CN110172487A - The extracting method of withy Ji hemicellulose with arabinose and uronic acid branch - Google Patents

The extracting method of withy Ji hemicellulose with arabinose and uronic acid branch Download PDF

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CN110172487A
CN110172487A CN201910505151.5A CN201910505151A CN110172487A CN 110172487 A CN110172487 A CN 110172487A CN 201910505151 A CN201910505151 A CN 201910505151A CN 110172487 A CN110172487 A CN 110172487A
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hemicellulose
withy
arabinose
uronic acid
extracting method
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丁金花
孟宪智
郁崇文
张斌
杨飞
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Donghua University
National Dong Hwa University
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    • C12P19/00Preparation of compounds containing saccharide radicals
    • C12P19/04Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00Preparation of compounds containing saccharide radicals
    • C12P19/14Preparation 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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Abstract

The present invention provides a kind of extracting methods of withy Ji hemicellulose with arabinose and uronic acid branch, comprising: withy Ji powder is added in peracetic acid soln and carries out obtaining holocellulose after delignification processing;Enzymolysis processing is carried out to holocellulose with cellulase, ethanol solution is added in supernatant, precipitating obtains thick hemicellulose;Thick hemicellulose obtained goes out by dialysis purification obtains the withy Ji hemicellulose with arabinose and uronic acid branch.The method of present invention cellulase hydrolysis releases hemicellulose from holocellulose, and hemicellulose after purification is known as higher molecular weight and the degree of polymerization, remains more functional group and xylan branched structure.For understanding pretreatment to biomass structure, the influence of especially hemicellulose structure has very great help the present invention, is conducive to the research of interaction relationship between the component in biomass cells wall.

Description

The extracting method of withy Ji hemicellulose with arabinose and uronic acid branch
Technical field
The invention belongs to biorefinery fields, are related to a kind of withy Ji half fiber with arabinose and uronic acid branch Tie up the extracting method of element.
Background technique
The separation of the big component of biomass three (cellulose, hemicellulose and lignin) is for grasping biomass structure feature extremely It closes important.Withy Ji is the raw material of most potential second generation bio-fuel and production alternative energy source.Hemicellulose is biology Content is only second to the component of cellulose in matter, in order to preferably be used, understands its physical and chemical performance, it is necessary to by half fiber Dimension element is completely extracted from plant cell wall.Compared to cellulose and lignin, the hemicellulose degree of polymerization is low, and branch is multiple It is miscellaneous, it is often connected with each other with lignin and cellulose etc., therefore can extract what structure was not damaged completely currently without method Hemicellulose.Xylan is the most important a kind of polysaccharide of hemicellulose, is widely used in chemistry, biomaterial and medical accessory etc. Field.The application that characterization is conducive to hemicellulose, functional group are carried out to the complete hemicellulose structure containing branch and functional group Type and quantity directly affect the effective use of biomass, the physical and chemical performance of hemicellulose can be reflected, while also can Influence its degree of reaction and dissolubility.Arabinose and uronic acid in xylan branch can pass through the fiber of hydrogen bond and amorphous region β-Isosorbide-5-Nitrae-glucose chain link on the micro- fibril of element reduces cellulose crystallity, therefore its arabinose and alditol together The degree of substitution of acid is two key factors for improving biomass enzymolysis efficiency.
The structure of hemicellulose depends primarily on species and other environmental factors.The hemicellulose of withy Ji be typically with The xylan that β -1,4 couples is the symmetrical structure of main chain.The extracting method of hemicellulose mainly has alkali soluble solution, You Jirong at present Agent extraction method, Hot water extraction and biochemistry are combined method, these methods have certain limitation.Alkali soluble solution is using most More hemicellulose separation methods, cellulose swollen in aqueous slkali, cell wall are broken, and uronic acid and acetic acid ester bond are by enzyme Solution, so that hemicellulose, lignin and colloid dissolution.But alkali process can destroy ester bond, interrupt and be connected to xylan bone Acetyl group and other branches on frame, reduce the solubility of hemicellulose, then influence its stability.Opposite, dimethyl is sub- The extent of exfoliation of the hemicellulose that the organic solvents such as sulfone, dioxane extract, ester bond is smaller, but its yield (~16%) Well below alkali soluble solution (72%~85%).Cellulase and hemicellulase have specificity to the hydrolysis of polysaccharide, have Research shows that enzymatic hydrolysis and alkali soluble uncoupling usage can separate cellulose and hemicellulose.Pass through zytase/endoglucanase The two-step method that hydrolysis and alkali soluble solution combine can selectively extract high-purity from leaf wood and (account for about hemicellulose total amount 60%), the xylan of high molecular weight.Nevertheless, these extracting methods all inevitably destroy the knot of hemicellulose Structure, yield is low or specificity is insufficient.
