right combination of stalk resource utilizes significant for protection of the environment, alleviating energy crisis and promotion agricultural sustainable development.
Utilize lignocellulose to produce important bio-based chemical and mainly comprise the steps such as pre-treatment, enzymolysis conversion and fermentation as alcohol fuel etc.Wherein preconditioning technique is the committed step that lignocellulose transforms, by destroying the connection between Cellulose-lignins-hemicellulose, reducing cellulosic degree of crystallinity, removing xylogen, to make cellulase and hemicellulase effectively contact Mierocrystalline cellulose and hemicellulose, thus improve enzymolysis efficiency.
But acid system preprocessing lignocellulose easily produces materials such as suppressing the furfural of microorganism growth and hydroxymethylfurfural; Vapour quick-fried method cost is high, and product is complicated, has tar formation.
Chinese patent literature CN102758028A (application number 201110104494.4) discloses a kind of preprocessing lignocellulose raw material, then is converted into the method for reducing sugar.It adopts liquid-solid ratio 5-50, and lignocellulose is processed 1-10 hour in the mixing solutions of the hydrogen peroxide of massfraction 0-5% sodium hydroxide and massfraction 0-5%, and after process, gained solid residue washes oven dry with water.Then at acidolysis time 10-120min, acidolysis temperature 30-150 DEG C, h 2 so 4 concentration 0-10%, liquid-solid ratio 5-50 process solid residue, adjust ph to 7, then dries.This treatment process can make xylogen effectively remove, and hemicellulose solves five-carbon sugar at diluted acid Water Under, through reacting further and can obtaining high value added product.
But aforesaid method is used in the preprocessing process of lignocellulose enzyme process hydrolysis, and enzymolysis transformation efficiency is relatively low, cannot meet related request.
Hydrogen peroxide is as normally used discoloring agent; other by products are not had to produce in addition to water; but simple hydrogen peroxide for decoloration effect is unsatisfactory; in order to improve drift colour efficiency; by research tetraacetyl ethylene diamine (TAED) as auxiliary agent; SYNTHETIC OPTICAL WHITNER (hydrogen peroxide) can be activated; hydrogen bond can be formed with hydrogen peroxide in hydrolytic process; transition state is stablized; and be easily hydrolyzed, do not form acyl peroxide, to improve the bleaching effect of hydrogen peroxide; but in the prior art, tetraacetyl ethylene diamine (TAED) Chang Zuowei bleaching assistant uses.
Summary of the invention
For the deficiencies in the prior art, the invention provides a kind of tetraacetyl ethylene diamine and improving the application on lignocellulose enzymolysis transformation efficiency.
Technical scheme of the present invention is as follows:
Tetraacetyl ethylene diamine is improving the application on lignocellulose enzymolysis transformation efficiency, decomposing and discharging active oxygen, improving lignocellulose enzymolysis transformation efficiency, comprising step as follows for making hydrogen peroxide complete reaction:
By tetraacetyl ethylene diamine, NaOH, H
2o
2mix with deionized water, add pretreated lignocellulosic material, obtain mixed reaction solution, then under temperature 28 ~ 35 DEG C, rotating speed 120 ~ 200rpm condition, insulation reaction 20 ~ 30 hours, solid-liquid separation after reaction, solid deionized water wash, drying, obtain the lignocellulose after process;
In described mixed reaction solution, the add-on of tetraacetyl ethylene diamine is the pretreated lignocellulosic material of 0.05 ~ 0.3g/g, and the add-on of NaOH is the pretreated lignocellulosic material of 0.1 ~ 0.5g/g, H
2o
2add-on be the pretreated lignocellulosic material of 0.05 ~ 0.3g/g, the add-on of deionized water is the pretreated lignocellulosic material of 40 ~ 60ml/g.
Preferred according to the present invention, the add-on of tetraacetyl ethylene diamine is the pretreated lignocellulosic material of 0.08 ~ 0.15g/g, and further preferably, the add-on of tetraacetyl ethylene diamine is the pretreated lignocellulosic material of 0.1g/g.
Preferred according to the present invention, the add-on of NaOH is the pretreated lignocellulosic material of 0.08 ~ 0.15g/g, and further preferably, the add-on of NaOH is the pretreated lignocellulosic material of 0.1 ~ 0.15g/g.
