CN107164424A - It is a kind of to aoxidize the lignocellulose pretreatment method that delignification improves enzyme hydrolysis rate - Google Patents
It is a kind of to aoxidize the lignocellulose pretreatment method that delignification improves enzyme hydrolysis rate Download PDFInfo
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- CN107164424A CN107164424A CN201710583436.1A CN201710583436A CN107164424A CN 107164424 A CN107164424 A CN 107164424A CN 201710583436 A CN201710583436 A CN 201710583436A CN 107164424 A CN107164424 A CN 107164424A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/02—Monosaccharides
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P2201/00—Pretreatment of cellulosic or lignocellulosic material for subsequent enzymatic treatment or hydrolysis
Abstract
The method that delignification improves the lignocellulosic pretreatment of enzyme hydrolysis rate is aoxidized the present invention relates to a kind of, this method is comprised the following steps that：Lignocellulosic material is crushed, dried, controls SODIUM PERCARBONATE concentration, solid-to-liquid ratio, pretreatment temperature and time in suitable scope respectively, filtering is washed to neutrality, and pretreated lignocellulosic residue is obtained after drying；The cellulase and zytase that~5.0 citrate buffer solution dissolvings are added through pH4.5 into residue obtained above are digested, the fermentable reduction sugar juice of acquisition.Preprocess method in the present invention can effectively remove lignin, and cellulose and hemi-cellulose components retention rate are high, are effectively improved the enzymolysis efficiency and Reducing sugar of lignocellulosic, while also having environment-friendly, simple to operate, low cost and other advantages.
Description
Technical field
The invention belongs to lignocellulose biomass application technology as the second resource field, and in particular to one kind uses strong oxidizer
Delignification is aoxidized to improve the preprocess method of lignocellulose biomass enzyme hydrolysis rate.
Background technology
In the case where facing the dual-pressure of economic development and environmental protection, it is the world to develop the renewable resources such as biomass
Various countries set up the effective measures of sustainable energy system.Lignocellulose-like biomass is most huge, most potential in nature
Renewable resource, its effective trans-utilization has immeasurable value to human kind sustainable development.At present, by wooden fibre
Dimension element is hydrolyzed to reduced sugar by bioanalysis and obtains required product as lignocellulosic sourcesization profit through microbial fermentation again
With the study hotspot and main direction of studying in field.
The main component of lignocellulosic is cellulose (35-50%), hemicellulose (20-40%) and lignin (15-
25%).Lignocellulosic it is complicated, hemicellulose and lignin in cell membrane by covalent bond formation network structure,
Cellulose is inlayed wherein, and this complicated network structure is the natural anti-degraded barrier in biomass transformation process.It is especially wooden
Plain composition hinders hydrolase to enter biomass contact carbohydrate in the enzymolysis process of lignocellulosic, and can adsorb water
Solution enzyme influences its hydrolysing activity, causes enzymolysis efficiency to reduce, so as to increase the conversion cost of biorefining process.Therefore, go
Lignin is to release the natural anti-degraded barrier of lignocellulosic, improve the top priority of glycoside hydrolase functioning efficiency.
At present, conventional lignocellulose pretreatment method (ionic liquid method, acid-base pretreatment method, steam explosion, ozone
Method for pretreating etc.) in, the preprocess method of alkaline reagent and strong oxidizer combination is reported the wood that can effectively remove in biomass
Quality, while the hydrogen bond and the crystal structure of cellulose that destroy between hemicellulose, cellulose, so as to destroy stupid in biomass
Resistive connection structure, but research papers are less.SODIUM PERCARBONATE is also referred to as sodium carbonate peroxide or sodium carbonate peroxyhydrate, and its aqueous solution is in
Alkalescence, is a kind of alkaline strong oxidizer, is usually used in bleaching agent, detergent, fruit antistaling agent, oxygen-making agent metal conditioner
Deng, safe, cost is low, at home and abroad in enterprise extensively production and apply.
