CN110257453A - A kind of preprocess method for improving fibrous raw material biological enzymolysis conversion ratio and preparing fermentable sugars - Google Patents
A kind of preprocess method for improving fibrous raw material biological enzymolysis conversion ratio and preparing fermentable sugars Download PDFInfo
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- CN110257453A CN110257453A CN201910451353.6A CN201910451353A CN110257453A CN 110257453 A CN110257453 A CN 110257453A CN 201910451353 A CN201910451353 A CN 201910451353A CN 110257453 A CN110257453 A CN 110257453A
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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
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
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Abstract
The invention proposes the preprocess methods that a kind of raising fibrous raw material biological enzymolysis conversion ratio prepares fermentable sugars, are pre-processed using Cold pretreatment technology to lignocellulosic material, effectively abolish the inhibiting factor for restricting raw cellulose enzyme hydrolysis.This method comprises: the size degradation of (1) fibrous raw material；(2) pretreatment of fines: pretreatment process repeated several times, addition chemicals usage every time are as follows: hydrogen peroxide use 3-20%, sodium hydroxide concentration 3-20%, amount of sodium silicate 1-5%, DTPA dosage 0.3-5%, above-mentioned chemicals usage are the mass percent relative to over dry material, entire pretreatment system solid masses concentration is 5-30%, solvent is water and water-miscible organic solvent, and 70 DEG C -150 DEG C of reaction temperature, the reaction time is 30-180 minutes.To the biggish bamboo class raw material of enzyme hydrolysis antagonism, pretreatment temperature is no more than 95 DEG C, can be realized enzyme hydrolysis section inversion rate of glucose 90%, and total glucose rate of recovery is close to 90%.
Description
Technical field
The invention belongs to the preparation field of fermentable sugars, in particular to a kind of raising fibrous raw material biological enzymolysis conversion ratio system
The preprocess method of standby fermentable sugars.
Background technique
Lignocellulose raw material is mainly made of cellulose, hemicellulose and lignin, and wherein hemicellulose and lignin are tight
It is close to wrap cellulose, form the physics and chemical structure of wood fiber biomass densification.Therefore, lignocellulose raw material is utilized
Biorefinery is carried out firstly the need of being pre-processed, raw material physicalchemical structure is destroyed, helps cellulase and cell wall internal layer
Cellulose contact, enzyme hydrolysis (Tekin K, Hao N, Karagoz S, Ragauskas A J.Ethanol:A is carried out to it
Promising Green Solvent for the Deconstruction of Lignocellulose.ChemSusChem,
2018,11:3559-3575.Min D,Li Q,Chiang V,Jameel H,Chang H M,Lucia L.The
influence of lignin-carbohydrate complexes on the cellulase-mediated
saccharification II:transgenic hybrid poplars(Populus nigra L.and Populus
maximowiczii A.).Fuel,2014,116:56-62)。
Conventional preprocess method include dilute acid pretreatment, hydrothermal pretreatment, alkaline process pretreatment, steam blasting pretreatment etc. all
The structure that raw material can effectively be destroyed reaches higher cellulose hydrolyzation yield.Wherein, to conventional wood raw material, diluted acid is located in advance
Reason reaction temperature at least needs 160 DEG C, and sour dosage 1%, enzymatic hydrolysis section glucose yield is no more than 80%, and total glucose rate of recovery is not
More than 60%, (Lv Jiaqing, Bi Yanjin, Yan Xixi, et al. dilute sulfuric acid hydrolysis pretreatment are to ryegrass pulping stock waste residue enzyme hydrolysis
Influence [J] Nanjing Forestry University's journal (natural science edition) of saccharification, 2015 (4): 137-141.)；Hydrothermal pretreatment reaction temperature
Degree at least needs 180 DEG C, and enzymatic hydrolysis section glucose yield is no more than 80%, total glucose rate of recovery be no more than 90% [Huang C,
Wu X,Huang Y,Lai C,Li X,Yong Q.Prewashing enhances the liquid hot water
pretreatment efficiency of waste wheat straw with high free ash
content.Bioresoure Technology,2016,219:583-588]；Alkaline process pretreatment reaction temperature at least needs 120
DEG C, base amount 2-4%, enzymatic hydrolysis section glucose yield be no more than 70%, total glucose rate of recovery be no more than 70% (Deng Hui, Li Chun,
Li Fei, et al. cotton stalk saccharification oxygenation pretreatment condition optimizing [J] Journal of Agricultural Engineering, 2009 (1): 208-212.)；It steams
Steam explosion breaks pretreatment reaction temperature and at least needs 220 DEG C, and enzymatic hydrolysis section glucose yield is no more than 80%, and total glucose rate of recovery is not
More than 70% (research [J] of Luo Peng, Liu Zhong, Yang Chuanmin, et al. steam blasting wheat straw simultaneous saccharification and fermentation ethanol conversion
Learn engineering, 2007,35 (12): 42-45.).
