CN109371077A - A method of biological syrup is prepared using biomass - Google Patents
A method of biological syrup is prepared using biomass Download PDFInfo
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- CN109371077A CN109371077A CN201811402058.3A CN201811402058A CN109371077A CN 109371077 A CN109371077 A CN 109371077A CN 201811402058 A CN201811402058 A CN 201811402058A CN 109371077 A CN109371077 A CN 109371077A
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- 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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Abstract
The present invention relates to a kind of methods for preparing biological syrup using biomass, using biological material as raw material, the biomass material of certain mesh number will be obtained by screening after biological material progress pretreatment early period, biomass material is dried, then it weighs a certain amount of biomass material and prepares certain density suspension, adjust the pH value of suspension, keep certain temperature, a certain amount of pectase and ferulic acid lipase combination enzyme is added, and it is stirred under certain revolving speed, enzymatic hydrolysis a period of time, pH value is adjusted again and keeps certain temperature, add a certain amount of fine group synthase for tieing up plain enzyme ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase, add a certain amount of L-AA, certain time is digested again.The present invention realizes the time of the Jian Shao ﹑ enzymatic activity maintenance of the Suo Duan ﹑ enzyme additive amount of enzymolysis time for a long time and the Zai Li of enzyme reduces cost etc. Yong ﹑, obtains the syrup enzymolysis liquid of high concentration.
Description
Technical field
The invention belongs to biological materials to utilize technical field, in particular to a kind of to prepare biological syrup using biomass
Method.
Background technique
Lignocellulosic is from a wealth of sources, cheap, is the desirable feedstock for producing alcohol fuel, and main component includes
30% ~ 40% cellulose, 20% ~ 30% hemicellulose and 20% ~ 30% lignin.Biological material such as corn stover, corn
Core, sugarcane, wheat, contains a large amount of lignocellulosic in sugar grass at agriculture and forestry organic waste material.
Lignocellulosic prepares cellulosic ethanol and generally includes the units operation such as pretreatment, hydrolysis, fermentation, distillation at present,
It is vital link in cellulosic ethanol refining process that wherein cellulose, which is hydrolyzed to fermentable sugars,.Currently, lignocellulosic
Degradation mainly have chemical method and enzymatic hydrolysis.Compared with chemical method hydrolysis, enzymatic hydrolysis mild condition, equipment is simple,
Low energy consumption, while having the characteristics that by-product is few, environmental-friendly, in widespread attention and make substantial progress.Due to wooden fibre
Cellulose in dimension element mainly exists with crystalline state, it is therefore desirable to which it is quite important that the enzyme of high activity carrys out hydrocellulose.But mesh
Preceding existing fiber enzyme activity is low, and the component of cellulase is single, and consumption enzyme amount is big, and enzymolysis time is long, and fermentation costs are high, and its
Enzymolysis efficiency and conversion ratio under acid condition is generally relatively low, seriously limits lignocellulosic answering in bio-ethanol production
With.
It is existing more to the method for lignocellulosic enzymatic hydrolysis, it is disclosed in the Chinese patent of notification number CN107916284A
A kind of " digesting saccharification technology by the lignocellulosic of medium of polyethylene glycol/aqueous mixtures ", using lignocellulosic as raw material, gathers
Ethylene glycol replaces part water, and the enzymatic saccharification process of lignocellulosic is used for using polyethylene glycol/aqueous mixtures as enzymatic hydrolysis medium
In, both reduced the dosage of water while or so that the viscosity of reaction volume is effectively controlled, lignocellulosic digest efficiency not by
It influences, the concentration of reduced sugar of obtained lignocellulosic enzyme hydrolyzate is improved.But there is also following major defects for this method:
(1) additive amount of the cellulase used is larger, at high cost；(2) enzymolysis time is more than 100h, and enzymolysis efficiency is lower；(3) it digests
Obtain concentration of glucose 35g/l ~ 65g/l, concentration of glucose is relatively low, has a certain impact to subsequent fermentation.