The content of the invention
Strong oxidizer SODIUM PERCARBONATE preprocessing lignocellulose and follow-up enzyme are used it is an object of the invention to provide one kind
The method of solution, the present invention can effectively remove the anti-degraded barrier in lignin, destruction lignocellulosic enzymolysis process, and then improve
The enzyme hydrolysis efficiency of lignocellulose biomass.Specifically include following steps：
(1) it is lignocellulosic material is (any in maize straw, corncob, wheat stalk, rice straw and bagasse
A kind of or its combination) crush, 40~80 mesh sieves are crossed after 90~105 DEG C of drying.
(2) raw material of a certain amount of size-reduced drying is taken, adding SODIUM PERCARBONATE solution makes solid-to-liquid ratio control 1: 5~1: 25,
It is preferred with 1: 10；Make the concentration of SODIUM PERCARBONATE control to be 1~8% (w/v), be preferred with 4%；Pretreatment temperature is controlled 30~70
DEG C, it is preferred with 60 DEG C；Pretreatment time is controlled in 0.5~8h, is preferred with 4h.
(3) after lignocellulosic residue filtering after pretreatment, with distillation water washing until filtrate pH reaches neutrality, 90
~105 DEG C of dryings are to constant weight.
(4) residue obtained through above-mentioned pretreatment is added pH in 4.5~5.0 citrate buffer solutions, to control its concentration
For 5~15% (w/v), it is preferred with 8%.
(5) the plain enzyme of commercial fibers and zytase from trichoderma are added into above-mentioned system, enzyme is controlled respectively
Useful load is 20~50FPU/g substrates (being preferred with 30FPU/g substrates) and 5000~12000U/g substrates (with 8000U/g substrates
It is preferred), 45~55 DEG C, 48~96h of reaction is digested under conditions of 100~200rpm of shaking speed, with 50 DEG C, shaking speed
150rpm, enzymolysis 72h is preferred.
(6) reduced sugar and glucose, xylose concentration in hydrolyzate are determined with DNS methods and HPLC methods respectively.
The present invention has the advantage that：The present invention is using strong oxidizer SODIUM PERCARBONATE processing wood safe, with low cost
Matter cellulosic material, can be achieved the clean and effective separation of lignin, and the lignocellulosic surface after being handled through SODIUM PERCARBONATE is fluffy
It is porous, by lignocellulosic microstructure analysis before and after the processing, as a result show connecting key hydrogen bond, ehter bond and the ester between three kinds of components
Key greatly reduces, and after illustrating the preprocess method processing through the present invention, the micro-structural of lignocellulosic material is destroyed, and is led to
The constituent content analysis to raw material before and after pretreatment is crossed, shows lignin removing rate up to 50%, cellulose and hemi-cellulose components
Retention rate is up to more than 90%.Enzymolysis experiment shows that the inventive method is greatly improved the enzymatic hydrolyzation of lignocellulosic, Portugal
Grape sugar yield is more than doubled, and xylose yield improves more than 2 times.The inventive method is simple to operation, safety non-pollution,
With feasible industrial applications prospect.
Embodiment
Below by embodiment, the present invention will be further described, and following embodiments are illustrative, be not it is limited,
Protection scope of the present invention can not be limited with following embodiments.
Embodiment 1
It is a kind of to aoxidize the preprocess method that delignification improves corn stalk raw material enzyme hydrolysis rate, comprise the following steps：
(1) by corn straw smashing, 60 mesh sieves are crossed after 90 DEG C of drying to constant weight.
(2) corn stalk raw material of the size-reduced drying of 10g is taken, 100mL concentration is added molten for 4% (w/v) SODIUM PERCARBONATE
Liquid, 4h is handled at 60 DEG C.
(3) pretreated maize straw is filtered, with distillation water washing until filtrate pH reaches neutrality, 90 DEG C of drying are extremely
Constant weight.
(4) (or pretreated maize straw is not former for the maize straw residue for taking after the above-mentioned pretreatments of 2.0g through drying
Material), 25mL 0.05mol/L, pH is added in 4.8 citrate buffer solution, the commodity of Trichoderma source are added into the mixed liquor
Cellulose enzyme and zytase, the useful load of its cellulase is 30FPU/g substrates, and the useful load of zytase is
8000U/g substrates, it is 40mL to control hydrolyzation system cumulative volume.