Compared with conventional lignocellulose raw material, the degree of lignification of bamboo wood is higher, bigger using difficulty.Has scholar to bamboo
Material is pre-processed, although discovery acid system pretreatment can remove wherein most hemicellulose, pretreated material lignin
Content is still higher, thus pretreated material enzyme hydrolysis yield is relatively low；And delignified preprocess method can then effectively improve
The enzyme hydrolysis yield of pretreated material.And the bamboo particle pretreatment method of current mainstream is made a general survey of, it is seen that at present about bamboo wood
Pretreatment is carried out at higher temperature (being greater than 140 DEG C) and biggish alkali charge (being greater than 4%), fine in preprocessing process
The problems such as dimension procatarxis is partly dissolved for peeling reaction, causes the glucose rate of recovery low (less than 50%) [Huang C, Ma
J,Liang C,Li X,Yong Q.Influence of sulfur dioxide-ethanol-water pretreatment
on the physicochemical properties and enzymatic digestibility of bamboo
residues.Bioresource Technology,2018,263:17-24]。
Summary of the invention
Goal of the invention: existing fibrous raw material preconditioning technique reaction temperature is higher, the high requirements on the equipment, operating cost
Height, the pretreatment section glucose rate of recovery is not high, and total glucose rate of recovery is not high, to solve the above problems, the invention proposes one
The preprocess method that kind raising fibrous raw material biological enzymolysis conversion ratio prepares fermentable sugars can using technical method of the invention
Under the pretreatment temperature no more than 95 DEG C, so that it may which total glucose rate of recovery of fibrous raw material is increased to 90% or so.
Technical solution: to realize the above-mentioned technical purpose, raising fibrous raw material biological enzymolysis conversion ratio preparation of the invention can
The preprocess method of sugar fermentation, includes the following steps:
(1) size degradation of fibrous raw material: fibrous raw material is washed, impregnate, after the processing of mechanical presses equipment at filament bulk, so
Fines is obtained after the processing of normal pressure mill afterwards；
(2) pretreatment of fines: pretreatment process repeated several times, it is preferable that repeat 1~5 time, certain duplicate number
It is The more the better, chemicals usage is added every time are as follows: hydrogen peroxide use 3-20%, sodium hydroxide concentration 3-20%, sodium metasilicate are used
1-5% is measured, DTPA dosage 0.3-5%, above-mentioned chemicals usage is the mass percent relative to over dry material, entire pre- place
Reason system solid masses concentration be 5-30%, solvent be water and water-miscible organic solvent, 70 DEG C -150 DEG C of reaction temperature,
Reaction time is 30-180 minutes.
Preferably, in step (1) after soak at room temperature 12 hours, at filament bulk after the processing of mechanical presses equipment, normal
The fines of size 1-10mm is obtained after the processing of platen mill.
Specifically, the solvent is methanol, ethyl alcohol, acetone, tetrahydrofuran, fourth sulfone, dioxane, ethylene glycol, 1,3 dioxy
Any one or a few arbitrary proportion mixed solution in heterocycle pentane.
In step (2), all chemicals and solvent once put into reactor, are uniformly mixed with fines and are heated to set
Insulation reaction after temperature, reaching and material taking-up is washed to filtrate pH by distillation in Buchner funnel after designing the reaction time is 7.
