CN105695536A - Pretreatment method of capable of implementing high-efficiency enzymolysis and saccharification on deciduous wood crotches - Google Patents
Pretreatment method of capable of implementing high-efficiency enzymolysis and saccharification on deciduous wood crotches Download PDFInfo
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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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/02—Monosaccharides
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- 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 relates to a pretreatment method of capable of implementing high-efficiency enzymolysis and saccharification on deciduous wood crotches. The method comprises the following steps: cutting deciduous wood crotches into wood chips to obtain a wood chip raw material; putting the deciduous wood crotch wood chip raw material into a boiler to obtain treated wood chips; and carrying out chemical preimpregnation treatment, carrying out two-stage grinding treatment on the treated wood chips, and carrying out bioenzyme constant-temperature culture enzymolysis treatment under aseptic conditions to obtain glucose and xylose, thereby implementing high-efficiency enzyme hydrolysis conversion on the deciduous wood crotches. The method sufficiently implements effective utilization on the forestal waste deciduous wood crotches, thereby effectively utilizing the forestal waste resources, and reducing the pressure on the environmental protection; and thus, the method is an imminent method which is beneficial to both the nation and people and conforms to the national sustainable development strategy, and has important meanings for national economy development, clean production and environmental protection.
Description
Technical field
The invention belongs to biomass resource and utilize technical field with forestry waste, especially a kind of preprocess method realizing the efficient enzyme hydrolysis conversion of leaf wood brush wood。
Background technology
Along with continuing to increase of world population quantity, economical develops rapidly, and non-renewable energy resources (such as oil and natural gas) are constantly consumed, and relevant research worker predicts that the crude oil production in the year two thousand fifty whole world will reduce to 5,000,000,000 barrels from present 25,000,000,000 barrels。Due to bio-fuel in a lot of fields can some or all of substitute fossil fuels, simultaneously national governments also encourage the development of bio-fuel, and this makes people more favor in the exploitation of this regenerative resource and application。
Wood fiber biomass is mainly used in pulping and paper-making industry in traditional industry is applied; but the shortage of global resources and the energy brings crisis not only to other industry; also bring huge challenge to the sector, but this also provides the opportunity of development for pulping and paper-making industry。In " paper industry technological progress " 12 " instruction " that papermaking association of China delivers accordingly, resource and the utilization of biomass is mentioned in many places, and explicitly point out: " paper industry is the renewable material of employing is the processing industry that raw material is largest, in material circulation utilization and low-carbon (LC) production technology and exploitation, there is the advantage of uniqueness ", and emphasize: " industry competition increasingly sharpened makes tradition paper industry face huge pressure, tradition paper mill is converted into and can produce paper pulp and paper simultaneously, macromolecular material, the complex biological matter refinery of chemicals and biomass energy, reach the cellulose in fully reasonable utilizing vegetable fibers at high value raw material, hemicellulose and the big component of lignin three are the new development trend of slurrying scientific research "。It is known that pulp and paper industry is huge to the consumption of biomass resource, therefore, it is trend of the times that biorefinery combines with pulp and paper industry。In biorefinery process, bio-ethanol is a kind of reproducible, green gasoline succedaneum, also will be second biomass consumer goods after paper pulp in the world。Owing to first generation ethanol is to be formed by crops fermentation, the price of grain will be had adverse effect on by this。Therefore, as development of raw materials using cellulose (such as bagasse, straw and timber etc.) and produce second filial generation ethanol and have very important significance。