Summary of the invention
Technical problem to be solved by the present invention lies in, a kind of method preparing biological syrup using biomass is provided, with
Biomass is raw material, the syrup of high-content is prepared by using two step enzymatic hydrolysis, the first step is by using pectase and orizanol
Enzyme group synthase digests its cell wall, carrys out the pectic substance in fast degradation cell wall, reduces the viscosity of its material；The
Two steps tie up plain enzyme ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase group synthase and L-AA by fine, to fiber
Element carries out effectively hydrolyzing and prepares syrup, and cellulase may be implemented and preferably contact substrate, so that the Suo Duan ﹑ enzyme of enzymolysis time adds
The time that the Jian Shao ﹑ enzymatic activity of dosage maintains is for a long time and the Zai Li of enzyme reduces cost etc. Yong ﹑, obtains the syrup enzymatic hydrolysis of high concentration
Liquid.
The invention is realized in this way a kind of method for preparing biological syrup using biomass is provided, with biological material
For raw material, the biomass material of certain mesh number will be obtained by screening after biological material progress pretreatment early period, by biomass
Material is dried, and then weighs a certain amount of biomass material and prepares certain density suspension, adjusts suspension
PH value keeps certain temperature, a certain amount of pectase and ferulic acid lipase combination enzyme is added, and carry out under certain revolving speed
Stirring, enzymatic hydrolysis a period of time, then adjust pH value and keep certain temperature, it is more to add a certain amount of Xian dimension Su Mei ﹑ dissolubility
The group synthase of sugared monooxygenase (LPMO) ﹑ catalase, adds a certain amount of L-AA, then digest certain time,
Then it is filtered by filter paper, the solid-liquid in enzymolysis liquid is separated, enzymolysis liquid is obtained, then by enzymolysis liquid in Rotary Evaporators
In be concentrated in vacuo, obtain biological syrup.
Further, the method for preparing biological syrup using biomass includes the following steps:
Step 1: the pretreatment of biological material: biological material is used steam pre-treatment, dilute acid pretreatment, wet oxidation, use
Wet explosive pretreatment, Biological Pretreatment, the supercritical CO of organic solvent2Pretreatment, overcritical H2O is pre-processed, ozone pre-processes,
Ionic liquid pretreatment, ultrasonic pretreatment, Microwave Pretreatment, γ radiate any one mode in pretreatment and are pre-processed；
Step 2: biological material is dried and screening process: by the dry 48h of pretreated biological material, then
40 mesh ~ 60 mesh biomass material is sieved with sieve；
Step 3: preparing the suspension of biomass material: weighing biomass material 40g, be configured to concentration of substrate with deionized water
15% ~ 30% biomass suspension；
Step 4: adjusting the pH value and temperature of biomass suspension: adjusting the pH value 4.0 ~ 6.0 of biomass suspension, temperature dimension
It holds at 45 DEG C ~ 60 DEG C；
Step 5: the preparation and addition of pectase and ferulic acid lipase combination enzyme: proportionally by ferulic acid lipase and pectase
1:3 ~ 1:8 mixed preparing pectase and ferulic acid lipase combine enzyme, be added into biomass suspension 1 ‰ ~ 8 ‰ pectase and
Ferulic acid lipase combination enzyme obtains biomass solution；
Step 6: biomass solution digests: biomass solution digests for 24 hours ~ 60h under the conditions of revolving speed 60r/min ~ 90r/min；
Step 7: adjusting the pH of biomass solution again: adjusting the pH value 4.5 ~ 6.5 of biomass solution；
Step 8: the fine group synthase for tieing up plain enzyme ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase is prepared and addition: will
Xian dimension Su Mei ﹑ dissolubility polysaccharide monooxygenase (proportionally match LPMO) ﹑ catalase by 1:0.2:0.1 ~ 1:0.5:0.7 mixing
The fine three groups of synthase for tieing up plain enzyme ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase of system, are added to biomass solution
0.5 ‰ ~ 1.0% three groups of synthase, add the L-AA of 5mmol/L ~ 10mmol/L；
Step 9: biomass solution digests again: under the conditions of revolving speed 60r/min ~ 90r/min biomass solution digest again 48h ~
90h；
Step 10: separating by filter paper to the solid-liquid in enzymolysis liquid, enzymolysis liquid is obtained, then by enzymolysis liquid in rotary evaporation
It is concentrated in vacuo in instrument, obtains biological syrup.