Preferred according to the present invention, H
2o
2add-on be the pretreated lignocellulosic material of 0.09 ~ 0.15g/g, further preferably, H
2o
2add-on be the pretreated lignocellulosic material of 0.1g/g.
Preferred according to the present invention, the add-on of deionized water is the pretreated lignocellulosic material of 50ml/g.
Preferred according to the present invention, pretreated lignocellulosic material is that foreign material removed by raw material, then by the sieve of lignocellulose through air-dry, pulverizing, excessively 5mm aperture, obtains pretreated lignocellulosic material.
Preferred according to the present invention, lignocellulosic material is selected from maize straw, straw, cotton stalk, straw, Rape Straw, sweet sorghum stalk, bagasse, wood chip, waste paper and/or switchgrass.
Preferred according to the present invention, first NaOH is added after dissolving in the deionized water of meltage, then add in system.
Utilize the lignocellulose after above-mentioned process to produce a method for xylose and glucose, comprise as follows:
A, in the lignocellulose after above-mentioned obtained process, add tsiklomitsin, cycloheximide and citrate buffer solution, the addition of tsiklomitsin reaches 35 ~ 45 μ g/mL for making tsiklomitsin final concentration, the addition of cycloheximide reaches 25 ~ 35 μ g/mL for making cycloheximide final concentration, citrate buffer solution addition reach 40 ~ 60ul/mL for making citrate buffer solution final concentration;
B, then add prozyme, the total dosage of zymoprotein is 40 ~ 60mg/g Mierocrystalline cellulose, and under 45 ~ 50 DEG C of conditions, stirring reaction 65 ~ 75 hours, through centrifugal, gets supernatant, obtained xylose and glucose solution.
The present invention is preferred, and the concentration of citrate buffer solution is 1mol/L, pH is 4.8.
The present invention is preferred, and described prozyme component is beta-glucosidase and cellulase Accelerase1000, and beta-glucosidase is 1:4 with the protein ratio of cellulase Accelerase1000.
Preferred according to the present invention, in described step b, stirring velocity is 100 ~ 200rpm, and centrifugal is at 4 DEG C, centrifugal 15min under the condition of 12000rpm.
The present inventor surprisingly finds, treatment process of the present invention, tetraacetyl ethylene diamine (TAED) and hydrogen peroxide acting in conjunction can improve lignocellulose enzymolysis transformation efficiency greatly, hydrogen peroxide can be decomposed into perhydroxy anion in the basic conditions, TAED energy and perhydroxy anion generation nucleophilic substitution reaction generate Peracetic Acid negatively charged ion and diacetyl ethylenediamine (DAED), and Peracetic Acid more effectively can act on xylogen, in enzymolysis process, the existence due to xylogen makes the enzymic hydrolysis efficiency of lignocellulose reduce.TAED of the present invention adds, and can make H
2o
2activated, discharging active oxygen can separate out lignin oxidation from Mierocrystalline cellulose, removes the inhibition of xylogen to enzymolysis thereafter, effectively improves the transformation efficiency of glucose.The addition of tetraacetyl ethylene diamine (TAED) is that those skilled in the art gropes to obtain through long-term experiment, the too small raising to transformation efficiency of addition of tetraacetyl ethylene diamine (TAED) does not have effect, the addition of tetraacetyl ethylene diamine (TAED) is crossed conference and is played lignocellulose hydrolytic action, finally impels transformation efficiency to improve.
Beneficial effect of the present invention is as follows:
Through treatment process of the present invention, tetraacetyl ethylene diamine and hydrogen peroxide acting in conjunction can improve lignocellulose enzymolysis transformation efficiency greatly, tetraacetyl ethylene diamine makes hydrogen peroxide complete reaction decompose and discharges active oxygen, improve lignocellulose enzymolysis transformation efficiency, cellulosic sugared transformation efficiency is impelled to improve further, this preprocessing process is simple, does not produce unmanageable waste water.