Summary of the invention
Technical problem to be solved by the invention is to provide a kind of withy Jis with arabinose and uronic acid branch half The extracting method of cellulose.
In order to solve the above-mentioned technical problems, the present invention provides a kind of withy Ji with arabinose and uronic acid branch The extracting method of hemicellulose, which comprises the following steps:
Step 1: withy Ji raw material being smashed with pulverizer, obtains withy Ji powder;
Step 2: withy Ji powder is continuously used into toluene/ethanol solution and acetone extraction, it is dry, obtain no extract timber Raw material;
Step 3: delignification processing is carried out to no extract wood raw material with Peracetic acid, is washed, it is dry, withy is made Ji holocellulose;
Step 4: withy Ji holocellulose ball grinds are digested with cellulase solution;
Step 5: the supernatant being centrifuged after being digested is added in ethanol water and carries out alcohol precipitation, is then centrifuged for being sunk Forming sediment is thick hemicellulose enzymolysis;
Step 6: by thick hemicellulose enzymolysis by dialysing, freezing, be dried to obtain with arabinose and uronic acid branch The withy Ji hemicellulose of chain.
Preferably, the smashed withy Ji powder in the step 1 crosses a 0.5mm sieve.
Preferably, method for extracting is soxhlet type in the step 2.
It is highly preferred that extraction conditions in the step 2 are as follows: 85 DEG C of temperature, extract 8h with toluene/ethanol mixed solution Afterwards, then with acetone extraction 4h.
Further, in the toluene/ethanol mixed solution, VToluene: VEthyl alcohol=2:1.
Preferably, in the step 3 delignification treatment conditions are as follows: peroxide will be added to without extract wood raw material In acetic acid aqueous solution, treatment temperature is 25 DEG C, and the time is for 24 hours;Mass ratio wherein without extract wood raw material and Peracetic acid is 1:(3.5~5.5), the quality of no extract wood raw material accounts for the 10~15% of reaction system gross mass.
It is highly preferred that the operation of the step 2, step 3 carries out under ventilation condition.
Preferably, it is washed in the step 3 are as follows: filtered, and be washed with deionized water with 0.45 μm of water phase micropore filtering film Washing to pH value is 7.0.
Preferably, ball milling condition in the step 4 are as follows: select the ball grinder and ball-type zirconium oxide abrasive of cobalt oxide material The revolving speed of ball, Ball-milling Time 3h, ball mill is set as 600rpm, and it is ball milling 5min that a cyclic control system, which is arranged, cooling 5min。
Preferably, the condition digested in the step 4 are as follows: by the withy Ji holocellulose powder after ball milling be added to containing The sodium acetate buffer of cellulase carries out enzymolysis processing, and the specification of sodium acetate buffer is 50mM, pH 5.0, herein Reason carries out in shaking table, and temperature is 37 DEG C, revolving speed 200rpm, and the processing time is 72h.
It is highly preferred that the mass ratio of the withy Ji holocellulose powder and cellulase is 1000:(5.4~30).
It is highly preferred that the cellulase selected is buying in Sigma-Aldrich, the Trichoderma reesei of Inc. ATCC 26921。
Preferably, in the step 5 alcohol precipitation condition are as follows: supernatant is slowly dropped into 65% (v:v) second of 4 times of volumes In alcohol solution, the lower ethanol water of concentration, which can screen, is settled out the biggish hemicellulose of molecular weight, removes glucose etc. Small-molecule substance, the cool overnight in 4 DEG C of refrigerators, is collected by centrifugation precipitating.
It is highly preferred that the condition being centrifuged in the step 5: revolving speed is set as 8000rpm, time 15min.
Preferably, the step 6 specifically: thick hemicellulose enzymolysis is added in deionized water, is transferred to bag filter In, at least 72h is placed in the deionized water of flowing, and then dialyzate after purification is placed in -20 DEG C of refrigerators or in liquid nitrogen Freezing is put into freeze dryer dry 48h, the withy Ji hemicellulose with arabinose and uronic acid branch is made.