The trunk of leaf wood is mainly used in timber processing and Furniture manufacture, but can produce substantial amounts of brush wood garbage in the course of processing of timber。If these garbages reasonably not being utilized, not only causing the waste of resource, and polluting the environment。These discarded brush woods are a kind of abundant lignocellulose biomass resources; by these biomass resources of trans-utilization and produce further and a series of there is high value-added product; such as products such as ethanol, carbon fiber, polymer, kerosene and biodiesel, to resource make full use of and environmental conservation is significant。
At present, still do not have a kind of to be capable of in leaf wood brush wood carbohydrate components Efficient Conversion and the method separated。
By retrieving, not yet find the patent publication us relevant to present patent application。
Summary of the invention
It is an object of the invention to overcome the deficiencies in the prior art part, a kind of stubbornly resisting property destroying biomass is provided and the preprocess method that the efficient enzyme hydrolysis of leaf wood brush wood converts can be realized, the method is capable of the efficient utilization of this forestry waste of leaf wood brush wood, and has high value added product offer raw material basis for follow-up processing and utilization or production。
The technical solution adopted for the present invention to solve the technical problems is:
A kind of preprocess method realizing the efficient enzyme hydrolysis conversion of leaf wood brush wood, step is as follows:
(1) first, leaf wood brush wood is cut into wood chip, obtains chip raw material；
(2) leaf wood brush wood chip raw material is placed in boiling vessel, adds the H relative to chip raw material absolute dry mass 0.5% 2.0%2SO4Solid-to-liquid ratio is 1:4 1:10 (kg:L), boiling temperature is incubated 20 90min at the temperature of 120 180 DEG C, and in course of reaction, make the continuous circulated sprinkling of reactant liquor, so that leaf wood brush wood chip raw material and medicinal liquid fully react, after reaction terminates, pretreatment fluid and solid residue are easily separated, reacted wood chip solid residue is washed with water to neutrality, stores for future use after air-dry, obtain wood chip after process；
(3) wood chip after the process after dilute sulfuric acid processes is carried out grinding process, disappear dive process, discongest process and processed after, obtain dehydration disposed slurry, then by dehydration disposed slurry collect and store for follow-up cellulase treatment in 4 DEG C of environment；
(4) cellulase treatment: dehydration disposed slurry is carried out cellulase treatment, before being processed, carries out high temperature sterilize process to container used；
Aseptically, ethyl sodium buffer solution is added in dehydration disposed slurry, the ratio (kg:L) of the absolute dry mass of dehydration disposed slurry and ethyl sodium buffer solution is 1:4 1:6, enzyme mixed liquor is added after being sufficiently stirred for, being subsequently adding water makes the mass concentration of leaf wood slurry be 1.0% 5.0%, carry out biological enzyme hydrolysis process under aseptic condition, monosaccharide component, it is achieved the efficient enzyme hydrolysis of leaf wood brush wood converts；
Wherein, the condition that described biological enzyme hydrolysis processes is: pH4.8, reaction temperature 40 55 DEG C, response time 10 90h, rotating speed 100 180rpm, cellulase consumption used is 10 30FPU/g substrates, and hemicellulose enzyme dosage is 20 80U/g substrates。
And, described step concrete operation step (3) is as follows:
1. wood chip after the process after dilute sulfuric acid processes is placed in airtight reactor, is sequentially added into distilled water, the NaOH relative to chip raw material absolute dry mass 1.0% 5.0% and the Na relative to chip raw material absolute dry mass 1.0% 5.0%2SO3, finally it being made into the mass percentage concentration relative to chip raw material absolute dry mass 20% 50%, this closed reactor is placed in water bath with thermostatic control and carries out Chemical impregnation process, treatment temperature is 60 90 DEG C, processes time 20 40min；
2. after Chemical impregnation processes, after process, wood chip processes then through two sections of grindings: first paragraph is pressure grinding, and actual conditions is: temperature 120 180 DEG C, pressure 0.1 0.5MPa, starches dense 10% 40% (mass percents), grinding 2 10min；Second segment is normal pressure grinding, and actual conditions is: disk mill gap 0.08 0.10mm, starches dense 10% 30% (mass percents), obtains leaf wood slurry；