Specifically, the fibrous raw material be bamboo wood, it is poplar, wheat straw, Eucalyptus, moso bamboo, cotton stalk, any one in corncob
Kind.
Particularly, processing method of the invention has excellent treatment effect for bamboo wood.
The utility model has the advantages that the present invention (is not less than 70 degrees Celsius, and can obtain at 95 DEG C with respect to the lower temperature of prior art
Excellent effect) under conditions of, the physical chemistry barrier of constraints on fiber element efficiently saccharifying can be broken through, realizes that enzyme hydrolysis section is higher
Glucose yield (90%), while in pretreatment section keeping higher cellulose conversion ratio (total glucose rate of recovery is more than
80%), fibrous raw material especially high to lignin content, property have the effect of for traditional treatment method it is excellent, it is especially right
In bamboo wood, conventional method total reducing sugar conversion ratio only has 50% or so, and this method can achieve 90%, and the method for the present invention reaction condition
Relatively mild, not high to equipment requirement, chemicals used is conventional dose, and operation cost is low.
Specific embodiment
The invention proposes the preprocess method that a kind of raising fibrous raw material biological enzymolysis conversion ratio prepares fermentable sugars, packets
Include following steps:
(1) size degradation of fibrous raw material: fibrous raw material is washed, impregnate, after the processing of mechanical presses equipment at filament bulk, so
Fines is obtained after the processing of normal pressure mill afterwards；
(2) pretreatment of fines: pretreatment process repeated several times add chemicals usage every time are as follows: hydrogen peroxide is used
3-20%, sodium hydroxide concentration 3-20%, amount of sodium silicate 1-5%, DTPA dosage 0.3-5% are measured, above-mentioned chemicals usage is equal
For the mass percent relative to over dry material, entire pretreatment system solid masses concentration is 5-30%, and solvent is water and energy
Organic solvent miscible with water, such as methanol, ethyl alcohol, acetone, tetrahydrofuran, fourth sulfone, dioxane, ethylene glycol, 1,3 dioxane
Any one or a few arbitrary proportion mixed solution in pentane, 70 DEG C -150 DEG C of reaction temperature, reaction time 30-180
Minute.All chemicals and solvent once put into reactor, are uniformly mixed after being heated to set temperature and are kept the temperature instead with fines
It answers, reaching and material taking-up is washed to filtrate pH by distillation in Buchner funnel after designing the reaction time is 7.
Below by specific embodiment, the present invention will be described in detail.
Wherein, in following embodiments the step of saccharification of cellulose are as follows:
The material obtained after the processing of over dry weight 2.5g is weighed in 150mL enzymatic hydrolysis bottle, 1mol/L lemon acid buffering is added
Liquid 2.5mL, is added cellulase by cellulase dosage 25FPU/g cellulose, and finally supplementing distilled water makes enzymatic hydrolysis system volume
For 50mL；Enzymatic hydrolysis bottle is placed in 150rpm, digests 72h in 50 DEG C of constant-temperature tables, after enzymatic hydrolysis, takes enzymatic hydrolysis sample, in
It is centrifuged under 10000rpm, takes supernatant to measure concentration of glucose therein and cellobiose concentration, and calculate enzyme hydrolysis yield.
Wherein, section glucose yield (%)=[(fiber two in concentration of glucose g/L*0.9+ hydrolyzate in hydrolyzate is digested
Sugared concentration g/L*0.95) * 0.05/ (substrate over dry weight g* content of cellulose)] * 100%, 0.9 is glucose and cellulose in formula
Conversion coefficient, 0.95 be cellobiose and cellulose conversion coefficient, 0.05 be hydrolyzation system volume；
Total glucose yield (%)=[(cellobiose concentration g/ in concentration of glucose g/L*0.9+ hydrolyzate in hydrolyzate
) * 0.05/ L*0.95 (content of cellulose in raw material over dry weight g* raw material)] * 100%, 0.9 is glucose and cellulose in formula
Conversion coefficient, 0.95 is the conversion coefficient of cellobiose and cellulose, and 0.05 is hydrolyzation system volume.