It is highly preferred that the dialysis selection is purchased in the 3.5K MWCO of Thermo Fisher scientific Thermo Scientific dialysis machine.
Compared with prior art, the beneficial effects of the present invention are:
1, the method for the novel cellulase degradation of one kind of the invention, in conjunction with the processing skill of Peracetic acid removal lignin Art extracts the small withy Ji hemicellulose of structural damage, is conducive to the structure feature for grasping natural hemicellulose, especially I The branch chain information of uncle sugar and uronic acid, so as to more effectively be used.
2, hemicellulose extracting method provided by the invention can retain the function in hemicellulose macromolecular to the maximum extent Group facilitates the interaction relationship for understanding hemicellulose and other cell-wall components.
Specific embodiment
Present invention will be further explained below with reference to specific examples.It should be understood that these embodiments are merely to illustrate the present invention Rather than it limits the scope of the invention.In addition, it should also be understood that, after reading the content taught by the present invention, those skilled in the art Member can make various changes or modifications the present invention, and such equivalent forms equally fall within the application the appended claims and limited Range.
The present invention is suitable for any withy Ji kind, and the withy Ji that various embodiments of the present invention are selected is by Samuel Roberts Nobel Foundation is provided, and plantation in 2011 was harvested in the area Ardmore, 2012;The cellulase of selection is buying In Sigma-Aldrich, the Trichoderma reesei ATCC 26921 of Inc.;What dialysis was selected is buying in Thermo The Thermo Scientific dialysis machine of the 3.5K MWCO of Fisher scientific.
Embodiment 1
Present embodiments provide a kind of withy Ji hemicellulose extracting method of cellulase auxiliary, specific steps are as follows:
(1) withy Ji raw material is crushed, crosses 0.5mm sieve, obtains withy Ji powder;
(2) in draught cupboard, with soxhlet extraction methods (condition are as follows: 85 DEG C of temperature, taken out with the toluene/ethanol that volume ratio is 2:1 After mentioning 8h, then with the extractables such as lipid, wax in acetone extraction 4h) removal raw material, take out the powder extracted be placed in it is logical It is dry under room temperature in wind cupboard, it obtains without extract solid material;
(3) in draught cupboard, delignification processing is carried out without extracting raw material to dry: weighing 328g Peracetic acid The aqueous solution that deionized water is configured to Peracetic acid to 360g is added in (concentration 32wt%), and being placed in volume is 1000mL tool plug In conical flask, it is put into magnet rotor, the 360g peroxide acetate aqueous solution prepared is added without extracting raw material after dry in 30g In, it is reacted for 24 hours, is filtered by 0.45 μm of water phase micropore filtering film, with a large amount of deionized waters at room temperature using magnetic stirring apparatus Washing to the pH value of solution is 7.0 or so, obtains withy Ji holocellulose;
(4) the withy Ji holocellulose is filtered out, air-dries, is clayed into power with PA-100 planetary type ball-milling instrument, the time The revolving speed of 3h, ball mill are set as 600rpm, and the circulation that control system is arranged is ball milling 5min, and cooling 5min takes 3g ball Holocellulose powder after mill is added to the sodium acetate buffer that 40mL contains 90mg cellulase and carries out enzymolysis processing, acetic acid The specification of sodium buffer solution is 50mM, pH 5.0, this processing carries out in shaking table, and temperature is 37 DEG C, revolving speed 200rpm, processing Time is 72h;
(5) supernatant of the enzymatic hydrolysis, centrifugal condition are obtained by centrifuging and taking are as follows: revolving speed is set as 8000rpm, time 15min;The supernatant of above-mentioned enzymatic hydrolysis, is added to the water of 65% ethyl alcohol of 4 times of volumes by the ethanol water for preparing 65% (v:v) In solution, it is statically placed in cool overnight in 4 DEG C of refrigerators, centrifugation obtains and is precipitated as thick hemicellulose enzymolysis;
(6) 5mL deionized water is added in thick hemicellulose enzymolysis, is transferred in the bag filter of 3.5K, bag filter is put It sets in 2000mL beaker, at least 72h is placed under the deionized water of flowing, the small molecules such as ethyl alcohol, glucose are constantly set It swaps out and, dialyzate is then placed in -20 DEG C of refrigerators (or in liquid nitrogen) freezing, be put into freeze dryer at least dry 48h, system There must be the withy Ji hemicellulose of arabinose and uronic acid branch.