3. after two sections of grindings process, leaf wood slurry is made into dense 1.0% 5.0% (mass percents) of slurry, under 80 90 DEG C of conditions, it is incubated 10 30min carries out the process of diving that disappears, then the leaf wood slurry after diving that disappears is discongested 10,000 30000 turns, obtain leaf wood pulp fiber；Leaf wood pulp fiber is through screening the fraction being collected by 0.15mm screen slot, it is 20%-40% by the leaf wood slurry recycling dehydration collected to mass percentage concentration, obtain dehydration disposed slurry, then dehydration disposed slurry collected and store for follow-up cellulase treatment in 4 DEG C of environment；
Or, described step concrete operation step (3) is as follows:
1. being placed in grinding device by wood chip after the process after dilute sulfuric acid processes and carry out grinding process, actual conditions is: disk mill gap 0.08 0.10mm, starches dense 10% 30% (mass percents), obtains leaf wood slurry；
2. after grinding processes, leaf wood slurry is made into dense 1.0% 5.0% (mass percents) of slurry, under 80 90 DEG C of conditions, it is incubated 10 30min carries out the process of diving that disappears, then the leaf wood slurry after diving that disappears is discongested 10,000 30000 turns, obtain leaf wood pulp fiber；Leaf wood pulp fiber is through screening the fraction being collected by 0.15mm screen slot, it is 20%-40% by the leaf wood slurry recycling dehydration collected to mass percentage concentration, obtain dehydration disposed slurry, then dehydration disposed slurry collected and store for follow-up cellulase treatment in 4 DEG C of environment；
And, described leaf wood is one or more the mixture in poplar, Eucalyptus, birch, Zelkova schneideriana Hand.-Mazz., Acacia。
And, the described step condition that (4) middle high temperature sterilize processes is 121 DEG C of sterilizing 60min。
And, the pH of described step (4) middle ethyl sodium buffer solution is 4.5 5.0, and its compound method is: weigh sodium acetate, adding distilled water to dissolve, add glacial acetic acid, adding distilled water diluting to sodium acetate concentration is 0.5mol/L, the volumetric concentration of glacial acetic acid is 2.5%, and regulates pH value to 4.5 5.0。
And, the specification of described chip raw material is long 35mm × wide 10mm × thickness 5mm, washs 34 times by clear water。
And, the enzyme activity of described step (4) cellulase is 60.00FPU/mL, and the enzyme activity of hemicellulase is 85731.51U/mL。
And, described reactor is for sealing bag。
And, described step (4) in monosaccharide component be glucose or/and xylose。
The present invention obtain advantage and have the benefit effect that
1, this method adequately achieves effective utilization of this forestry waste of leaf wood brush wood; it is possible not only to effectively utilize forestry waste resource; also mitigate the pressure to environmental conservation; being benefit the nation and the people, meet National Sustainable Development Strategies and extremely urgent method, to the development of Chinese national economy, cleaning produces and environmental conservation is all significant。
2, this method realizes the Efficient Conversion of its Main chemical component by chemical Treatment, Mechanical Method process and cellulase treatment。Leaf wood slurry is after cellulase treatment, in substrate, cellulose is converted into the conversion ratio of glucose and can reach 93%, and the method can have high value added product for follow-up processing and utilization or production provides raw material basis, and the method cleans, simple, convenient, cost is reasonable。
Detailed description of the invention
Below in conjunction with embodiment, the present invention is further described；Following embodiment is illustrative, is not determinate, it is impossible to limit protection scope of the present invention with following embodiment。
The raw material used in the present invention, if no special instructions, is the commercially available prod of routine；The method used in the present invention, if no special instructions, is the conventional method of this area。
The pH of the ethyl sodium buffer solution used in the present invention is 4.5 5.0, and its compound method is: weigh sodium acetate (CH3COONa·3H2O), adding distilled water and dissolve, add glacial acetic acid, adding distilled water diluting to sodium acetate concentration is 0.5mol/L, and the volumetric concentration of glacial acetic acid is 2.5%, and regulates pH value to 4.5 5.0。
Embodiment 1
A kind of preprocess method realizing the efficient enzyme hydrolysis conversion of leaf wood brush wood, step is as follows:
(1) first leaf wood brush wood being cut into wood chip, obtain chip raw material, choose the wood chip that specification is about long 35mm × wide 10mm × thickness 5mm, clear water washs 34 times；
