CN108588144A - A method of preparing xylo-oligosaccharide and fermentable sugars using lignocellulose-like biomass - Google Patents
A method of preparing xylo-oligosaccharide and fermentable sugars using lignocellulose-like biomass Download PDFInfo
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- CN108588144A CN108588144A CN201810274186.8A CN201810274186A CN108588144A CN 108588144 A CN108588144 A CN 108588144A CN 201810274186 A CN201810274186 A CN 201810274186A CN 108588144 A CN108588144 A CN 108588144A
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- oligosaccharide
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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
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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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
- C12P19/12—Disaccharides
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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 invention discloses a kind of methods preparing xylo-oligosaccharide and fermentable sugars using lignocellulose-like biomass, the lignin under alkaline condition in oxidation pre-treatment selectively removing lignocellulose-like biomass is used first, then xylo-oligosaccharide is obtained using the endo-xylanase hydrolysis pretreatment substrate without containing cellulase activity, the solid residue after the hydrolysis of cellulose hydrolyzation endo-xylanase is finally used to obtain fermentable sugars, this method not only eliminate high energy consumption cause it is of high cost, chemical levels lead to greatly the xylan separation and Extraction process of pollution weight, and also improve xylo-oligosaccharide purity and fermentable sugars yield.
Description
Technical field：
The present invention relates to technical field of biochemical industry, and in particular to it is a kind of using lignocellulose-like biomass prepare it is oligomeric
The method of xylose and fermentable sugars.
Background technology：
Xylo-oligosaccharide is the low molecule wood oligose for having special physiological function, because it can promote lactic acid bacteria, Bifidobacterium etc.
The proliferation of beneficial bacterium inhibits the growth of the pathogens such as clostridium, Escherichia coli, has and adjusts intestinal flora balance and change
The function of kind enteron aisle.Therefore xylo-oligosaccharide is acknowledged as a kind of good prebiotics, have higher market value.Xylo-oligosaccharide
The raw material of production is mainly the lignocellulose-like biomass such as corncob, bagasse and stalk.Currently, preparing the normal of xylo-oligosaccharide
Rule method needs to separate xylan from lignocellulosic, then zytase is recycled to be hydrolyzed to obtain oligomeric wood
Sugar, wherein high-temperature steam extract and concentrated base extracting is common xylan separation method.High-temperature steam extraction technique needs to use
Need high temperature resistant and high voltage bearing equipment, energy consumption larger.In addition, product is complex in high-temperature steam extracting processing procedure, second
Acid, furfural, acid-soluble lignin, pigment and the ash impurity that grades are more, lead to subsequently to refine that difficulty is big, and production cost is high.Concentrated base
Extracting needs to use a large amount of chemical reagent, and generates reluctant black liquor, in addition, in xylan purification process, needs
Using a large amount of acid neutralize and largely ethyl alcohol progress washing of precipitate could obtain the xylan substrate for enzyme hydrolysis.
The fermentable sugars such as the glucose and xylose that lignocellulosic hydrolysis generates can produce a large amount of combustion through microbial fermentation
Material and chemicals.Lignocellulosic is mainly made of cellulose, hemicellulose and three big component of lignin, and wherein lignin is formed
Physical chemistry barrier be to limit lignocellulosic enzyme hydrolysis to prepare the main bottleneck of xylo-oligosaccharide and fermentable sugars.Sodium chlorite
Although method can selectively remove lignin and retain cellulose and hemicellulose, sodium chlorite reagent environmental pollution compared with
Greatly, large-scale industrial production should not be carried out.In addition, xylan and its hydrolysis intermediate product can hinder and inhibit cellulase pair
The hydrolysis of cellulose.
Invention content：
The purpose of the present invention is to solve the above-mentioned problems of the prior art, provides and a kind of utilizing lignocellulose
The method that biomass prepares xylo-oligosaccharide and fermentable sugars uses oxidation pre-treatment selectively removing under alkaline condition wooden first
Lignin in cellulose series biomass, then using the endo-xylanase hydrolysis pretreatment bottom without containing cellulase activity
Object obtains xylo-oligosaccharide, and the solid residue after the hydrolysis of cellulose hydrolyzation endo-xylanase is finally used to obtain fermentable sugars,
This method, which not only eliminates high energy consumption, leads to of high cost, chemical levels greatly and causes the xylan separation and Extraction mistake of pollution weight
Journey, and also improve xylo-oligosaccharide purity and fermentable sugars yield.