Further, the biological material is corn stover, corncob, agriculture and forestry organic waste material, sugarcane, wheat, sugar grass
At least one of.
Compared with prior art, the method for the invention for preparing biological syrup using biomass has a characteristic that
(1) form of pectase and ferulic acid lipase combination enzyme can reduce the viscosity of biomass material；
(2) the fine intervention shape for tieing up plain enzyme ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase group synthase and L-AA
Formula not only reduces usage amount and enzymolysis time to cellulase preparation, improves production efficiency and reduces cost；Moreover, may be used also
So that the service life of cellulase preparation extends, reuse ratio is improved；
(3) concentration of syrup in enzymolysis liquid is improved, so that the substrate of high concentration is provided for fermentation, so that subsequent steam consumption subtracts
It is few, reduce cost；
(4) the Fu Chan Wu ﹑ inhibitor generated in enzymolysis liquid is reduced, so that good environment is provided for subsequent fermentation, so that fermentation
Efficiency gets a promotion.
Specific embodiment
In order to which technical problems, technical solutions and advantages to be solved are more clearly understood, tie below
Embodiment is closed, the present invention will be described in further detail.It should be appreciated that specific embodiment described herein is only used to solve
The present invention is released, is not intended to limit the present invention.
The present invention prepares the preferred embodiment of the method for biological syrup using biomass, will using biological material as raw material
The biomass material of certain mesh number is obtained by screening after biological material progress pretreatment early period, biomass material is done
Then dry processing weighs a certain amount of biomass material and prepares certain density suspension, adjust the pH value of suspension, keeps one
Fixed temperature is added a certain amount of pectase and ferulic acid lipase combination enzyme, and is stirred under certain revolving speed, enzymatic hydrolysis one
The section time, then adjust pH value and keep certain temperature, add a certain amount of Xian dimension Su Mei ﹑ dissolubility polysaccharide monooxygenase
(LPMO) the group synthase of ﹑ catalase, the L-AA added digest certain time, measure the Portugal cellulose Zhuan Huashuai ﹑
Then grape sugar concentration is filtered by filter paper, separates to the solid-liquid in enzymolysis liquid, obtain enzymolysis liquid, then enzymolysis liquid exists
It is concentrated in vacuo in Rotary Evaporators, obtains biological syrup.
Be to the principle of biomass cellulase solution saccharification in the methods of the invention: (1) pectin is in plant cell wall
Most complicated polysaccharide, pectase is can to degrade or any polypeptide of modified pectin, by the pectic substance in cell wall
It degrades, obtains small cell substance, such as oligosaccharides or sugar monomer, so that the viscosity of material reduces；(2) ferulic acid
Esterase is to discharge ferulic acid by ferulic acid group-sugar in degradation cell wall and sugar, sugar can be oligosaccharides or polysaccharide, while can
Pectin using in the auxiliary enzymes accelerated decomposition plant cell wall as pectase；(3) Xian ties up Su Mei ﹑ dissolubility polysaccharide monooxygenase
(LPMO) ﹑ catalase group synthase is to be degraded by mutual synergistic effect to cellulose in biomass, and dissolubility is more
Sugared monooxygenase (LPMO) makes the C-H key on cellulose chain be broken abstract hydrogen atoms by oxidation, is inserted into an oxygen original
Son makes glycosidic linkage lose stable state, finally cracks glycosidic bond；The addition of L-AA is added for activation dissolubility polysaccharide list
The activity of oxygenase (LPMO), thus effect of the reinforcing fiber element enzyme to substrate, catalase is to make fibre by oxygen supply means
It ties up plain enzyme enzymolysis speed to accelerate, to accelerate enzymolysis efficiency.
Specifically, the method for preparing biological syrup using biomass of the invention includes the following steps:
Step 1: the pretreatment of biological material: biological material is used steam pre-treatment, dilute acid pretreatment, wet oxidation, use
Wet explosive pretreatment, Biological Pretreatment, the supercritical CO of organic solvent2Pretreatment, overcritical H2O is pre-processed, ozone pre-processes,
Ionic liquid pretreatment, ultrasonic pretreatment, Microwave Pretreatment, γ radiate any one mode in pretreatment and are pre-processed.