Accompanying drawing explanation
The treatment process that Fig. 1, embodiment 1 treatment process and comparative example 1 do not add tetraacetyl ethylene diamine affects histogram to the content of lignin after processing, and the add-on of NaOH is the pretreated lignocellulosic material of 0.1g/g;
The treatment process that Fig. 2, embodiment 2 treatment process and comparative example 2 do not add tetraacetyl ethylene diamine affects histogram to the content of lignin after processing, and the add-on of NaOH is the pretreated lignocellulosic material of 0.15g/g;
The histogram that the treatment process that Fig. 3, embodiment 1 treatment process and comparative example 1 do not add tetraacetyl ethylene diamine affects the transformation efficiency of glucose and xylose, the add-on of NaOH is the pretreated lignocellulosic material of 0.1g/g;
The histogram that the treatment process that Fig. 4, embodiment 2 treatment process and comparative example 2 do not add tetraacetyl ethylene diamine affects the transformation efficiency of glucose and xylose, the add-on of NaOH is the pretreated lignocellulosic material of 0.15g/g.
Embodiment
Below by specific embodiment, also the present invention will be further described by reference to the accompanying drawings, but be not limited thereto.
In embodiment, beta-glucosidase and cellulase Accelerase1000 are purchased from Novozymes Company;
Tetraacetyl ethylene diamine, commercial products, market is buied.
Embodiment 1
Tetraacetyl ethylene diamine is improving the application on lignocellulose enzymolysis transformation efficiency, and decompose for making hydrogen peroxide complete reaction and discharge active oxygen, improve lignocellulose enzymolysis transformation efficiency, embody rule step is as follows:
(1) remove the foreign material in maize straw, then lignocellulose is sieved through sieve that is air-dry, that pulverize, cross 5mm aperture, obtain pretreated lignocellulosic material;
(2) tetraacetyl ethylene diamine, NaOH, H
2o
2mix with deionized water, add pretreated lignocellulosic material, obtain mixed reaction solution, mixed reaction solution is under temperature 28 DEG C, rotating speed 150rpm condition, and insulation reaction 24 hours, reacts rear solid-liquid separation, solid deionized water wash, drying, obtain the lignocellulose after process;
In described mixed reaction solution, the add-on of tetraacetyl ethylene diamine is the pretreated lignocellulosic material of 0.1g/g, and the add-on of NaOH is the pretreated lignocellulosic material of 0.1g/g, H
2o
2add-on be the pretreated lignocellulosic material of 0.1g/g, the add-on of deionized water is the pretreated lignocellulosic material of 50ml/g.
Embodiment 2
Tetraacetyl ethylene diamine is improving the application on lignocellulose enzymolysis transformation efficiency, and decompose for making hydrogen peroxide complete reaction and discharge active oxygen, improve lignocellulose enzymolysis transformation efficiency, embody rule step is as follows:
(1) remove the foreign material in maize straw, then lignocellulose is sieved through sieve that is air-dry, that pulverize, cross 5mm aperture, obtain pretreated lignocellulosic material;
(2) by tetraacetyl ethylene diamine, NaOH, H
2o
2mix with deionized water, add pretreated lignocellulosic material, obtain mixed reaction solution, mixed reaction solution is under temperature 30 DEG C, rotating speed 150rpm condition, and insulation reaction 24 hours, reacts rear solid-liquid separation, solid deionized water wash, drying, obtain the lignocellulose after process;
In described mixed reaction solution, the add-on of diethylenetriamine pentamethylenophosphonic acid(DTPP) is the pretreated lignocellulosic material of 0.1g/g, and the add-on of NaOH is the pretreated lignocellulosic material of 0.15g/g, H
2o
2add-on be the pretreated lignocellulosic material of 0.15g/g, the add-on of deionized water is the pretreated lignocellulosic material of 50ml/g.
Comparative example 1
Experiment condition is with embodiment 1, and difference is, the method step (2) does not add tetraacetyl ethylene diamine, and the add-on of NaOH is the pretreated lignocellulosic material of 0.1g/g.
Comparative example 2
Experiment condition is with embodiment 1, and difference is, the method step (2) does not add tetraacetyl ethylene diamine, and the add-on of NaOH is the pretreated lignocellulosic material of 0.15g/g.