The present invention is achieved by the following technical programs：
A method of xylo-oligosaccharide and fermentable sugars being prepared using lignocellulose-like biomass, this method includes following
Step：
(1) lignin selectively removing：The lignocellulose-like biomass crushed after drying is placed in glass reactor,
The lye that mass fraction is 0.5%~5% and the hydrogen peroxide that mass fraction is 0.5%~5% is added, control solid-liquid mass ratio is
1:10~35, reaction temperature is 30 DEG C~90 DEG C, and the reaction time is 1h~for 24 hours, is separated by solid-liquid separation after reaction, and be washed with water
Solid residue is to neutrality；
(2) prepared by xylo-oligosaccharide：With the solid residue obtained by pH5.0 buffer solutions dispersion steps (1), every gram of butt is then pressed
The endo-xylanase of 100~1000IU is added in substrate, reacts 6 under the conditions of 45~55 DEG C~carries out enzyme hydrolysis for 24 hours, reaction knot
It is separated by solid-liquid separation after beam, obtains solid residue after xylo-oligosaccharide solution and endo-xylanase hydrolysis；
(3) prepared by fermentable sugars：With the solid residue obtained by pH5.0 buffer solutions dispersion steps (2), every gram of butt is then pressed
10~50IU cellulases are added in substrate, and 48~96h is reacted under the conditions of 45~55 DEG C and carries out enzyme hydrolysis, after reaction solid-liquid
Separation, obtains fermentable sugars solution.
Preferably, the lignocellulose-like biomass is in bagasse, wheat stalk, maize straw or corncob
It is a kind of.
Preferably, the one kind of alkali in sodium hydroxide, potassium hydroxide or ammonium hydroxide in the step (1).
Preferably, the one kind of endo-xylanase in glycoside hydrolase Families 10 or 11 in the step (2).
Compared with prior art, the present invention having the advantages that：
1. the lignin under alkaline condition of the present invention in oxidation pre-treatment selectively removing lignocellulose-like biomass, so
Endo-xylanase direct hydrolysis is used to pre-process substrate afterwards, environmental-friendly, mild condition, not only eliminating high energy consumption causes into
This height, chemical levels lead to greatly xylan extraction and the separation process of pollution weight, and improve xylo-oligosaccharide purity.
2. the present invention using the solid residue after the hydrolysis of cellulose hydrolyzation endo-xylanase, removes lignin and wood is poly-
The non-cellulose component such as sugar contributes to the hydrolysis efficiency for increasing cellulase to cellulose, to improve cellulose hydrolyzation effect
Rate and inversion rate of glucose improve fermentable sugars yield.
Specific implementation mode：
It is the further explanation to the present invention below, rather than limiting the invention.
Embodiment 1：
By crush dry after bagasse be placed in glass reactor, be added mass fraction be 0.5% sodium hydroxide and
The hydrogen peroxide that mass fraction is 1%, solid-to-liquid ratio 1:10(g:G), reaction temperature is 70 DEG C, and reaction time 4h is used after filtering
Water washing solid residue obtains pretreatment substrate to neutrality.With the pretreatment substrate of pH5 buffer solutions dispersion gained, then press every
10 family's endo-xylanase of glycoside hydrolase of 300IU is added in gram butt substrate, and progress enzyme water for 24 hours is reacted under the conditions of 45 DEG C
Solid residue after xylo-oligosaccharide solution and endo-xylanase hydrolysis is obtained by filtration in solution.Wood with pH5 buffer solutions dispersion gained is poly-
Then solid residue after carbohydrase hydrolysis presses every gram of butt substrate and 15IU cellulases is added, 72h is reacted under the conditions of 50 DEG C and is carried out
Fermentable sugars solution is obtained by filtration in enzyme hydrolysis.Show that xylo-oligosaccharide conversion ratio is 21.12% by high performance liquid chromatography measurement,
Inversion rate of glucose is 81.25%, and xylose rate is 26.36%.