Step 2: biological material is dried and screening process: by the dry 48h of pretreated biological material,
Then 40 mesh ~ 60 mesh biomass material is sieved with sieve.
Step 3: preparing the suspension of biomass material: weighing biomass material 40g, be configured to substrate with deionized water
The biomass suspension of concentration 15% ~ 30%.
Step 4: adjusting the pH value and temperature of biomass suspension: adjusting the pH value 4.0 ~ 6.0 of biomass suspension, temperature
Degree maintains 45 DEG C ~ 60 DEG C.
Step 5: the preparation and addition of pectase and ferulic acid lipase combination enzyme: by ferulic acid lipase and pectase according to
Ratio 1:3 ~ 1:8 mixed preparing pectase and ferulic acid lipase combine enzyme, and 1 ‰ ~ 8 ‰ pectin is added into biomass suspension
Enzyme and ferulic acid lipase combination enzyme obtain biomass solution.
Step 6: biomass solution digest: under the conditions of revolving speed 60r/min ~ 90r/min biomass solution enzymatic hydrolysis for 24 hours ~
60h。
Step 7: adjusting the pH of biomass solution again: adjusting the pH value 4.5 ~ 6.5 of biomass solution.
Step 8: the fine group synthase for tieing up plain enzyme ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase is prepared and is added
Add: Xian is tieed up into Su Mei ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase proportionally 1:0.2:0.1 ~ 1:0.5:0.7
Mixed preparing fibre ties up three groups of synthase of plain enzyme ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase, to biomass solution plus
Three groups of synthase for entering 0.5 ‰ ~ 1.0%, add the L-AA of 5mmol/L ~ 10mmol/L.
Step 9: biomass solution digests again: biomass solution digests again under the conditions of revolving speed 60r/min ~ 90r/min
48h~90h。
Step 10: separating by filter paper to the solid-liquid in enzymolysis liquid, enzymolysis liquid is obtained, is then rotating enzymolysis liquid
It is concentrated in vacuo in evaporimeter, obtains biological syrup.
The biological material is corn stover, corncob, agriculture and forestry organic waste material, sugarcane, wheat, at least one in sugar grass
Kind.The cellulase includes that two carbohydrase ﹑ β-glucosyl enzym of cellobiose water solution enzyme ﹑ fiber is cut outside endoglucanase (EG) ﹑
(BG)。
After measured, after using method processing of the invention, the concentration of glucose in biomass solution reach 80g/l ~
125.5g/l, cellulose conversion ratio reach 85% ~ 96.5%.
Further illustrate the method for the invention for preparing biological syrup using biomass combined with specific embodiments below.
Embodiment 1.
The first described method for preparing biological syrup using biomass, includes the following steps:
The pretreatment of A1, biological material: cornstalk biological material is pre-processed using steam pre-treatment mode.
A2, biological material is dried and screening process: pretreated cornstalk biological material is dry
Then 48h sieves 40 mesh ~ 60 mesh cornstalk biological matter material with sieve.
A3, the suspension for preparing biomass material: cornstalk biological matter material 40g is weighed, is configured to deionized water
The cornstalk biological matter suspension of concentration of substrate 15%.
A4, the pH value and temperature for adjusting biomass suspension: the pH value 4.0 of adjusting corn straw biomass suspension, temperature
Degree maintains 45 DEG C.
The preparation and addition of A5, pectase and ferulic acid lipase combination enzyme: proportionally by ferulic acid lipase and pectase
1:3 mixed preparing pectase and ferulic acid lipase combine enzyme, be added into cornstalk biological matter suspension 1 ‰ pectase and
Ferulic acid lipase combination enzyme obtains cornstalk biological matter solution.
A6, biomass solution enzymatic hydrolysis: cornstalk biological matter in-solution digestion 48h under the conditions of revolving speed 60r/min.
A7, the pH for adjusting biomass solution again: the pH value 4.5 of adjusting corn straw biomass solution.