Influence of the 1 different pretreatments process of embodiment to fibrous material inversion rate of glucose.
Use bamboo wood for raw material in following examples, content of cellulose 43.14%, specific steps are as follows: raw material is by conventional
After size degradation, the fines for being equivalent to 10g oven dry weight is weighed, all kinds of chemicals are uniformly mixed by 1 dosage of according to the form below with fines, with steaming
Distilled water is put into closed reactor after being adjusted to system concentration 20%, insulation reaction 60min, gains after being heated to 95 degrees Celsius
Material is spare after cleaning.Above-mentioned pretreatment reaction process is repeated if experimental design needs multistage to handle.Resulting material is by normal
Rule saccharification reaction, obtains corresponding inversion rate of glucose, rate of recovery data see the table below.
Table 1
As seen from the above table, do not only have 13% by pretreated material inversion rate of glucose, and ethyl alcohol is used alone
(1), hydrogen peroxide (2), sodium hydroxide (3), are pre-processed at 95 DEG C, glucolase solution yield and total glucose rate of recovery
Lower, using aqueous alkaline hydrogen peroxide treatment (4,5), total glucose rate of recovery is no more than 73%, using multistage ethyl alcohol-alkali
Property hydrogen peroxide pretreatment (7,8,9), enzymatic hydrolysis glucose yield be greater than 90%, total glucose rate of recovery is 90% or so.I.e. originally
Effect is best when technology uses organic solvent-hydrogen peroxide-sodium hydroxide.
Influence of the 2 different pretreatments process of embodiment to fibrous material inversion rate of glucose.
Use bamboo wood for raw material in following examples, content of cellulose 43.14%, specific steps are as follows: raw material is by conventional
After size degradation, the fines for being equivalent to 10g oven dry weight is weighed, all kinds of chemicals are uniformly mixed by according to the form below dosage with fines, with steaming
Distilled water is put into closed reactor after being adjusted to system concentration 20%, is heated to insulation reaction 60min after setting, and resulting material is washed
It is spare after net.Above-mentioned pretreatment reaction process is repeated if experimental design needs multistage to handle.Resulting material is by conventional sugar
Change reaction, obtains corresponding inversion rate of glucose, rate of recovery data are shown in Table 2.
Table 2
Embodiment 3
Use poplar for raw material, content of cellulose 45.43% handles material by the operating procedure of example one, wherein in advance
Treatment conditions: the material for being equivalent to 10g oven dry weight, hydrogen peroxide use 8%, sodium hydroxide concentration 9%, sodium metasilicate use are weighed
Measure 2%, DTPA dosage 0.5%, medical fluid point three Duan Tianjia, every section of processing uses dioxane dosage 7g, and 95 DEG C of temperature, reactant
It is concentration 20%, gained digests section glucose yield 94.53%, total glucose rate of recovery 91.47%.
Embodiment 4
Use wheat straw for raw material, content of cellulose 35.64% handles material by the operating procedure of example one, wherein in advance
Treatment conditions: the material for being equivalent to 10g oven dry weight, hydrogen peroxide use 10%, sodium hydroxide concentration 6%, sodium metasilicate are weighed
Dosage 2%, DTPA dosage 0.5%, medical fluid are divided to two sections of additions, and every section of processing uses acetone dosage 4g, and 95 DEG C of temperature, reaction system
Concentration 30%, gained digest section glucose yield 85.41%, total glucose rate of recovery 81.54%.
Embodiment 5
Use Eucalyptus for raw material, content of cellulose 42.63% handles material by the operating procedure of example one, wherein in advance
Treatment conditions: the material for being equivalent to 10g oven dry weight, hydrogen peroxide use 8%, sodium hydroxide concentration 4.5%, sodium metasilicate are weighed
Dosage 2%, DTPA dosage 0.5%, a medical fluid point Duan Tianjia, every section of processing use methanol usage 60%, and 95 DEG C of temperature, concentration
20%, gained digests section glucose yield 93.69%, total glucose rate of recovery 90.72%.