(2) leaf wood brush wood chip raw material is placed in boiling vessel, adds the H of 1.0% (relative to chip raw material absolute dry mass)2SO4Solid-to-liquid ratio is 1:10 (kg:L), boiling temperature is incubated 30min at the temperature of 160 DEG C, and in course of reaction, make the continuous circulated sprinkling of reactant liquor, so that leaf wood brush wood chip raw material and medicinal liquid fully react, after reaction terminates, pretreatment fluid and solid residue are easily separated, reacted leaf wood wood chip solid residue is washed to neutrality, stores for future use after air-dry, obtain wood chip after process；
(3) wood chip after the process after dilute sulfuric acid processes is placed in airtight reactor, is sequentially added into the Na of NaOH and 4.0% (relative to chip raw material absolute dry mass) of distilled water, 3.0% (relative to chip raw material absolute dry mass)2SO3, finally it is made into the concentration of 40% (relative to OD wood chip quality)。Being placed in water bath with thermostatic control by this closed reactor and carry out Chemical impregnation process, treatment temperature is 80 DEG C, processes time 30min；
(4) leaf wood brush wood wood chip is after the Chemical impregnation of above-mentioned steps processes, and processes through two sections of grindings: first paragraph is pressure grinding, and actual conditions is: temperature 120 DEG C, pressure 0.4MPa, starches dense 40%, grinding 5min；Second segment is normal pressure grinding, and actual conditions is: disk mill gap 0.10mm, starches dense 15%；
After (5) two sections of grindings process, leaf wood slurry is made into dense 3.0% (mass percent) of slurry, insulation 30min carries out disappearing latent in the water-bath (90 DEG C), then the leaf wood slurry after diving that disappears being discongested 10000 turns, leaf wood pulp fiber is through screening the fraction being collected by 0.15mm screen slot；It is about 30% by the leaf wood slurry recycling dehydration collected to concentration, obtains dehydration disposed slurry, then slurry collection is placed under 4 DEG C of conditions and stores and be used for follow-up biological enzyme hydrolysis；
(6) cellulase treatment: dehydration disposed slurry is carried out cellulase treatment to above-mentioned, before being processed, carries out high temperature sterilize process to container used, for instance 121 DEG C of sterilizing 60min；
Aseptically, ethyl sodium buffer solution is added in dehydration disposed slurry, the absolute dry mass of leaf wood slurry and ethyl sodium buffer solution ratio are 1:5 (kg:L), enzyme mixed liquor is added after being sufficiently stirred for, it is subsequently adding distilled water and makes leaf wood slurry, carry out enzyme constant temperature culture enzymolysis processing under aseptic condition, obtain glucose and xylose, it is achieved the efficient enzyme hydrolysis of leaf wood brush wood converts。
Concrete enzyme hydrolysis condition: 1. concentration of substrate 4.0%；2. reaction temperature 55 DEG C；3. response time 24 72 hours；4. pH4.8；5. rotating speed 180rpm；6. the consumption of cellulase is 24FPU/g substrate, and the consumption of hemicellulase is 60U/g substrate。
Testing result:
Under above-mentioned enzyme hydrolysis condition: after being hydrolyzed 72 hours, in hydrolyzed solution, the concentration of glucose is 18812mg/L, and the inversion rate of glucose of substrate is 83%；In hydrolyzed solution, the concentration of xylose is 3920mg/L, and the xylose rate of substrate is 77%。
Embodiment 2
A kind of preprocess method realizing the efficient enzyme hydrolysis conversion of leaf wood brush wood, step is as follows:
(1) first leaf wood brush wood being cut into wood chip, obtain chip raw material, choose the wood chip that specification is about long 35mm × wide 10mm × thickness 5mm, clear water washs 34 times；Described leaf wood is one or more the mixture in poplar, Eucalyptus, birch, Zelkova schneideriana Hand.-Mazz., Acacia；
(2) leaf wood brush wood chip raw material is placed in boiling vessel, adds the H of 1.0% (relative to chip raw material absolute dry mass)2SO4Solid-to-liquid ratio is 1:10 (kg:L), boiling temperature is incubated 60min at the temperature of 150 DEG C, and in course of reaction, make the continuous circulated sprinkling of reactant liquor, so that leaf wood brush wood chip raw material and medicinal liquid fully react, after reaction terminates, pretreatment fluid and solid residue are easily separated, reacted leaf wood wood chip solid residue is washed to neutrality, stores for future use after air-dry, obtain wood chip after process；
(3) wood chip after the process after dilute sulfuric acid processes is placed in airtight reactor, is sequentially added into the Na of NaOH and 4.0% (relative to chip raw material absolute dry mass) of distilled water, 3.0% (relative to chip raw material absolute dry mass)2SO3, finally it is made into the concentration of 40% (relative to OD wood chip quality)。Being placed in water bath with thermostatic control by this closed reactor and carry out Chemical impregnation process, treatment temperature is 80 DEG C, processes time 30min；