A8, the fine group synthase for tieing up plain enzyme ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase is prepared and addition: will
Xian ties up Su Mei ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase proportionally 1:0.2:0.1 mixed preparing cellulose
1.0% three groups of synthase are added to cornstalk biological matter solution, add for three groups of synthase of enzyme ﹑ GH61 enzyme ﹑ catalase
The L-AA of 5mmol/L.
A9, biomass solution digest again: cornstalk biological matter solution digests 72h again under the conditions of revolving speed 60r/min.
A10, the solid-liquid in enzymolysis liquid is separated by filter paper, obtains enzymolysis liquid, then steam enzymolysis liquid in rotation
It is concentrated in vacuo in hair instrument, obtains biological syrup.
Embodiment 2.
Second of method for preparing biological syrup using biomass, includes the following steps:
The pretreatment of B1, biological material: corncob biological material is pre-processed using dilute acid pretreatment mode.
B2, biological material is dried and screening process: pretreated corncob biological material is dry
Then 48h sieves 40 mesh ~ 60 mesh corncob biomass material with sieve.
B3, the suspension for preparing biomass material: corncob biomass material 40g is weighed, is configured to bottom with deionized water
The corncob biomass suspension of object concentration 15%.
B4, the pH value and temperature for adjusting biomass suspension: the pH value 6.0 of adjusting corn core biomass suspension, temperature
Maintain 60 DEG C.
The preparation and addition of B5, pectase and ferulic acid lipase combination enzyme: proportionally by ferulic acid lipase and pectase:
153 mixed preparing pectases and ferulic acid lipase combine enzyme, and 1 ‰ pectase and asafoetide are added into corncob biomass suspension
Sour lipase group synthase obtains biomass solution.
B6, biomass solution enzymatic hydrolysis: corncob biomass solution digests 60h under the conditions of revolving speed 60r/min.
B7, the pH for adjusting biomass solution again: the pH value 4.5 of adjusting corn core biomass solution.
B8, the fine group synthase for tieing up plain enzyme ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase is prepared and addition: will
Xian ties up Su Mei ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase proportionally 1:0.5:0.7 mixed preparing cellulose
Three groups of synthase of enzyme ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase are added 1.0% to corncob biomass solution
Three groups of synthase, add 5mmol/L- ascorbic acid.
B9, biomass solution digest again: corncob biomass solution digests 72h again under the conditions of revolving speed 60r/min.
B10, the solid-liquid in enzymolysis liquid is separated by filter paper, obtains enzymolysis liquid, then steam enzymolysis liquid in rotation
It is concentrated in vacuo in hair instrument, obtains biological syrup.
Embodiment 3.
The third described method for preparing biological syrup using biomass, includes the following steps:
The pretreatment of C1, biological material: sugarcane biological material is pre-processed using wet oxidation mode.
C2, biological material is dried and screening process: by the dry 48h of pretreated sugarcane biological material,
Then 40 mesh ~ 60 mesh sugarcane biomass material is sieved with sieve.
C3, the suspension for preparing biomass material: biomass material 40g is weighed, is configured to concentration of substrate with deionized water
30% biomass suspension.
C4, the pH value and temperature for adjusting biomass suspension: the pH value 5.5 of biomass suspension is adjusted, temperature maintains
55℃。
The preparation and addition of C5, pectase and ferulic acid lipase combination enzyme: proportionally by ferulic acid lipase and pectase:
1:5 mixed preparing pectase and ferulic acid lipase combine enzyme, and 8 ‰ pectase and orizanol are added into biomass suspension
Enzyme group synthase obtains biomass solution.
C6, biomass solution enzymatic hydrolysis: biomass solution digests for 24 hours under the conditions of revolving speed 90r/min.
C7, the pH for adjusting biomass solution again: the pH value 4.5 of biomass solution is adjusted.
C8, the fine group synthase for tieing up plain enzyme ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase is prepared and addition: will
Xian ties up Su Mei ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase proportionally 1:0.3:0.5 mixed preparing cellulose
0.6% three groups of synthase are added to biomass solution, add 7mmol/L's for three groups of synthase of enzyme ﹑ GH61 enzyme ﹑ catalase
L-AA.