Embodiment 2：
The bagasse crushed after drying is placed in glass reactor, sodium hydroxide and the matter that mass fraction is 1% is added
Measure the hydrogen peroxide that score is 5%, solid-to-liquid ratio 1:35(g:G), reaction temperature is 30 DEG C, and the reaction time is for 24 hours, water to be used after filtering
Solid residue is washed to neutrality, obtains pretreatment substrate.With the pretreatment substrate of pH5 buffer solutions dispersion gained, every gram is then pressed
10 family's endo-xylanase of glycoside hydrolase of 400IU is added in butt substrate, and 12h is reacted under the conditions of 50 DEG C and carries out enzyme water
Solution, is obtained by filtration solid residue after xylo-oligosaccharide solution and xylanase hydrolysis.With the zytase of pH5 buffer solutions dispersion gained
Then solid residue after hydrolysis presses every gram of butt substrate and 25IU cellulases is added, 48h is reacted under the conditions of 50 DEG C and carries out enzyme water
Solution, is obtained by filtration fermentable sugars solution.Show that xylo-oligosaccharide conversion ratio is 20.05% by high performance liquid chromatography measurement, grape
Sugared conversion ratio is 82.26%, and xylose rate is 25.31%.
Embodiment 3：
The bagasse crushed after drying is placed in glass reactor, sodium hydroxide and the matter that mass fraction is 1% is added
Measure the hydrogen peroxide that score is 1.5%, solid-to-liquid ratio 1:25(g:G), reaction temperature is 90 DEG C, and reaction time 1h is used after filtering
Water washing solid residue obtains pretreatment substrate to neutrality.With the pretreatment substrate of pH5 buffer solutions dispersion gained, then press every
11 family's endo-xylanase of glycoside hydrolase of 600IU is added in gram butt substrate, and 9h is reacted under the conditions of 55 DEG C and carries out enzyme water
Solution, is obtained by filtration solid residue after xylo-oligosaccharide solution and xylanase hydrolysis.With the zytase of pH5 buffer solutions dispersion gained
Then solid residue after hydrolysis presses every gram of butt substrate and 20IU cellulases is added, 96h is reacted under the conditions of 50 DEG C and carries out enzyme water
Solution, is obtained by filtration fermentable sugars solution.Show that xylo-oligosaccharide conversion ratio is 23.65% by high performance liquid chromatography measurement, grape
Sugared conversion ratio is 85.14%, and xylose rate is 25.16%.
Embodiment 4：
By crush dry after wheat stalk be placed in glass reactor, be added mass fraction be 2% potassium hydroxide and
The hydrogen peroxide that mass fraction is 0.5%, solid-to-liquid ratio 1:25(g:G), reaction temperature is 80 DEG C, reaction time 6h, after filtering
Solid residue is washed with water to neutrality, obtains pretreatment substrate.With the pretreatment substrate of pH5 buffer solutions dispersion gained, then press
11 family's endo-xylanase of glycoside hydrolase of 1000IU is added in every gram of butt substrate, and 6h is reacted under the conditions of 50 DEG C and carries out enzyme
Hydrolysis, is obtained by filtration solid residue after xylo-oligosaccharide solution and xylanase hydrolysis.With the xylan of pH5 buffer solutions dispersion gained
Then solid residue after enzyme hydrolysis presses every gram of butt substrate and 50IU cellulases is added, 96h is reacted under the conditions of 45 DEG C and carries out enzyme
Hydrolysis, is obtained by filtration fermentable sugars solution.Show that xylo-oligosaccharide conversion ratio is 22.35% by high performance liquid chromatography measurement, Portugal
Grape sugar conversion ratio is 82.06%, and xylose rate is 25.17%.
