CN107488688A - A kind of method for improving xylanase hydrolysis efficiency - Google Patents
A kind of method for improving xylanase hydrolysis efficiency Download PDFInfo
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- CN107488688A CN107488688A CN201710954398.6A CN201710954398A CN107488688A CN 107488688 A CN107488688 A CN 107488688A CN 201710954398 A CN201710954398 A CN 201710954398A CN 107488688 A CN107488688 A CN 107488688A
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- hydrolysis efficiency
- improving
- xylanase hydrolysis
- xylan
- zytase
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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
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Abstract
The invention provides a kind of method for improving xylanase hydrolysis efficiency.Comprise the following steps：1) xylan or pretreated lignocellulose raw material are scattered in pH=4.5 5.5 aqueous solution；2) zytase and nonionic surfactant/bovine serum albumin mixed liquor are added in above-mentioned mixed liquor, enzyme digestion reaction is carried out at 45 55 DEG C.It is an advantage of the invention that nonionic surfactant or bovine serum albumin are added during xylanase hydrolysis xylan either lignocellulose raw material, the vigor of zytase can be improved, so as to improve the hydrolysis efficiency of zytase, reduce the dosage of zytase, the cost of lignocellulose raw material production fermentable sugars is reduced, it is finally reduced and prepares the cost of bioenergy and chemicals.
Description
Technical field
The invention belongs to the technical field of biomass economy, specifically, relate to a kind of xylanase hydrolysis that improve and imitates
The method of rate.
Background technology
Using the lignocellulose raw material production bio-ethanol rich in cellulose and hemicellulose, the liquid fuel such as butanol for
Solves environmental pollution, reduction is of great immediate significance to the increasingly dependence of fossil fuel of scarcity etc. such as oil, coal.So
And the production process efficiency of bio-ethanol and butanol is low at present, cost is higher, these seriously limit bio-ethanol and butanol
Industrialized production.In lignocellulose raw material, hemicellulose occupies the 20-30% of its total amount, hydrolysis of hemicellulose generation
Alcohol fuel and butanol can be converted into after monose is fermented, so as to can greatly improve its yield.Thus, make full use of in substrate
Hemicellulose be one of key factor of biological liquid fuel economic feasibility.
But pass through hydrogen bond, ester between the three big component-celluloses, hemicellulose and lignin in lignocellulose raw material
Key, carbon-carbon key, acetal bonds, ehter bond, glycosidic bond etc. is combined closely so that enzyme molecule is difficult to be contacted with hemicellulose, causes wood
Glycanase hydrolysis efficiency is low.Therefore, the hydrolysis efficiency for improving zytase is the key for making full use of hemicellulose in substrate
Technology.
The content of the invention
It is an object of the invention to for xylanase hydrolysis efficiency it is low the problem of, there is provided one kind effectively improves xylan
The method of enzyme hydrolysis efficiency, the final transformation efficiency for improving xylan.Simultaneously in lignocellulose raw material xylan hydrolysis dissolution
Also accessibility of the cellulase to cellulose can be increased, improve fermentable sugars concentration in the hydrolysis efficiency and hydrolyzate of cellulose,
So as to improve the yield of subsequent fermentation production bio-ethanol and butanol, later separation cost is reduced.
The technical solution of the present invention is to provide a kind of method for improving xylanase hydrolysis efficiency, including following step
Suddenly：
Step 1：Xylan or pretreated lignocellulosic material are scattered in the pH=4.5-5.5 aqueous solution
In；
Step 2：Zytase is added into the mixed liquor of step 1, adds nonionic surfactant or cow's serum
Mixed liquid of protein, enzyme digestion reaction is carried out at 45-55 DEG C.
Preferably, above-mentioned nonionic surfactant is tween or polyethylene glycol.
Preferably, above-mentioned polyethylene glycol is Macrogol 6000 or polyethylene glycol 2000.
Preferably, above-mentioned tween is Tween 80 or polysorbate40.
Preferably, the concentration of above-mentioned nonionic surfactant or bovine serum albumin in system is 0.5-5.0mg/mL.
Preferably, above-mentioned lignocellulosic material is maize straw, corncob, bagasse, straw, the plant of straw class dogstail section
Thing, poplar class broadleaf or dragon spruce class softwood.
Preferably, diluted acid, diluted alkaline chemical method and/or attrition grinding physical method and/or steam blasting, hydro-thermal thing are passed through
Physicochemical method preprocessing lignocellulose.
Preferably, above-mentioned xylan is xylan after purification.