C9, biomass solution digest again: biomass solution digests 90h again under the conditions of revolving speed 90r/min.
C10, the solid-liquid in enzymolysis liquid is separated by filter paper, obtains enzymolysis liquid, then steam enzymolysis liquid in rotation
It is concentrated in vacuo in hair instrument, obtains biological syrup.
Embodiment 4.
The 4th kind of method for preparing biological syrup using biomass, includes the following steps:
The pretreatment of D1, biological material: wheat biological material is used into overcritical H2O pretreatment mode is pre-processed.
D2, biological material is dried and screening process: by the dry 48h of pretreated wheat biological material,
Then 40 mesh ~ 60 mesh wheat biomass material is sieved with sieve.
D3, the suspension for preparing biomass material: wheat biomass material 40g is weighed, is configured to substrate with deionized water
The wheat biomass suspension of concentration 30%.
D4, the pH value and temperature for adjusting biomass suspension: the pH value 5.0 of wheat biomass suspension, temperature dimension are adjusted
It holds at 50 DEG C.
The preparation and addition of D5, pectase and ferulic acid lipase combination enzyme: proportionally by ferulic acid lipase and pectase:
1:3 mixed preparing pectase and ferulic acid lipase combine enzyme, and 5 ‰ pectase and asafoetide are added into wheat biomass suspension
Sour lipase group synthase obtains wheat biomass solution.
D6, biomass solution enzymatic hydrolysis: wheat biomass solution digests 48h under the conditions of revolving speed 80r/min.
D7, the pH for adjusting biomass solution again: the pH value 6.5 of wheat biomass solution is adjusted.
D8, the fine group synthase for tieing up plain enzyme ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase is prepared and addition: will
Xian ties up Su Mei ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase proportionally 1:0.5:0.5 mixed preparing cellulose
Three groups of synthase of enzyme ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase are added the three of 0.8% to wheat biomass solution
Group synthase, adds the L-AA of 8mmol/L.
D9, biomass solution digest again: wheat biomass solution digests 80h again under the conditions of revolving speed 80r/min.
D10, the solid-liquid in enzymolysis liquid is separated by filter paper, obtains enzymolysis liquid, then steam enzymolysis liquid in rotation
It is concentrated in vacuo in hair instrument, obtains biological syrup.
Embodiment 5.
The 5th kind of method for preparing biological syrup using biomass, includes the following steps:
The pretreatment of E1, biological material: sugar grass biological material is used into overcritical H2O pretreatment mode is located in advance
Reason.
E2, biological material is dried and screening process: pretreated sugar grass biological material is dry
Then 48h sieves 40 mesh ~ 60 mesh sugar grass biomass material with sieve.
E3, the suspension for preparing biomass material: sugar grass biomass material 40g is weighed, is configured to bottom with deionized water
The sugar grass biomass suspension of object concentration 15%.
E4, the pH value and temperature for adjusting biomass suspension: the pH value 4.0 of sugar grass biomass suspension, temperature are adjusted
Maintain 45 DEG C.
The preparation and addition of E5, pectase and ferulic acid lipase combination enzyme: proportionally by ferulic acid lipase and pectase
1:5 mixed preparing pectase and ferulic acid lipase combine enzyme, be added into sugar grass biomass suspension 8 ‰ pectase and Ah
Wei's acid lipase group synthase obtains sugar grass biomass solution.
E6, biomass solution enzymatic hydrolysis: sugar grass biomass solution digests 48h under the conditions of revolving speed 60r/min.
E7, the pH for adjusting biomass solution again: the pH value 4.5 of sugar grass biomass solution is adjusted.
E8, the fine group synthase for tieing up plain enzyme ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase is prepared and addition: will
Xian ties up Su Mei ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase proportionally 1:0.5:0.5 mixed preparing cellulose
Three groups of synthase of enzyme ﹑ dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase are added 0.5% to sugar grass biomass solution
Three groups of synthase, add the L-AA of 10mmol/L.
E9, biomass solution digest again: sugar grass biomass solution digests 80h again under the conditions of revolving speed 80r/min.