(4) leaf wood brush wood wood chip is after the Chemical impregnation of above-mentioned steps processes, and processes through two sections of grindings: first paragraph is pressure grinding, and actual conditions is: temperature 150 DEG C, pressure 0.4MPa, starches dense 40%, grinding 5min；Second segment is normal pressure grinding, and actual conditions is: disk mill gap 0.10mm, starches dense 15%；
After (5) two sections of grindings process, leaf wood slurry being made into slurry dense 3.0%, in water-bath (90 DEG C), insulation 30min carries out disappearing latent, then the leaf wood slurry after diving that disappears is discongested 10000 turns。Leaf wood pulp fiber is through screening the fraction being collected by 0.15mm screen slot, it is about 30% by the leaf wood slurry recycling dehydration collected to concentration, obtain dehydration disposed slurry, then slurry collection is placed under 4 DEG C of conditions and stores and be used for follow-up biological enzyme hydrolysis；
(6) cellulase treatment: dehydration disposed slurry is carried out cellulase treatment, before being processed, carries out high temperature sterilize process to container used；
Aseptically, ethyl sodium buffer solution is added in dehydration disposed slurry, the absolute dry mass of leaf wood slurry and ethyl sodium buffer solution ratio are 1:5 (kg:L), enzyme mixed liquor is added after being sufficiently stirred for, it is subsequently adding distilled water and makes leaf wood slurry, carry out enzyme constant temperature culture enzymolysis processing under aseptic condition, obtain glucose and xylose, it is achieved the efficient enzyme hydrolysis of leaf wood brush wood converts。
Concrete enzyme hydrolysis condition: 1. concentration of substrate 4.0%；2. reaction temperature 55 DEG C；3. response time 24 72 hours；4. pH4.8；5. rotating speed 180rpm；6. the consumption of cellulase is 20FPU/g substrate, and the consumption of hemicellulase is 60U/g substrate。
Testing result:
Under above-mentioned enzyme hydrolysis condition: after being hydrolyzed 72 hours, in hydrolyzed solution, the concentration of glucose is 19853mg/L, and the inversion rate of glucose of substrate is 82%；In hydrolyzed solution, the concentration of xylose is 4189mg/L, and the xylose rate of substrate is 63%。
Embodiment 3
A kind of preprocess method realizing the efficient enzyme hydrolysis conversion of leaf wood brush wood, step is as follows:
(1) first leaf wood brush wood being cut into wood chip, obtain chip raw material, choose the wood chip that specification is about long 35mm × wide 10mm × thickness 5mm, clear water washs 34 times；Described leaf wood is one or more the mixture in poplar, Eucalyptus, birch, Zelkova schneideriana Hand.-Mazz., Acacia；
(2) leaf wood brush wood chip raw material is placed in boiling vessel, adds the H of 1.0% (relative to chip raw material absolute dry mass)2SO4Solid-to-liquid ratio is 1:10 (kg:L), boiling temperature is incubated 40min at the temperature of 150 DEG C, and in course of reaction, make the continuous circulated sprinkling of reactant liquor, so that leaf wood brush wood chip raw material and medicinal liquid fully react, after reaction terminates, pretreatment fluid and solid residue are easily separated, reacted leaf wood wood chip solid residue is washed to neutrality, stores for future use after air-dry, obtain wood chip after process；
(3) wood chip after the process after dilute sulfuric acid processes is placed in airtight reactor, is sequentially added into the Na of NaOH and 4.0% (relative to chip raw material absolute dry mass) of distilled water, 3.0% (relative to chip raw material absolute dry mass)2SO3, finally it is made into the concentration of 40% (relative to OD wood chip quality)。Being placed in water bath with thermostatic control by this closed reactor and carry out Chemical impregnation process, treatment temperature is 80 DEG C, processes time 30min；
(4) leaf wood brush wood wood chip is after the Chemical impregnation of above-mentioned steps processes, and processes through two sections of grindings: first paragraph is pressure grinding, and actual conditions is: temperature 170 DEG C, pressure 0.4MPa, starches dense 40%, grinding 5min；Second segment is normal pressure grinding, and actual conditions is: disk mill gap 0.10mm, starches dense 15%；
After (5) two sections of grindings process, leaf wood slurry being made into slurry dense 3.0%, in water-bath (90 DEG C), insulation 30min carries out disappearing latent, then the leaf wood slurry after diving that disappears is discongested 10000 turns。Leaf wood pulp fiber is through screening the fraction being collected by 0.15mm screen slot。It is about 30% by the leaf wood slurry recycling dehydration collected to concentration, obtains dehydration disposed slurry, then slurry collection is placed under 4 DEG C of conditions and stores and be used for follow-up biological enzyme hydrolysis；