Its yield is tested with weight method, as a result sees attached list 1.
With its monosaccharide of ion chromatography and glucuronic acid content, its molecular weight and molecular weight point are tested with gel permeation chromatography As a result cloth sees attached list 2~3.
Embodiment 2
Present embodiments provide a kind of withy Ji hemicellulose extracting method of cellulase auxiliary, specific steps are as follows:
(1) withy Ji raw material is crushed, crosses 0.5mm sieve, obtains withy Ji powder;
(2) in draught cupboard, with soxhlet extraction methods (condition are as follows: 85 DEG C of temperature, taken out with the toluene/ethanol that volume ratio is 2:1 After mentioning 8h, then with the extractables such as lipid, wax in acetone extraction 4h) removal raw material, take out the powder extracted be placed in it is logical It is dry under room temperature in wind cupboard, it obtains without extract solid material;
(3) in draught cupboard, delignification processing is carried out without extracting raw material to dry: weighing 328g Peracetic acid The aqueous solution that deionized water is configured to Peracetic acid to 360g is added in (concentration 32wt%), and being placed in volume is 1000mL tool plug In conical flask, it is put into magnet rotor, the 360g peroxide acetate aqueous solution prepared is added without extracting raw material after dry in 30g In, it is reacted for 24 hours, is filtered by 0.45 μm of water phase micropore filtering film, with a large amount of deionized waters at room temperature using magnetic stirring apparatus Washing to the pH value of solution is 7.0 or so, obtains withy Ji holocellulose;
(4) the withy Ji holocellulose is filtered out, air-dries, takes 3g holocellulose powder to be added to 40mL and contain 90mg The sodium acetate buffer of cellulase carries out enzymolysis processing, and the specification of sodium acetate buffer is 50mM, pH 5.0, herein Reason carries out in shaking table, and temperature is 37 DEG C, revolving speed 200rpm, and the processing time is 72h;
(5) supernatant of the enzymatic hydrolysis, centrifugal condition are obtained by centrifuging and taking are as follows: revolving speed is set as 8000rpm, time 15min;The supernatant of above-mentioned enzymatic hydrolysis, is added to the water of 65% ethyl alcohol of 4 times of volumes by the ethanol water for preparing 65% (v:v) In solution, it is statically placed in cool overnight in 4 DEG C of refrigerators, centrifugation obtains and is precipitated as thick hemicellulose enzymolysis;
(6) 5mL deionized water is added in thick hemicellulose enzymolysis, is transferred in the bag filter of 3.5K, bag filter is put It sets in 2000mL beaker, at least 72h is placed under the deionized water of flowing, the small molecules such as ethyl alcohol, glucose are constantly set It swaps out and, dialyzate is then placed in -20 DEG C of refrigerators (or in liquid nitrogen) freezing, be put into freeze dryer at least dry 48h, system There must be the withy Ji hemicellulose of arabinose and uronic acid branch.
Its yield is tested with weight method, as a result sees attached list 1.