Test example 1
Utilize lignocellulose to produce a method for xylose and glucose, comprise as follows:
A, in the lignocellulose after the obtained process of embodiment 1 ~ 2, add tsiklomitsin, cycloheximide and concentration be 1mol/L, pH is the lemon acid buffer of 4.8, the addition of tsiklomitsin reaches 40 μ g/mL for making tsiklomitsin final concentration, the addition of cycloheximide reaches 30 μ g/mL for making cycloheximide final concentration, citrate buffer solution addition reach 50ul/mL for making citrate buffer solution final concentration;
B, then add prozyme, prozyme component is beta-glucosidase and cellulase Accelerase1000, beta-glucosidase is 1:4 with the protein ratio of cellulase Accelerase1000, the total dosage of zymoprotein is 50mg/g Mierocrystalline cellulose then shaking table 50 DEG C of constant temperature hydrolysis 72h under rotating speed is 150rpm condition, with 15min centrifugal under the condition of 12000rpm, get supernatant, obtained xylose and glucose solution.
Adopt pretreated lignocellulose prepared by comparative example 1, produce xylose and glucose solution according to the method described above, be designated as control group 1.
Adopt the pretreated lignocellulose that comparative example 2 is standby, produce xylose and glucose solution according to the method described above, be designated as control group 2.
According to the standard method (NREL/TP-510-42618 in NREL (National Renewable Energy laboratory); NREL/TP-510-42619; NREL/TP-510-42623) content of water content, Mierocrystalline cellulose, hemicellulose, acid-soluble xylogen, sour insoluble xylogen is detected.Utilize the glucose and xylose content in the above-mentioned obtained product of high effective liquid chromatography for measuring, according to following formulae discovery grape sugar and xylose rate:
Cellulose conversion rate=C
glucose* Mierocrystalline cellulose total amount before V*0.9/ hydrolysis
Hemicellulose transformation efficiency=C
wood sugarhemicellulose total amount before * 0.88/ hydrolysis
Content of lignin measures the standard method with reference to U.S. NREL: DeterminationofStructuralCarbohydratesandLignininBiomass
The histogram that embodiment 1, embodiment 2 process rear content of lignin is shown in shown in Fig. 1, Fig. 2.
By Fig. 1, Fig. 2, the content balance of xylogen after process, content of lignin is lower than comparative example 1, comparative example 2 after the disposal methods of the embodiment of the present invention 1, embodiment 2, by Fig. 3, Fig. 4, treatment process and the comparative example treatment process of the embodiment of the present invention 1, embodiment 2 contrast the transformation efficiency of glucose and xylose, treatment process of the present invention is obviously better than comparative example 1 improving the transformation efficiency of glucose, synthesizing map 1 to 4, the existence due to xylogen makes the enzymic hydrolysis efficiency of lignocellulose reduce.Treatment process of the present invention more effectively can act on xylogen, removes the inhibition of xylogen to enzymolysis thereafter more, effectively improves the transformation efficiency of glucose.Effectively improve the transformation efficiency of glucose.
Claims (10)
1. tetraacetyl ethylene diamine is improving the application on lignocellulose enzymolysis transformation efficiency, decomposing and discharging active oxygen, improving lignocellulose enzymolysis transformation efficiency, comprising step as follows for making hydrogen peroxide complete reaction:
By tetraacetyl ethylene diamine, NaOH, H
2o
2mix with deionized water, add pretreated lignocellulosic material, obtain mixed reaction solution, then under temperature 28 ~ 35 DEG C, rotating speed 120 ~ 200rpm condition, insulation reaction 20 ~ 30 hours, solid-liquid separation after reaction, solid deionized water wash, drying, obtain the lignocellulose after process;
In described mixed reaction solution, the add-on of tetraacetyl ethylene diamine is the pretreated lignocellulosic material of 0.05 ~ 0.3g/g, and the add-on of NaOH is the pretreated lignocellulosic material of 0.1 ~ 0.5g/g, H
2o
2add-on be the pretreated lignocellulosic material of 0.05 ~ 0.3g/g, the add-on of deionized water is the pretreated lignocellulosic material of 40 ~ 60ml/g.
2. tetraacetyl ethylene diamine according to claim 1 is improving the application on lignocellulose enzymolysis transformation efficiency, it is characterized in that, the add-on of tetraacetyl ethylene diamine is the pretreated lignocellulosic material of 0.08 ~ 0.15g/g, preferably, the add-on of tetraacetyl ethylene diamine is the pretreated lignocellulosic material of 0.1g/g.