Preferably, zytase dosage is 200-2000nkat/g substrates in step 2.
Preferably, enzyme digestion reaction pH is equal to 4.5-5.0, hydrolysis time 48 hours.
The beneficial effects of the invention are as follows：
1st, it is provided by the invention to reduce lignin to wood by adding nonionic surfactant or bovine serum albumin
The unproductive absorption of dextranase, the hydrolysis efficiency of xylan is set to improve 9-28%；
2nd, the present invention promotes wood in lignocellulose raw material poly- by adding nonionic surfactant or bovine serum albumin
The hydrolysis of sugar, while the hydrolysis dissolution of xylan also increases accessibility of the cellulase to cellulose, so as to improve fiber
The transformation efficiency of element；
3rd, method technique of the invention is simple and convenient to operate suitable for large-scale application.
Embodiment
The present invention is described in further detail with reference to specific embodiment, but it is not as a limitation of the invention.
Embodiment one
Weigh the purified xylans of 1g to be placed in triangular flask, add a certain amount of zytase and Tween 80, add water, make
Concentration of substrate is 2% (w/v), and wherein w is the quality of substrate, and v is the volume for adding water, and zytase concentration is 500nkat/g
Substrate, Tween 80 concentration are 1mg/mL, and control hydrolyzation system pH is equal to 4.5, and the temperature of regulating thermostatic oscillator is 45 DEG C, by three
Angle bottle, which is placed in constant temperature oscillator, to be vibrated, and is persistently hydrolyzed.After hydrolysis 48 hours, hydrolyzate is taken out, 10 are handled with boiling water bath
Minute makes zytase lose vigor, and is centrifuged after cooling, takes supernatant, contained using high-efficient liquid phase chromatogram technique analysis xylose therein
Amount.Analysis result shows that after adding Tween 80, the conversion ratio of xylan improves 9%.
Embodiment two
Weigh the maize straw after 1g pretreatment with agueous Ammonia to be placed in triangular flask, add a certain amount of zytase and cow's serum
Albumen, add water, make concentration of substrate be 2% (w/v), wherein w is the quality of substrate, and v is the volume for adding water, zytase concentration
For 1000nkat/g substrates, bovine serum albumin concentration is 2mg/mL, and control hydrolyzation system pH is equal to 5.0, regulating thermostatic oscillator
Temperature be 50 DEG C, triangular flask is placed in constant temperature oscillator and vibrated, is persistently hydrolyzed.After hydrolysis 48 hours, hydrolysis is taken out
Liquid, being handled 10 minutes with boiling water bath makes zytase lose vigor, and is centrifuged after cooling, takes supernatant, using high performance liquid chromatography
Method analyzes Xylose Content therein.Analysis result shows after adding bovine serum albumin that the conversion ratio of xylan improves 28.3%.
Embodiment three
Weigh the corncob after 1g dilute acid pretreatments to be placed in triangular flask, add a certain amount of zytase and polyethylene glycol
6000, add water, make concentration of substrate be 2% (w/v), wherein w is the quality of substrate, and v is the volume for adding water, zytase concentration
For 2000nkat/g substrates, Macrogol 6000 concentration is 1mg/mL, and control hydrolyzation system pH is equal to 5.0, regulating thermostatic vibration
The temperature of device is 45 DEG C, and triangular flask is placed in constant temperature oscillator and vibrated, is persistently hydrolyzed.After hydrolysis 48 hours, water is taken out
Liquid is solved, being handled 10 minutes with boiling water bath makes zytase lose vigor, and is centrifuged after cooling, takes supernatant, using high-efficient liquid phase color
Chromatography Xylose Content therein.Analysis result shows after adding Macrogol 6000 that the conversion ratio of xylan improves
18.1%.
Example IV
Weigh the pretreated poplar of 1g steam blastings to be placed in triangular flask, add a certain amount of zytase and poly- second two
Alcohol 2000, adds water, and it is 2% (w/v) to make concentration of substrate, and wherein w is the quality of substrate, and v is the volume for adding water, and zytase is dense
It is 2.5mg/mL to spend for 1000nkat/g substrates, Macrogol 6000 concentration, and control hydrolyzation system pH is equal to 4.5, regulating thermostatic
The temperature of oscillator is 50 DEG C, and triangular flask is placed in constant temperature oscillator and vibrated, is persistently hydrolyzed.After hydrolysis 48 hours, take
Go out hydrolyzate, being handled 10 minutes with boiling water bath makes zytase lose vigor, and after cooling, centrifugation, supernatant is taken, using efficient liquid
GC headspace analysis Xylose Content therein.Analysis result shows after adding polyethylene glycol 2000 that steam blasting is pretreated
The conversion ratio of xylan improves 21.1% in poplar.