Embodiment 3
Present embodiments provide a kind of withy Ji hemicellulose extracting method of cellulase auxiliary, specific steps are as follows:
(1) withy Ji raw material is crushed, crosses 0.5mm sieve, obtains withy Ji powder;
(2) in draught cupboard, with soxhlet extraction methods (condition are as follows: 85 DEG C of temperature, taken out with the toluene/ethanol that volume ratio is 2:1 After mentioning 8h, then with the extractables such as lipid, wax in acetone extraction 4h) removal raw material, take out the powder extracted be placed in it is logical It is dry under room temperature in wind cupboard, it obtains without extract solid material;
(3) in draught cupboard, delignification processing is carried out without extracting raw material to dry: weighing 328g Peracetic acid The aqueous solution that deionized water is configured to Peracetic acid to 360g is added in (concentration 32wt%), and being placed in volume is 1000mL tool plug In conical flask, it is put into magnet rotor, the 360g peroxide acetate aqueous solution prepared is added without extracting raw material after dry in 30g In, it is reacted for 24 hours, is filtered by 0.45 μm of water phase micropore filtering film, with a large amount of deionized waters at room temperature using magnetic stirring apparatus Washing to the pH value of solution is 7.0 or so, obtains withy Ji holocellulose;
(4) the withy Ji holocellulose is filtered out, air-dries, takes 3g holocellulose powder to be added to 40mL and contain 54mg The sodium acetate buffer of cellulase carries out enzymolysis processing, and the specification of sodium acetate buffer is 50mM, pH 5.0, herein Reason carries out in shaking table, and temperature is 37 DEG C, revolving speed 200rpm, and the processing time is 72h;
(5) supernatant of the enzymatic hydrolysis, centrifugal condition are obtained by centrifuging and taking are as follows: revolving speed is set as 8000rpm, time 15min;The supernatant of above-mentioned enzymatic hydrolysis, is added to the water of 65% ethyl alcohol of 4 times of volumes by the ethanol water for preparing 65% (v:v) In solution, it is statically placed in cool overnight in 4 DEG C of refrigerators, centrifugation obtains and is precipitated as thick hemicellulose enzymolysis;
(6) 5mL deionized water is added in thick hemicellulose enzymolysis, is transferred in the bag filter of 3.5K, bag filter is put It sets in 2000mL beaker, at least 72h is placed under the deionized water of flowing, the small molecules such as ethyl alcohol, glucose are constantly set It swaps out and, dialyzate is then placed in -20 DEG C of refrigerators (or in liquid nitrogen) freezing, be put into freeze dryer at least dry 48h, system There must be the withy Ji hemicellulose of arabinose and uronic acid branch.
Its yield is tested with weight method, as a result sees attached list 1.
Embodiment 4
Present embodiments provide a kind of withy Ji hemicellulose extracting method of cellulase auxiliary, specific steps are as follows:
(1) withy Ji raw material is crushed, crosses 0.5mm sieve, obtains withy Ji powder;
(2) in draught cupboard, with soxhlet extraction methods (condition are as follows: 85 DEG C of temperature, taken out with the toluene/ethanol that volume ratio is 2:1 After mentioning 8h, then with the extractables such as lipid, wax in acetone extraction 4h) removal raw material, take out the powder extracted be placed in it is logical It is dry under room temperature in wind cupboard, it obtains without extract solid material;
(3) in draught cupboard, delignification processing is carried out without extracting raw material to dry: weighing 328g Peracetic acid The aqueous solution that deionized water is configured to Peracetic acid to 360g is added in (concentration 32wt%), and being placed in volume is 1000mL tool plug In conical flask, it is put into magnet rotor, the 360g peroxide acetate aqueous solution prepared is added without extracting raw material after dry in 30g In, it is reacted for 24 hours, is filtered by 0.45 μm of water phase micropore filtering film, with a large amount of deionized waters at room temperature using magnetic stirring apparatus Washing to the pH value of solution is 7.0 or so, obtains withy Ji holocellulose;
(4) the withy Ji holocellulose is filtered out, air-dries, takes 3g holocellulose powder to be added to 40mL and contain The sodium acetate buffer of 16.2mg cellulase carries out enzymolysis processing, and the specification of sodium acetate buffer is 50mM, and pH is 5.0, this processing carries out in shaking table, and temperature is 37 DEG C, revolving speed 200rpm, and the processing time is 72h;
(5) supernatant of the enzymatic hydrolysis, centrifugal condition are obtained by centrifuging and taking are as follows: revolving speed is set as 8000rpm, time 15min;The supernatant of above-mentioned enzymatic hydrolysis, is added to the water of 65% ethyl alcohol of 4 times of volumes by the ethanol water for preparing 65% (v:v) In solution, it is statically placed in cool overnight in 4 DEG C of refrigerators, centrifugation obtains and is precipitated as thick hemicellulose enzymolysis;
(6) 5mL deionized water is added in thick hemicellulose enzymolysis, is transferred in the bag filter of 3.5K, bag filter is put It sets in 2000mL beaker, at least 72h is placed under the deionized water of flowing, the small molecules such as ethyl alcohol, glucose are constantly set It swaps out and, dialyzate is then placed in -20 DEG C of refrigerators (or in liquid nitrogen) freezing, be put into freeze dryer at least dry 48h, system There must be the withy Ji hemicellulose of arabinose and uronic acid branch.
Its yield is tested with weight method, as a result sees attached list 1.