Embodiment 5：
The maize straw crushed after drying is placed in glass reactor, ammonium hydroxide and the quality that mass fraction is 5% is added
The hydrogen peroxide that score is 2%, solid-to-liquid ratio 1:15(g:G), reaction temperature is 80 DEG C, and reaction time 6h is washed with water after filtering
Solid residue is washed to neutrality, obtains pretreatment substrate.With the pretreatment substrate of pH5 buffer solutions dispersion gained, then presses every gram and do
10 family's endo-xylanase of glycoside hydrolase of 100IU is added in substrate, and 12h is reacted under the conditions of 55 DEG C and carries out enzyme hydrolysis,
Solid residue after xylo-oligosaccharide solution and xylanase hydrolysis is obtained by filtration.With the zytase water of pH5 buffer solutions dispersion gained
Then solid residue after solution presses every gram of butt substrate and 50IU/g cellulases is added, 48h is reacted under the conditions of 55 DEG C and carries out enzyme water
Solution, is obtained by filtration fermentable sugars solution.Show that xylo-oligosaccharide conversion ratio is 18.35% by high performance liquid chromatography measurement, grape
Sugared conversion ratio is 72.18%, and xylose rate is 20.03%.
Embodiment 6：
The corncob crushed after drying is placed in glass reactor, sodium hydroxide and the matter that mass fraction is 1% is added
Measure the hydrogen peroxide that score is 1%, solid-to-liquid ratio 1:25(g:G), reaction temperature is 60 DEG C, and reaction time 8h uses water after filtering
Solid residue is washed to neutrality, obtains pretreatment substrate.With the pretreatment substrate of pH5 buffer solutions dispersion gained, every gram is then pressed
10 family's endo-xylanase of glycoside hydrolase of 250IU is added in butt substrate, and 16h is reacted under the conditions of 45 DEG C and carries out enzyme water
Solution, is obtained by filtration solid residue after xylo-oligosaccharide solution and xylanase hydrolysis.With the zytase of pH5 buffer solutions dispersion gained
Then solid residue after hydrolysis presses every gram of butt substrate and 10IU/g cellulases is added, 72h is reacted under the conditions of 50 DEG C and carries out enzyme
Hydrolysis, is obtained by filtration fermentable sugars solution.Show that xylo-oligosaccharide conversion ratio is 20.35% by high performance liquid chromatography measurement, Portugal
Grape sugar conversion ratio is 78.06%, and xylose rate is 22.43%.
Above-listed detailed description is illustrating for possible embodiments of the present invention, which is not to limit this hair
Bright the scope of the claims, all equivalence enforcements or change without departing from carried out by the present invention are intended to be limited solely by the scope of patent protection of this case
In.
Claims (4)
1. a kind of method preparing xylo-oligosaccharide and fermentable sugars using lignocellulose-like biomass, which is characterized in that the party
Method includes the following steps：
(1) by crush dry after lignocellulose-like biomass be placed in glass reactor, be added mass fraction be 0.5%~
The hydrogen peroxide that 5% lye and mass fraction is 0.5%~5%, control solid-liquid mass ratio are 1:10~35, reaction temperature 30
DEG C~90 DEG C, the reaction time is 1h~for 24 hours, is separated by solid-liquid separation after reaction, and solid residue is washed with water to neutrality；
(2) use the solid residue obtained by pH5.0 buffer solutions dispersion steps (1), then press every gram of butt substrate be added 100~
The endo-xylanase of 1000IU, under the conditions of 45~55 DEG C react 6~carry out enzyme hydrolysis for 24 hours, be separated by solid-liquid separation after reaction,
Obtain xylo-oligosaccharide solution and solid residue；
(3) solid residue obtained by pH5.0 buffer solutions dispersion steps (2) is used, every gram of butt substrate is then pressed and 10~50IU is added
Cellulase reacts 48~96h progress enzyme hydrolysis under the conditions of 45~55 DEG C, is separated by solid-liquid separation after reaction, obtains to ferment
Sugar juice.
2. the method according to claim 1 for preparing xylo-oligosaccharide and fermentable sugars using lignocellulose-like biomass,
It is characterized in that, the lignocellulose-like biomass in bagasse, wheat stalk, maize straw or corncob one
Kind.
3. the side according to claim 1 or 2 for preparing xylo-oligosaccharide and fermentable sugars using lignocellulose-like biomass
Method, which is characterized in that the one kind of alkali in sodium hydroxide, potassium hydroxide or ammonium hydroxide in the step (1).
4. the side according to claim 1 or 2 for preparing xylo-oligosaccharide and fermentable sugars using lignocellulose-like biomass
Method, which is characterized in that the one kind of endo-xylanase in glycoside hydrolase Families 10 or 11 in the step (2).