(6) cellulase treatment: dehydration disposed slurry is carried out cellulase treatment, before being processed, carries out high temperature sterilize process to container used；
Aseptically, ethyl sodium buffer solution is added in dehydration disposed slurry, the absolute dry mass of leaf wood slurry and ethyl sodium buffer solution ratio are 1:5 (kg:L), enzyme mixed liquor is added after being sufficiently stirred for, it is subsequently adding distilled water and makes leaf wood slurry, carry out enzyme constant temperature culture enzymolysis processing under aseptic condition, obtain glucose and xylose, it is achieved the efficient enzyme hydrolysis of leaf wood brush wood converts。
Concrete enzyme hydrolysis condition: 1. concentration of substrate 4.0%；2. reaction temperature 55 DEG C；3. response time 24 72 hours；4. pH4.8；5. rotating speed 180rpm；6. the consumption of cellulase is 20FPU/g substrate, and the consumption of hemicellulase is 60U/g substrate。
Testing result:
Under above-mentioned enzyme hydrolysis condition: after being hydrolyzed 72 hours, in hydrolyzed solution, the concentration of glucose is 21409mg/L, and the inversion rate of glucose of substrate is 93%；In hydrolyzed solution, the concentration of xylose is 4926mg/L, and the xylose rate of substrate is 55%。
Embodiment 4
A kind of preprocess method realizing the efficient enzyme hydrolysis conversion of leaf wood brush wood, step is as follows:
(1) first, leaf wood brush wood is cut into wood chip, obtains chip raw material；
(2) leaf wood brush wood chip raw material is placed in boiling vessel, adds the H relative to chip raw material absolute dry mass 0.5% 2.0%2SO4Solid-to-liquid ratio is 1:4 1:10 (kg:L), boiling temperature is incubated 20 90min at the temperature of 120 180 DEG C, and in course of reaction, make the continuous circulated sprinkling of reactant liquor, so that leaf wood brush wood chip raw material and medicinal liquid fully react, after reaction terminates, pretreatment fluid and solid residue are easily separated, reacted wood chip solid residue is washed with water to neutrality, stores for future use after air-dry, obtain wood chip after process；
(3) wood chip after the process after dilute sulfuric acid processes is carried out grinding process, disappear dive process, discongest process and processed after, obtain dehydration disposed slurry, then by dehydration disposed slurry collect and store for follow-up cellulase treatment in 4 DEG C of environment；
(4) cellulase treatment: dehydration disposed slurry is carried out cellulase treatment, before being processed, carries out high temperature sterilize process to container used；
Aseptically, ethyl sodium buffer solution is added in dehydration disposed slurry, the ratio (kg:L) of the absolute dry mass of dehydration disposed slurry and ethyl sodium buffer solution is 1:4 1:6, enzyme mixed liquor is added after being sufficiently stirred for, being subsequently adding water makes the mass concentration of leaf wood slurry be 1.0% 5.0%, carry out biological enzyme hydrolysis process under aseptic condition, monosaccharide component, it is achieved the efficient enzyme hydrolysis of leaf wood brush wood converts；
Wherein, the condition that described biological enzyme hydrolysis processes is: pH4.8, reaction temperature 40 55 DEG C, response time 10 90h, rotating speed 100 180rpm, cellulase consumption used is 10 30FPU/g substrates, and hemicellulose enzyme dosage is 20 80U/g substrates；
Above-mentioned steps concrete operation step (3) is as follows:
1. wood chip after the process after dilute sulfuric acid processes is placed in airtight reactor, is sequentially added into distilled water, the NaOH relative to chip raw material absolute dry mass 1.0% 5.0% and the Na relative to chip raw material absolute dry mass 1.0% 5.0%2SO3, finally it being made into the mass percentage concentration relative to chip raw material absolute dry mass 20% 50%, this closed reactor is placed in water bath with thermostatic control and carries out Chemical impregnation process, treatment temperature is 60 90 DEG C, processes time 20 40min；
2. after Chemical impregnation processes, after process, wood chip processes then through two sections of grindings: first paragraph is pressure grinding, and actual conditions is: temperature 120 180 DEG C, pressure 0.1 0.5MPa, starches dense 10% 40% (mass percents), grinding 2 10min；Second segment is normal pressure grinding, and actual conditions is: disk mill gap 0.08 0.10mm, starches dense 10% 30% (mass percents), obtains leaf wood slurry；