Embodiment five
Weigh the dragon spruce after 1g dilute acid pretreatments to be placed in triangular flask, add a certain amount of zytase and polysorbate40, add
Water, it is 2% (w/v) to make concentration of substrate, and wherein w is the quality of substrate, and v is the volume for adding water, and zytase concentration is
1000nkat/g substrates, polysorbate40 concentration are 4mg/mL, and control hydrolyzation system pH is equal to 5.0, the temperature of regulating thermostatic oscillator
For 50 DEG C, triangular flask is placed in constant temperature oscillator and vibrated, is persistently hydrolyzed.After hydrolysis 48 hours, hydrolyzate is taken out, with boiling
Water bath processing makes zytase lose vigor in 10 minutes, is centrifuged after cooling, takes supernatant, using high-efficient liquid phase chromatogram technique analysis its
In Xylose Content.Analysis result shows after adding polysorbate40 that the conversion ratio of xylan improves 15.8% in dragon spruce.
Claims (10)
- A kind of 1. method for improving xylanase hydrolysis efficiency, it is characterised in that comprise the following steps：Step 1：Xylan or pretreated lignocellulosic material are scattered in the pH=4.5-5.5 aqueous solution；Step 2：Zytase is added into the mixed liquor of step 1, adds nonionic surfactant or bovine serum albumin Mixed liquor, enzyme digestion reaction is carried out at 45-55 DEG C.
- 2. the method according to claim 1 for improving xylanase hydrolysis efficiency, it is characterised in that：The non-ionic surface Activating agent is tween or polyethylene glycol.
- 3. the method according to claim 2 for improving xylanase hydrolysis efficiency, it is characterised in that：The polyethylene glycol is Macrogol 6000 or polyethylene glycol 2000.
- 4. the method according to claim 2 for improving xylanase hydrolysis efficiency, it is characterised in that：The tween is tween 80 or polysorbate40.
- 5. the method according to claim 2 for improving xylanase hydrolysis efficiency, it is characterised in that：The non-ionic surface The concentration of activating agent or bovine serum albumin in system is 0.5-5.0mg/mL.
- 6. according to the method for any described raising xylanase hydrolysis efficiency of claim 1-5, it is characterised in that：It is described wooden Cellulosic material is maize straw, corncob, bagasse, straw, straw class dogstail section plant, poplar class broadleaf or dragon spruce class Softwood.
- 7. the method according to claim 6 for improving xylanase hydrolysis efficiency, it is characterised in that：Pass through diluted acid, diluted alkaline Chemical method and/or attrition grinding physical method and/or steam blasting, hydro-thermal physico-chemical process preprocessing lignocellulose.
- 8. the method according to claim 7 for improving xylanase hydrolysis efficiency, it is characterised in that：The xylan is pure Xylan after change.
- 9. the method according to claim 8 for improving xylanase hydrolysis efficiency, it is characterised in that：Xylan in step 2 Enzyme dosage is 200-2000nkat/g substrates.
- 10. the method according to claim 9 for improving xylanase hydrolysis efficiency, it is characterised in that：Enzyme digestion reaction pH etc. In 4.5-5.0, hydrolysis time 48 hours.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109337947A (en) * | 2018-10-30 | 2019-02-15 | 华南农业大学 | A method of Heating Explosion Sugarcane Bagasse efficiency is improved by alkaline peroxide impregnation and additive |
| CN113881732A (en) * | 2021-11-01 | 2022-01-04 | 华南农业大学 | Enzymolysis accelerant for improving enzymolysis efficiency of cellulose and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101696427A (en) * | 2009-11-03 | 2010-04-21 | 天津科技大学 | Method for producing fuel ethanol and 2,3-butanediol by using fibrous matter |
-
2017
- 2017-10-13 CN CN201710954398.6A patent/CN107488688A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101696427A (en) * | 2009-11-03 | 2010-04-21 | 天津科技大学 | Method for producing fuel ethanol and 2,3-butanediol by using fibrous matter |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109337947A (en) * | 2018-10-30 | 2019-02-15 | 华南农业大学 | A method of Heating Explosion Sugarcane Bagasse efficiency is improved by alkaline peroxide impregnation and additive |
| CN113881732A (en) * | 2021-11-01 | 2022-01-04 | 华南农业大学 | Enzymolysis accelerant for improving enzymolysis efficiency of cellulose and application thereof |
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