(5) above-mentioned system is digested under conditions of 50 DEG C, shaking speed 150rpm, is determined respectively also after reaction 72h
Raw sugar, glucose and xylose concentration, the results are shown in Table 1.
Table 1 pre-processes front and rear maize straw component analysis and the sugared yield of enzymolysis
Embodiment 2
It is a kind of to aoxidize the preprocess method that delignification improves corncob dhdps enzyme percent hydrolysis, comprise the following steps：
(1) corn core raw material is crushed, crosses 80 mesh sieves after 90 DEG C of drying to constant weight.
(2) the corn core raw material of the size-reduced drying of 10g is taken, the SODIUM PERCARBONATE solution that 100mL concentration is 5% (w/v) is added,
6h is handled at 50 DEG C.
(3) pretreated corncob is filtered, with distillation water washing until filtrate pH reaches neutrality, 90 DEG C of drying to perseverance
Weight.
(4) the corncob residue (or not pretreated corn core raw material) after the above-mentioned pretreatments of 2.0g through drying is taken, plus
Enter in the citrate buffer solution that 25mL 0.05mol/L, pH are 4.8, the commercial fibers of Trichoderma source are added into the mixed liquor
Plain enzyme and zytase, the useful load of its cellulase is 40FPU/g substrates, and the useful load of zytase is 10000U/g bottoms
Thing, it is 40mL to control hydrolyzation system cumulative volume.
(5) above-mentioned system is digested under conditions of 50 DEG C, shaking speed 180rpm, is determined respectively also after reaction 72h
Raw sugar, glucose and xylose concentration, the results are shown in Table 2.
Table 2 pre-processes front and rear corncob component analysis and the sugared yield of enzymolysis
Claims (6)
1. a kind of aoxidize the lignocellulose pretreatment method that delignification improves enzyme hydrolysis rate, it is characterised in that including following
Step：
(1) lignocellulosic material is crushed, dried, SODIUM PERCARBONATE concentration, solid-to-liquid ratio, pretreatment temperature and time are controlled respectively
In in suitable scope, filter, washing obtains pretreated lignocellulosic residue after drying；
(2) cellulase dissolved through citrate buffer solution is added into residue obtained above and zytase is digested,
Determine concentration of reduced sugar in hydrolyzate.
2. according to the method described in claim 1, it is characterised in that described lignocellulosic is maize straw, corncob, small
Any one of Wheat Straw, rice straw and bagasse or its combination.
3. according to the method described in claim 1, it is characterised in that after lignocellulosic material is crushed in step (1), cross 40~
80 mesh sieves, 90~105 DEG C of dryings to constant weight.
4. according to the method described in claim 1, it is characterised in that the concentration of SODIUM PERCARBONATE used is 1~8% in step (1)
(w/v)；The solid-to-liquid ratio that lignocellulosic material is mixed with SODIUM PERCARBONATE solution is controlled 1: 5~1: 25；Pretreatment temperature is controlled
At 30~70 DEG C；Pretreatment time is controlled in 0.5~8h；Lignocellulosic residue after pretreatment is filtered, distilled water is used
Washing is until filtrate pH reaches neutrality, 90~105 DEG C of dryings to constant weight.
5. according to the method described in claim 1, it is characterised in that the pH of citrate buffer solution described in step (2) is 4.5~
5.0；Cellulase and zytase are from trichoderma commercialization enzyme preparation.