3. tetraacetyl ethylene diamine according to claim 1 is improving the application on lignocellulose enzymolysis transformation efficiency, it is characterized in that, the add-on of NaOH is the pretreated lignocellulosic material of 0.08 ~ 0.15g/g, preferably, the add-on of NaOH is the pretreated lignocellulosic material of 0.1 ~ 0.15g/g.
4. tetraacetyl ethylene diamine according to claim 1 is improving the application on lignocellulose enzymolysis transformation efficiency, it is characterized in that, H
2o
2add-on be the pretreated lignocellulosic material of 0.09 ~ 0.15g/g, preferably, the add-on of NaOH is the pretreated lignocellulosic material of 0.1g/g.
5. tetraacetyl ethylene diamine according to claim 1 is improving the application on lignocellulose enzymolysis transformation efficiency, and it is characterized in that, the add-on of deionized water is the pretreated lignocellulosic material of 50ml/g.
6. tetraacetyl ethylene diamine according to claim 1 is improving the application on lignocellulose enzymolysis transformation efficiency, it is characterized in that, pretreated lignocellulosic material is that foreign material removed by raw material, then by the sieve of lignocellulose through air-dry, pulverizing, excessively 5mm aperture, pretreated lignocellulosic material is obtained.
7. tetraacetyl ethylene diamine according to claim 1 is improving the application on lignocellulose enzymolysis transformation efficiency, it is characterized in that, lignocellulosic material is selected from maize straw, straw, cotton stalk, straw, Rape Straw, sweet sorghum stalk, bagasse, wood chip, waste paper and/or switchgrass.
8. the lignocellulose after utilizing claim 1 to process produces a method for xylose and glucose, comprises as follows:
A, in the lignocellulose after above-mentioned obtained process, add tsiklomitsin, cycloheximide and citrate buffer solution, the addition of tsiklomitsin reaches 35 ~ 45 μ g/mL for making tsiklomitsin final concentration, the addition of cycloheximide reaches 25 ~ 35 μ g/mL for making cycloheximide final concentration, citrate buffer solution addition reach 40 ~ 60ul/mL for making citrate buffer solution final concentration;
B, then add prozyme, the total dosage of zymoprotein is 40 ~ 60mg/g Mierocrystalline cellulose, and under 45 ~ 50 DEG C of conditions, stirring reaction 65 ~ 75 hours, through centrifugal, gets supernatant, obtained xylose and glucose solution.
9. the lignocellulose after utilizing process according to Claim 8 produces the method for xylose and glucose, and it is characterized in that, the concentration of citrate buffer solution is 1mol/L, pH is 4.8.
10. the lignocellulose after utilizing process according to Claim 8 produces the method for xylose and glucose, it is characterized in that, described prozyme component is beta-glucosidase and cellulase Accelerase1000, beta-glucosidase is 1:4 with the protein ratio of cellulase Accelerase1000, in described step b, stirring velocity is 100 ~ 200rpm, and centrifugal is at 4 DEG C, centrifugal 15min under the condition of 12000rpm.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102758028A (en) * | 2011-04-26 | 2012-10-31 | 中国石油大学(北京) | Method for pre-treating lignocellulose raw material and for preparing reducing sugar by conversion |
| CN104404108A (en) * | 2014-12-09 | 2015-03-11 | 齐鲁工业大学 | Pre-treating method for improving sugar conversion rate of lignocellulose |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102758028A (en) * | 2011-04-26 | 2012-10-31 | 中国石油大学(北京) | Method for pre-treating lignocellulose raw material and for preparing reducing sugar by conversion |
| CN104404108A (en) * | 2014-12-09 | 2015-03-11 | 齐鲁工业大学 | Pre-treating method for improving sugar conversion rate of lignocellulose |
Non-Patent Citations (3)
| Title |
|---|
| 张晓阳等: "《纤维素生物质水解与应用》", 31 December 2012, 郑州大学出版社 * |
| 贺近恪等: "《林产化学工业全书》", 28 February 2001, 中国林业出版社 * |
| 钱学仁等: "四乙酰乙二胺对麦草浆H2O2漂白的活化作用", 《中国造纸》 * |
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Application publication date: 20151202 |