E10, the solid-liquid in enzymolysis liquid is separated by filter paper, obtains enzymolysis liquid, then steam enzymolysis liquid in rotation
It is concentrated in vacuo in hair instrument, obtains biological syrup.
After measured, in embodiment 1, after cornstalk biological material is using method processing of the invention, biomass
Concentration of glucose in solution reaches 80g/L, and cellulose conversion ratio reaches 85%.In example 2, corncob biological material
After method processing of the invention, the concentration of glucose in biomass solution reaches 125.5g/L, and cellulose conversion ratio reaches
To 96.5%.In embodiment 3, after sugarcane biological material is using method processing of the invention, the grape in biomass solution
Sugared concentration reaches 98.8g/L, and cellulose conversion ratio reaches 88%.In example 4, wheat biological material is using of the invention
After method processing, the concentration of glucose in biomass solution reaches 110.4g/L, and cellulose conversion ratio reaches 92.8%.In reality
It applies in example 5, after sugar grass biological material is using method processing of the invention, the concentration of glucose in biomass solution reaches
92.4g/L, cellulose conversion ratio reach 87.9%.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (8)
1. a kind of method for preparing biological syrup using biomass, which is characterized in that using biological material as raw material, by biomass
The biomass material of certain mesh number is obtained by screening after material progress pretreatment early period, place is dried in biomass material
Then reason weighs a certain amount of biomass material and prepares certain density suspension, adjusts the pH value of suspension, keep certain
Temperature is added a certain amount of pectase and ferulic acid lipase combination enzyme, and is stirred under certain revolving speed, when digesting one section
Between, then adjust pH value and keep certain temperature, it adds a certain amount of Xian dimension Su Mei ﹑ dissolubility polysaccharide Dan and adds Yang Mei ﹑ peroxide
The group synthase for changing hydrogen enzyme adds a certain amount of L-AA, digests certain time, is then filtered by filter paper, to enzymatic hydrolysis
Solid-liquid in liquid is separated, and enzymolysis liquid is obtained, and is then concentrated in vacuo enzymolysis liquid in Rotary Evaporators, is obtained biology
Syrup.
2. the method for preparing biological syrup using biomass as described in claim 1, which is characterized in that described to utilize biomass
The method for preparing biological syrup includes the following steps:
Step 1: the pretreatment of biological material: biological material is used steam pre-treatment, dilute acid pretreatment, wet oxidation, use
Wet explosive pretreatment, Biological Pretreatment, the supercritical CO of organic solvent2Pretreatment, overcritical H2O is pre-processed, ozone pre-processes,
Ionic liquid pretreatment, ultrasonic pretreatment, Microwave Pretreatment, γ radiate any one mode in pretreatment and are pre-processed；
Step 2: biological material is dried and screening process: by the dry 48h of pretreated biological material, then
40 mesh ~ 60 mesh biomass material is sieved with sieve；
Step 3: preparing the suspension of biomass material: weighing biomass material 40g, be configured to concentration of substrate with deionized water
15% ~ 30% biomass suspension；
Step 4: adjusting the pH value and temperature of biomass suspension: adjusting the pH value 4.0 ~ 6.0 of biomass suspension, temperature dimension
It holds at 45 DEG C ~ 60 DEG C；
Step 5: the preparation and addition of pectase and ferulic acid lipase combination enzyme: proportionally by ferulic acid lipase and pectase
1:3 ~ 1:8 mixed preparing pectase and ferulic acid lipase combine enzyme, be added into biomass suspension 1 ‰ ~ 8 ‰ pectase and
Ferulic acid lipase combination enzyme obtains biomass solution；
Step 6: biomass solution digests: biomass solution digests for 24 hours ~ 60h under the conditions of revolving speed 60r/min ~ 90r/min；
Step 7: adjusting the pH of biomass solution again: adjusting the pH value 4.5 ~ 6.5 of biomass solution；
Step 8: the fine preparation of group synthase and addition tieed up plain enzyme ﹑ dissolubility polysaccharide list and add oxygen enzyme ﹑ catalase: by cellulose
Mei ﹑ dissolubility polysaccharide Dan adds Yang Mei ﹑ catalase, and proportionally 1:0.2:0.1 ~ 1:0.5:0.7 mixed preparing Xian ties up Su Mei ﹑
Three groups of synthase of dissolubility polysaccharide monooxygenase (LPMO) ﹑ catalase are added the three of 0.5 ‰ ~ 1.0% to biomass solution
Group synthase, is added the L-AA of 5mmol/L ~ 10mmol/L；
Step 9: biomass solution digests again: under the conditions of revolving speed 60r/min ~ 90r/min biomass solution digest again 48h ~
90h；
Step 10: separating by filter paper to the solid-liquid in enzymolysis liquid, enzymolysis liquid is obtained, then by enzymolysis liquid in rotary evaporation
It is concentrated in vacuo in instrument, obtains biological syrup.