6. according to the method described in claim 1, it is characterised in that in step (2) concentration of lignocellulosic residue be 5~
15% (w/v)；The useful load of lignocellulosic residue enzymatic hydrolysis system cellulase is 20~50FPU/g substrates, zytase
Useful load be 5000~12000U/g substrates；Enzymolysis is anti-under conditions of 45~55 DEG C of temperature, 100~200rpm of shaking speed
Answer 48~96h.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107630056A (en) * | 2017-11-15 | 2018-01-26 | 沈阳金博地生态环保科技有限公司 | A kind of method using immobilized bi-enzyme pretreated straw |
| CN108300747A (en) * | 2018-02-06 | 2018-07-20 | 西北农林科技大学 | A method of it is pre-processed using sodium chlorite and improves reed enzymatic saccharification efficiency |
| CN108588144A (en) * | 2018-03-29 | 2018-09-28 | 中国科学院广州能源研究所 | A method of preparing xylo-oligosaccharide and fermentable sugars using lignocellulose-like biomass |
| CN108659235A (en) * | 2018-04-12 | 2018-10-16 | 北京林业大学 | A kind of macromolecule polysaccharide prodegradant and the preparation method and application thereof |
| CN111593082A (en) * | 2020-07-03 | 2020-08-28 | 浙江华康药业股份有限公司 | Method for stabilizing lignocellulose enzymolysis process |
| CN111733197A (en) * | 2020-06-30 | 2020-10-02 | 南京林业大学 | Method for enzymatic hydrolysis and saccharification of plant fiber raw material |
| CN112553269A (en) * | 2020-12-24 | 2021-03-26 | 安徽工业大学 | Pretreatment and enzymolysis method of corn straw |
| CN112708647A (en) * | 2020-12-24 | 2021-04-27 | 安徽工业大学 | Method for improving enzymolysis and xylose conversion efficiency of corn straws by supercritical carbon dioxide coupled NaOH pretreatment |
| CN112852900A (en) * | 2019-11-27 | 2021-05-28 | 中国石油天然气股份有限公司 | Pretreatment method and enzymolysis method of lignocellulose raw material |
| 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 (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107630056A (en) * | 2017-11-15 | 2018-01-26 | 沈阳金博地生态环保科技有限公司 | A kind of method using immobilized bi-enzyme pretreated straw |
| CN108300747B (en) * | 2018-02-06 | 2020-11-27 | 西北农林科技大学 | Method for improving saccharified efficiency of silvergrass enzymolysis by utilizing sodium chlorite pretreatment |
| CN108300747A (en) * | 2018-02-06 | 2018-07-20 | 西北农林科技大学 | A method of it is pre-processed using sodium chlorite and improves reed enzymatic saccharification efficiency |
| CN108588144A (en) * | 2018-03-29 | 2018-09-28 | 中国科学院广州能源研究所 | A method of preparing xylo-oligosaccharide and fermentable sugars using lignocellulose-like biomass |
| CN108659235A (en) * | 2018-04-12 | 2018-10-16 | 北京林业大学 | A kind of macromolecule polysaccharide prodegradant and the preparation method and application thereof |
| CN108659235B (en) * | 2018-04-12 | 2020-04-14 | 北京林业大学 | Macromolecular polysaccharide degradation auxiliary agent and preparation method and application thereof |
| CN112852900A (en) * | 2019-11-27 | 2021-05-28 | 中国石油天然气股份有限公司 | Pretreatment method and enzymolysis method of lignocellulose raw material |
| CN111733197A (en) * | 2020-06-30 | 2020-10-02 | 南京林业大学 | Method for enzymatic hydrolysis and saccharification of plant fiber raw material |
| CN111593082A (en) * | 2020-07-03 | 2020-08-28 | 浙江华康药业股份有限公司 | Method for stabilizing lignocellulose enzymolysis process |
| CN111593082B (en) * | 2020-07-03 | 2023-10-13 | 浙江华康药业股份有限公司 | Method for stabilizing lignocellulose enzymolysis process |
| CN112553269A (en) * | 2020-12-24 | 2021-03-26 | 安徽工业大学 | Pretreatment and enzymolysis method of corn straw |
| CN112708647A (en) * | 2020-12-24 | 2021-04-27 | 安徽工业大学 | Method for improving enzymolysis and xylose conversion efficiency of corn straws by supercritical carbon dioxide coupled NaOH pretreatment |
| 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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