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Cited By (10)
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|---|---|---|---|---|
| CN110016488A (en) * | 2019-04-18 | 2019-07-16 | 中国科学院广州能源研究所 | The method of separating lignin coproduction xylo-oligosaccharide from lignocellulosic alkaline oxygenated pretreatment fluid |
| CN111184225A (en) * | 2020-02-25 | 2020-05-22 | 桂林古膳食品科技有限公司 | High-activity dietary fiber composition and application thereof |
| CN112111541A (en) * | 2019-06-20 | 2020-12-22 | 南京理工大学 | Alkali method for pretreating densified lignocellulose and biotransformation method |
| US11006658B2 (en) | 2018-08-15 | 2021-05-18 | Cambridge Glycoscience Ltd | Compositions, their use, and methods for their formation |
| CN113042096A (en) * | 2021-04-02 | 2021-06-29 | 中国科学院广州能源研究所 | Method for preparing high-quality bio-based polyol by liquefying and hydrogenating lignocellulose by one-pot method |
| US11248247B2 (en) | 2018-02-21 | 2022-02-15 | Cambridge Glycoscience Ltd | Methods and systems of producing oligosaccharides |
| US11297865B2 (en) | 2019-08-16 | 2022-04-12 | Cambridge Glycoscience Ltd | Methods of treating biomass to produce oligosaccharides and related compositions |
| CN114621987A (en) * | 2022-02-22 | 2022-06-14 | 国家粮食和物资储备局科学研究院 | Method for preparing arabinoxylan with different molecular weight distribution characteristics |
| US11871763B2 (en) | 2019-12-12 | 2024-01-16 | Cambridge Glycoscience Ltd | Low sugar multiphase foodstuffs |
| WO2024062148A1 (en) * | 2022-09-19 | 2024-03-28 | Universidade De Vigo | Process for producing hemicellulose-derived products |
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Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11248247B2 (en) | 2018-02-21 | 2022-02-15 | Cambridge Glycoscience Ltd | Methods and systems of producing oligosaccharides |
| US11596165B2 (en) | 2018-08-15 | 2023-03-07 | Cambridge Glycoscience Ltd | Compositions, their use, and methods for their formation |
| US11903399B2 (en) | 2018-08-15 | 2024-02-20 | Cambridge Glycoscience Ltd | Compositions, their use, and methods for their formation |
| US11006658B2 (en) | 2018-08-15 | 2021-05-18 | Cambridge Glycoscience Ltd | Compositions, their use, and methods for their formation |
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| CN112111541A (en) * | 2019-06-20 | 2020-12-22 | 南京理工大学 | Alkali method for pretreating densified lignocellulose and biotransformation method |
| US11297865B2 (en) | 2019-08-16 | 2022-04-12 | Cambridge Glycoscience Ltd | Methods of treating biomass to produce oligosaccharides and related compositions |
| US11771123B2 (en) | 2019-08-16 | 2023-10-03 | Cambridge Glycoscience Ltd | Methods for treating biomass to produce oligosaccharides and related compositions |
| US11871763B2 (en) | 2019-12-12 | 2024-01-16 | Cambridge Glycoscience Ltd | Low sugar multiphase foodstuffs |
| CN111184225A (en) * | 2020-02-25 | 2020-05-22 | 桂林古膳食品科技有限公司 | High-activity dietary fiber composition and application thereof |
| CN113042096B (en) * | 2021-04-02 | 2023-06-09 | 中国科学院广州能源研究所 | Method for preparing high-quality bio-based polyol by lignocellulose one-pot liquefaction and hydrogenation |
| CN113042096A (en) * | 2021-04-02 | 2021-06-29 | 中国科学院广州能源研究所 | Method for preparing high-quality bio-based polyol by liquefying and hydrogenating lignocellulose by one-pot method |
| CN114621987A (en) * | 2022-02-22 | 2022-06-14 | 国家粮食和物资储备局科学研究院 | Method for preparing arabinoxylan with different molecular weight distribution characteristics |
| WO2024062148A1 (en) * | 2022-09-19 | 2024-03-28 | Universidade De Vigo | Process for producing hemicellulose-derived products |
| ES2966032A1 (en) * | 2022-09-19 | 2024-04-17 | Univ Vigo | Production process of products derived from hemicellulose |
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