Comparative example 1
This comparative example provides a kind of method for extracting withy Ji hemicellulose by organic solvent dimethyl sulfoxide, specifically Step are as follows:
(1) withy Ji raw material is crushed, crosses 0.5mm sieve, obtains withy Ji powder;
(2) in draught cupboard, with soxhlet extraction methods (condition are as follows: 85 DEG C of temperature, taken out with the toluene/ethanol that volume ratio is 2:1 After mentioning 8h, then with the extractables such as lipid, wax in acetone extraction 4h) removal raw material, take out the powder extracted be placed in it is logical It is dry under room temperature in wind cupboard, it obtains without extract solid material;
(3) in draught cupboard, delignification processing is carried out without extracting raw material to dry: weighing 328g Peracetic acid The aqueous solution that deionized water is configured to Peracetic acid to 360g is added in (concentration 32wt%), and being placed in volume is 1000mL tool plug In conical flask, it is put into magnet rotor, the 360g peroxide acetate aqueous solution prepared is added without extracting raw material after dry in 30g In, it is reacted for 24 hours, is filtered by 0.45 μm of water phase micropore filtering film, with a large amount of deionized waters at room temperature using magnetic stirring apparatus Washing to the pH value of solution is 7.0 or so, obtains withy Ji holocellulose;
(4) the withy Ji holocellulose is filtered out, is air-dried, take 3g holocellulose powder that the dimethyl of 120mL is added Sulfoxide (DMSO), nitrogen charging gas shielded come into full contact with holocellulose with DMSO with magnetic agitation, and extraction temperature is 70 DEG C, the time For 5h;
(5) alkali lysate is obtained by filtration with 0.45 μm of oily phase filter membrane, 20mL deionized water is added to wash solid residue, it will PH value is adjusted to 3.5 using glacial acetic acid after DMSO lysate and washing lotion mixing, and the DMSO lysate among the above with after is added to 4 In 95% ethanol solution of times volume, it is statically placed in cool overnight in 4 DEG C of refrigerators, centrifugation obtains and is precipitated as thick DMSO hemicellulose Element, centrifugal condition: revolving speed is set as 8000rpm, time 15min;
(6) 5mL deionized water is added in thick DMSO hemicellulose, is transferred in the bag filter of 3.5K, bag filter is put It sets in 2000mL beaker, at least 72h is placed under the deionized water of flowing, the small molecules such as ethyl alcohol, glacial acetic acid, DMSO are continuous Ground cements out, and dialyzate is then placed in -20 DEG C of refrigerators (or in liquid nitrogen) freezing, is put into freeze dryer at least dry Pure DMSO withy Ji hemicellulose is made in 48h.
With its monosaccharide of ion chromatography and glucuronic acid content, its molecular weight and molecular weight point are tested with gel permeation chromatography As a result cloth sees attached list 2~3.
Comparative example 2
This comparative example provides a kind of method for extracting withy Ji hemicellulose by traditional alkali soluble solution, specific steps Are as follows:
(1) withy Ji raw material is crushed, crosses 0.5mm sieve, obtains withy Ji powder;
(2) in draught cupboard, with soxhlet extraction methods (condition are as follows: 85 DEG C of temperature, taken out with the toluene/ethanol that volume ratio is 2:1 After mentioning 8h, then with the extractables such as lipid, wax in acetone extraction 4h) removal raw material, take out the powder extracted be placed in it is logical It is dry under room temperature in wind cupboard, it obtains without extract solid material;
(3) in draught cupboard, delignification processing is carried out without extracting raw material to dry: weighing 328g Peracetic acid The aqueous solution that deionized water is configured to Peracetic acid to 360g is added in (concentration 32wt%), and being placed in volume is 1000mL tool plug In conical flask, it is put into magnet rotor, the 360g peroxide acetate aqueous solution prepared is added without extracting raw material after dry in 30g In, it is reacted for 24 hours, is filtered by 0.45 μm of water phase micropore filtering film, with a large amount of deionized waters at room temperature using magnetic stirring apparatus Washing to the pH value of solution is 7.0 or so, obtains withy Ji holocellulose;
(4) the withy Ji holocellulose is filtered out, air-dry, take 3g holocellulose powder be added 36g mass fraction be 17.5% sodium hydroxide (NaOH) solution, nitrogen charging gas shielded come into full contact with holocellulose with lye with magnetic agitation, extracting Time is 16h;
(5) alkali lysate is obtained by filtration with 0.45 μm of oily phase filter membrane, 20mL deionized water is added to wash solid residue, it will PH value is adjusted to 5.5 using glacial acetic acid after DMSO lysate and washing lotion mixing, and the alkali lysate among the above with after is added to 4 times In 95% ethanol solution of volume, it is statically placed in cool overnight in 4 DEG C of refrigerators, centrifugation obtains and is precipitated as thick alkali hemicellulose, from Heart condition: revolving speed is set as 8000rpm, time 15min;
(6) 5mL deionized water is added in thick alkali hemicellulose, is transferred in the bag filter of 3.5K, bag filter is placed In 2000mL beaker, at least 72h is placed under the deionized water of flowing, the small molecules such as ethyl alcohol, glacial acetic acid are constantly replaced Out, dialyzate is then placed in -20 DEG C of refrigerators (or in liquid nitrogen) freezing, is put into freeze dryer at least dry 48h, is made Pure alkali withy Ji hemicellulose.