3. after two sections of grindings process, leaf wood slurry is made into dense 1.0% 5.0% (mass percents) of slurry, under 80 90 DEG C of conditions, it is incubated 10 30min carries out the process of diving that disappears, then the leaf wood slurry after diving that disappears is discongested 10,000 30000 turns, obtain leaf wood pulp fiber；Leaf wood pulp fiber is through screening the fraction being collected by 0.15mm screen slot, it is 20%-40% by the leaf wood slurry recycling dehydration collected to mass percentage concentration, obtain dehydration disposed slurry, then dehydration disposed slurry collected and store for follow-up cellulase treatment in 4 DEG C of environment；
Or, described step concrete operation step (3) is as follows:
1. being placed in grinding device by wood chip after the process after dilute sulfuric acid processes and carry out grinding process, actual conditions is: disk mill gap 0.08 0.10mm, starches dense 10% 30% (mass percents), obtains leaf wood slurry；
2. after grinding processes, leaf wood slurry is made into dense 1.0% 5.0% (mass percents) of slurry, under 80 90 DEG C of conditions, it is incubated 10 30min carries out the process of diving that disappears, then the leaf wood slurry after diving that disappears is discongested 10,000 30000 turns, obtain leaf wood pulp fiber；Leaf wood pulp fiber is through screening the fraction being collected by 0.15mm screen slot, it is 20%-40% by the leaf wood slurry recycling dehydration collected to mass percentage concentration, obtain dehydration disposed slurry, then dehydration disposed slurry collected and store for follow-up cellulase treatment in 4 DEG C of environment；
The described step condition that (4) middle high temperature sterilize processes is 121 DEG C of sterilizing 60min。
The pH of described step (4) middle ethyl sodium buffer solution is 4.5 5.0, and its compound method is: weigh sodium acetate, adds distilled water and dissolves, add glacial acetic acid, adding distilled water diluting to sodium acetate concentration is 0.5mol/L, and the volumetric concentration of glacial acetic acid is 2.5%, and regulates pH value to 4.5 5.0。
The specification of described chip raw material is long 35mm × wide 10mm × thickness 5mm, washs 34 times by clear water
The enzyme activity of described step (4) cellulase is 60.00FPU/mL, and the enzyme activity of hemicellulase is 85731.51U/mL。
Described step (4) in monosaccharide component be glucose or/and xylose。
Testing result:
Testing result is with embodiment 2。
Claims (9)
1. one kind realizes the preprocess method that the efficient enzyme hydrolysis of leaf wood brush wood converts, it is characterised in that: step is as follows:
(1) first, leaf wood brush wood is cut into wood chip, obtains chip raw material；
(2) leaf wood brush wood chip raw material is placed in boiling vessel, adds the H relative to chip raw material absolute dry mass 0.5% 2.0%2SO4Solid-to-liquid ratio is 1:4 1:10 (kg:L), boiling temperature is incubated 20 90min at the temperature of 120 180 DEG C, and in course of reaction, make the continuous circulated sprinkling of reactant liquor, so that leaf wood brush wood chip raw material and medicinal liquid fully react, after reaction terminates, pretreatment fluid and solid residue are easily separated, reacted wood chip solid residue is washed with water to neutrality, stores for future use after air-dry, obtain wood chip after process；
(3) wood chip after the process after dilute sulfuric acid processes is carried out grinding process, disappear dive process, discongest process and processed after, obtain dehydration disposed slurry, then by dehydration disposed slurry collect and store for follow-up cellulase treatment in 4 DEG C of environment；
(4) cellulase treatment: dehydration disposed slurry is carried out cellulase treatment, before being processed, carries out high temperature sterilize process to container used；
Aseptically, ethyl sodium buffer solution is added in dehydration disposed slurry, the ratio (kg:L) of the absolute dry mass of dehydration disposed slurry and ethyl sodium buffer solution is 1:4 1:6, enzyme mixed liquor is added after being sufficiently stirred for, being subsequently adding water makes the mass concentration of leaf wood slurry be 1.0% 5.0%, carry out biological enzyme hydrolysis process under aseptic condition, monosaccharide component, it is achieved the efficient enzyme hydrolysis of leaf wood brush wood converts；
Wherein, the condition that described biological enzyme hydrolysis processes is: pH4.8, reaction temperature 40 55 DEG C, response time 10 90h, rotating speed 100 180rpm, cellulase consumption used is 10 30FPU/g substrates, and hemicellulose enzyme dosage is 20 80U/g substrates。
2. the preprocess method realizing the efficient enzyme hydrolysis conversion of leaf wood brush wood according to claim 1, it is characterised in that: described step concrete operation step (3) is as follows:
1. wood chip after the process after dilute sulfuric acid processes is placed in airtight reactor, is sequentially added into distilled water, the NaOH relative to chip raw material absolute dry mass 1.0% 5.0% and the Na relative to chip raw material absolute dry mass 1.0% 5.0%2SO3, finally it being made into the mass percentage concentration relative to chip raw material absolute dry mass 20% 50%, this closed reactor is placed in water bath with thermostatic control and carries out Chemical impregnation process, treatment temperature is 60 90 DEG C, processes time 20 40min；
2. after Chemical impregnation processes, after process, wood chip processes then through two sections of grindings: first paragraph is pressure grinding, and actual conditions is: temperature 120 180 DEG C, pressure 0.1 0.5MPa, starches dense 10% 40% (mass percents), grinding 2 10min；Second segment is normal pressure grinding, and actual conditions is: disk mill gap 0.08 0.10mm, starches dense 10% 30% (mass percents), obtains leaf wood slurry；
3. after two sections of grindings process, leaf wood slurry is made into dense 1.0% 5.0% (mass percents) of slurry, under 80 90 DEG C of conditions, it is incubated 10 30min carries out the process of diving that disappears, then the leaf wood slurry after diving that disappears is discongested 10,000 30000 turns, obtain leaf wood pulp fiber；Leaf wood pulp fiber is through screening the fraction being collected by 0.15mm screen slot, it is 20%-40% by the leaf wood slurry recycling dehydration collected to mass percentage concentration, obtain dehydration disposed slurry, then dehydration disposed slurry collected and store for follow-up cellulase treatment in 4 DEG C of environment；
Or, described step concrete operation step (3) is as follows:
1. being placed in grinding device by wood chip after the process after dilute sulfuric acid processes and carry out grinding process, actual conditions is: disk mill gap 0.08 0.10mm, starches dense 10% 30% (mass percents), obtains leaf wood slurry；
2. after grinding processes, leaf wood slurry is made into dense 1.0% 5.0% (mass percents) of slurry, under 80 90 DEG C of conditions, it is incubated 10 30min carries out the process of diving that disappears, then the leaf wood slurry after diving that disappears is discongested 10,000 30000 turns, obtain leaf wood pulp fiber；Leaf wood pulp fiber is through screening the fraction being collected by 0.15mm screen slot, it is 20%-40% by the leaf wood slurry recycling dehydration collected to mass percentage concentration, obtain dehydration disposed slurry, then dehydration disposed slurry collected and store for follow-up cellulase treatment in 4 DEG C of environment。
3. the preprocess method realizing the efficient enzyme hydrolysis conversion of leaf wood brush wood according to claim 1 and 2, it is characterised in that: described leaf wood is one or more the mixture in poplar, Eucalyptus, birch, Zelkova schneideriana Hand.-Mazz., Acacia。
4. the preprocess method realizing the efficient enzyme hydrolysis conversion of leaf wood brush wood according to claim 1 and 2, it is characterised in that: the described step condition that (4) middle high temperature sterilize processes is 121 DEG C of sterilizing 60min。
5. the preprocess method realizing the efficient enzyme hydrolysis conversion of leaf wood brush wood according to claim 1 and 2, it is characterized in that: the pH of described step (4) middle ethyl sodium buffer solution is 4.5 5.0, its compound method is: weigh sodium acetate, add distilled water to dissolve, add glacial acetic acid, adding distilled water diluting to sodium acetate concentration is 0.5mol/L, and the volumetric concentration of glacial acetic acid is 2.5%, and regulates pH value to 4.5 5.0。
6. the preprocess method realizing the efficient enzyme hydrolysis conversion of leaf wood brush wood according to claim 1 and 2, it is characterised in that: the specification of described chip raw material is long 35mm × wide 10mm × thickness 5mm, washs 34 times by clear water。
7. the preprocess method realizing the efficient enzyme hydrolysis conversion of leaf wood brush wood according to claim 1 and 2, it is characterised in that: the enzyme activity of described step (4) cellulase is 60.00FPU/mL, and the enzyme activity of hemicellulase is 85731.51U/mL。
8. the preprocess method realizing the efficient enzyme hydrolysis conversion of leaf wood brush wood according to claim 2, it is characterised in that: described reactor is for sealing bag。
9. according to claim 1 and 2 realize the preprocess method that the efficient enzyme hydrolysis of leaf wood brush wood converts, it is characterised in that: described step (4) in monosaccharide component be glucose or/and xylose。
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