Embodiment 6
Use moso bamboo for raw material, content of cellulose 43.14% handles material by the operating procedure of example one, wherein in advance
Treatment conditions: the material for being equivalent to 10g oven dry weight, hydrogen peroxide use 15%, sodium hydroxide concentration 10%, sodium metasilicate are weighed
Dosage 2%, DTPA dosage 0.5%, medical fluid point three Duan Tianjia, every section of processing use fourth sulfone dosage 90%, and 95 DEG C of temperature, concentration
10%, gained digests section glucose yield 94.69%, total glucose rate of recovery 89.58%.
Embodiment 7
Use cotton stalk for raw material, content of cellulose 31.69% handles material by the operating procedure of example one, wherein in advance
Treatment conditions: the material for being equivalent to 10g oven dry weight, hydrogen peroxide use 9%, sodium hydroxide concentration 6%, sodium metasilicate use are weighed
Measure 2%, DTPA dosage 0.5%, a medical fluid point Duan Tianjia, every section of processing uses ethylene glycol dosage 80%, and 95 DEG C of temperature, concentration
5%, gained digests section glucose yield 88.53%, total glucose rate of recovery 81.36%.
Embodiment 8
Use corncob for raw material, content of cellulose 28.57% handles material by the operating procedure of example one, wherein
Pretreatment condition: the material for being equivalent to 10g oven dry weight, hydrogen peroxide use 9%, sodium hydroxide concentration 5%, sodium metasilicate are weighed
Dosage 2%, DTPA dosage 0.5%, a medical fluid point Duan Tianjia, every section of processing uses tetrahydrofuran dosage 50%, 95 DEG C of temperature, dense
Degree 20%, gained digest section glucose yield 89.87%, total glucose rate of recovery 84.69%.
Claims (6)
1. a kind of preprocess method for improving fibrous raw material biological enzymolysis conversion ratio and preparing fermentable sugars, which is characterized in that including
Following steps:
(1) size degradation of fibrous raw material: fibrous raw material is washed, impregnate, after the processing of mechanical presses equipment at filament bulk, then exist
Fines is obtained after the processing of normal pressure mill；
(2) pretreatment of fines: pretreatment process repeated several times add chemicals usage every time are as follows: hydrogen peroxide use 3-
20%, sodium hydroxide concentration 3-20%, amount of sodium silicate 1-5%, DTPA dosage 0.3-5%, above-mentioned chemicals usage be relative to
The mass percent of over dry material, entire pretreatment system solid masses concentration are 5-30%, and solvent is water and water-miscible
Organic solvent, 70 DEG C -150 DEG C of reaction temperature, the reaction time is 30-180 minutes.
2. preprocess method according to claim 1, which is characterized in that in step (1), obtained after the processing of normal pressure mill
Fines having a size of 1-10mm.
3. preprocess method according to claim 1, which is characterized in that the solvent is methanol, ethyl alcohol, acetone, tetrahydro
Furans, fourth sulfone, dioxane, ethylene glycol, any one or a few the arbitrary proportion mixing in 1,3 dioxolane are molten
Liquid.
4. preprocess method according to claim 1, which is characterized in that in step (2), all chemicals and solvent one
Secondary property puts into reactor, and insulation reaction after being heated to set temperature is uniformly mixed with fines, reaches object after designing the reaction time
It is 7 that material, which takes out and is washed to filtrate pH by distillation in Buchner funnel,.
5. preprocess method according to claim 1, which is characterized in that the fibrous raw material be bamboo wood, poplar, wheat straw,
Eucalyptus, moso bamboo, cotton stalk, any one in corncob.
6. preprocess method according to claim 1, which is characterized in that the fibrous raw material is bamboo wood.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115029386A (en) * | 2022-05-07 | 2022-09-09 | 中国林业科学研究院林产化学工业研究所 | Method for improving yield and productivity of methane prepared by solid-state anaerobic fermentation of agricultural and forestry residues |
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| CN115029386B (en) * | 2022-05-07 | 2023-09-01 | 中国林业科学研究院林产化学工业研究所 | Method for improving yield and productivity of methane prepared by solid anaerobic fermentation of agriculture and forestry residues |
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