With its monosaccharide of ion chromatography and glucuronic acid content, its molecular weight and molecular weight point are tested with gel permeation chromatography As a result cloth sees attached list 2~3.
The comparison of 1 hemicellulose chemical component test result of subordinate list
* yield is calculated based on the content without hemicellulose in extracting raw material.
The test result of 2 hemicellulose yield of subordinate list, molecular weight and molecular weight distribution compares
* yield is calculated based on the content without hemicellulose in extracting raw material.
The comparison of 3 hemicellulose chemical component test result of subordinate list
* Glc: glucose, Xyl: xylose, Ara: arabinose, Gal: galactolipin, GalA: galacturonic acid, GlaA: Portugal Grape uronic acid
Contain more arabinose and alditol with the withy Ji hemicellulose that the method that cellulase assists is extracted Acid, both usual ingredients are to be connected on hemicellulose xylan skeleton as branch, therefore hemicellulose enzymolysis retains More branches and active group.
The method of cellulase auxiliary of the present invention can be extracted to obtain the hemicellulose of high molecular weight, and polydispersity coefficient Small, molecular weight homogeneity is higher.

Claims (10)

1. a kind of extracting method of the withy Ji hemicellulose with arabinose and uronic acid branch, which is characterized in that including Following steps:
Step 1: withy Ji raw material being smashed with pulverizer, obtains withy Ji powder;
Step 2: withy Ji powder is continuously used into toluene/ethanol solution and acetone extraction, it is dry, it is former to obtain no extract timber Material;
Step 3: delignification processing is carried out to no extract wood raw material with Peracetic acid, is washed, it is dry, it is comprehensive that withy Ji is made Cellulose;
Step 4: withy Ji holocellulose ball grinds are digested with cellulase;
Step 5: the supernatant being centrifuged after being digested is added in ethanol water and carries out alcohol precipitation, is then centrifuged for being precipitated as Thick hemicellulose enzymolysis;
Step 6: by thick hemicellulose enzymolysis by dialysing, freezing, be dried to obtain with arabinose and uronic acid branch Withy Ji hemicellulose.
2. the extracting method of the withy Ji hemicellulose as described in claim 1 with arabinose and uronic acid branch, special Sign is that method for extracting is soxhlet type, extraction conditions in the step 2 are as follows: 85 DEG C of temperature, with toluene/ethanol mixed solution After extracting 8h, then with acetone extraction 4h.
3. the extracting method of the withy Ji hemicellulose as claimed in claim 2 with arabinose and uronic acid branch, special Sign is, in the toluene/ethanol mixed solution, VToluene: VEthyl alcohol=2:1.
4. the extracting method of the withy Ji hemicellulose as described in claim 1 with arabinose and uronic acid branch, special Sign is, the treatment conditions of delignification in the step 3 are as follows: it is water-soluble will to be added to Peracetic acid without extract wood raw material In liquid, treatment temperature is 25 DEG C, and the time is for 24 hours;Mass ratio wherein without extract wood raw material and Peracetic acid be 1:(3.5~ 5.5), the quality of no extract wood raw material accounts for the 10~15% of reaction system gross mass.
5. the extracting method of the withy Ji hemicellulose as described in claim 1 with arabinose and uronic acid branch, special Sign is, washs in the step 3 are as follows: is filtered, and is washed with deionized to pH value with 0.45 μm of water phase micropore filtering film It is 7.0.
6. the extracting method of the withy Ji hemicellulose as described in claim 1 with arabinose and uronic acid branch, special Sign is, ball milling condition in the step 4 are as follows: selects the ball grinder and ball-type zirconium oxide abrasive ball of cobalt oxide material, ball milling The revolving speed of time 3h, ball mill are set as 600rpm, and it is ball milling 5min that a cyclic control system, which is arranged, cooling 5min.
7. the extracting method of the withy Ji hemicellulose as described in claim 1 with arabinose and uronic acid branch, special Sign is, the condition digested in the step 4 are as follows: the withy Ji holocellulose powder after ball milling is added to containing cellulase Sodium acetate buffer carry out enzymolysis processing, the specification of sodium acetate buffer is 50mM, pH 5.0, this processing is in shaking table Middle progress, temperature are 37 DEG C, revolving speed 200rpm, and the processing time is 72h.
8. the extracting method of the withy Ji hemicellulose as claimed in claim 7 with arabinose and uronic acid branch, special Sign is that the mass ratio of the withy Ji holocellulose powder and cellulase is 1000:(5.4~30).
9. the extracting method of the withy Ji hemicellulose as described in claim 1 with arabinose and uronic acid branch, special Sign is, the condition of alcohol precipitation in the step 5 are as follows: supernatant is slowly dropped into 65% (v:v) ethanol water of 4 times of volumes In, precipitating is collected by centrifugation in the cool overnight in 4 DEG C of refrigerators.
10. the extracting method of the withy Ji hemicellulose as described in claim 1 with arabinose and uronic acid branch, special Sign is, the step 6 specifically: thick hemicellulose enzymolysis is added in deionized water, is transferred in bag filter, is flowing Deionized water in place at least 72h, then dialyzate after purification is placed in -20 DEG C of refrigerators or is freezed in liquid nitrogen, is put into Dry 48h, is made the withy Ji hemicellulose with arabinose and uronic acid branch in freeze dryer.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113999331A (en) * 2021-11-17 2022-02-01 华南农业大学 Method for increasing branching degree of hemicellulose by aid of arabinose ball milling and application of method

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102203343A (en) * 2008-09-09 2011-09-28 特里弗里生物量解决方案股份有限公司 Arundo donax hemicellulose recovery process
CN105860090A (en) * 2016-04-25 2016-08-17 北京林业大学 Method for extracting high-activity lignin from biomass and lignin extracted by same
CN106065080A (en) * 2016-04-06 2016-11-02 南京林业大学 A kind of dissolving recycled wood quality extracting method
CN108586771A (en) * 2018-03-30 2018-09-28 中国林业科学研究院 A method of efficiently separating lignin in wood cell wall

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102203343A (en) * 2008-09-09 2011-09-28 特里弗里生物量解决方案股份有限公司 Arundo donax hemicellulose recovery process
CN106065080A (en) * 2016-04-06 2016-11-02 南京林业大学 A kind of dissolving recycled wood quality extracting method
CN105860090A (en) * 2016-04-25 2016-08-17 北京林业大学 Method for extracting high-activity lignin from biomass and lignin extracted by same
CN108586771A (en) * 2018-03-30 2018-09-28 中国林业科学研究院 A method of efficiently separating lignin in wood cell wall

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
BHAGIA,S.等: "Ultrastructure and Enzymatic Hydrolysis of Deuterated Switchgrass", 《ULTRASTRUCTURE AND ENZYMATIC HYDROLYSIS OF DEUTERATED SWITCHGRASS》 *
GENG, W.等: "Effect of delignification on hemicellulose extraction from switchgrass, poplar, and pine and its effect on enzymatic convertibility of cellulose-rich residues", 《BIORES.》 *
SHI,J.等: "Application of cellulase and hemicellulase to pure xylan, pure cellulose, and switchgrass solids from leading pretreatments", 《BIORESOUR.TECHNOL.》 *
段仰凯等: "改进的柳枝稷预处理方法及乙醇发酵研究", 《太阳能学报》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113999331A (en) * 2021-11-17 2022-02-01 华南农业大学 Method for increasing branching degree of hemicellulose by aid of arabinose ball milling and application of method

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