3. the method for preparing biological syrup using biomass as claimed in claim 2, which is characterized in that the biological material
For at least one of corn stover, corncob, agriculture and forestry organic waste material, sugarcane, wheat, sugar grass.
4. the method for preparing biological syrup using biomass as claimed in claim 2 or claim 3, which is characterized in that described to utilize life
The method that substance prepares biological syrup includes the following steps:
The pretreatment of A1, biological material: cornstalk biological material is pre-processed using steam pre-treatment mode；
A2, biological material is dried and screening process: by the dry 48h of pretreated cornstalk biological material,
Then 40 mesh ~ 60 mesh cornstalk biological matter material is sieved with sieve；
A3, the suspension for preparing biomass material: cornstalk biological matter material 40g is weighed, is configured to substrate with deionized water
The cornstalk biological matter suspension of concentration 15%；
A4, the pH value and temperature for adjusting biomass suspension: the pH value 4.0 of adjusting corn straw biomass suspension, temperature dimension
It holds at 45 DEG C；
The preparation and addition of A5, pectase and ferulic acid lipase combination enzyme: by ferulic acid lipase and pectase, proportionally 1:3 is mixed
It closes and prepares pectase and ferulic acid lipase combination enzyme, 1 ‰ pectase and asafoetide are added into cornstalk biological matter suspension
Sour lipase group synthase obtains cornstalk biological matter solution；
A6, biomass solution enzymatic hydrolysis: cornstalk biological matter in-solution digestion 48h under the conditions of revolving speed 60r/min；
A7, the pH for adjusting biomass solution again: the pH value 4.5 of adjusting corn straw biomass solution；
A8, the fine preparation of group synthase and addition tieed up plain enzyme ﹑ dissolubility polysaccharide list and add oxygen enzyme ﹑ catalase: it is molten that fibre is tieed up into plain enzyme ﹑
Solution property polysaccharide Dan adds Yang Mei ﹑ catalase, and proportionally 1:0.2:0.1 mixed preparing Xian ties up Su Mei ﹑ dissolubility polysaccharide list oxygenation
1.0% three groups of synthase are added to cornstalk biological matter solution, add 5mmol/L for three groups of synthase of enzyme ﹑ catalase
L-AA；
A9, biomass solution digest again: cornstalk biological matter solution digests 72h again under the conditions of revolving speed 60r/min；
A10, the solid-liquid in enzymolysis liquid is separated by filter paper, enzymolysis liquid is obtained, then by enzymolysis liquid in Rotary Evaporators
In be concentrated in vacuo, obtain biological syrup.
5. the method for preparing biological syrup using biomass as claimed in claim 2 or claim 3, which is characterized in that described to utilize life
The method that substance prepares biological syrup includes the following steps:
The pretreatment of B1, biological material: corncob biological material is pre-processed using dilute acid pretreatment mode；
B2, biological material is dried and screening process: by the dry 48h of pretreated corncob biological material, so
40 mesh ~ 60 mesh corncob biomass material is sieved with sieve afterwards；
B3, the suspension for preparing biomass material: corncob biomass material 40g is weighed, it is dense to be configured to substrate with deionized water
The corncob biomass suspension of degree 15%；
B4, the pH value and temperature for adjusting biomass suspension: the pH value 6.0 of adjusting corn core biomass suspension, temperature maintain
At 60 DEG C；
The preparation and addition of B5, pectase and ferulic acid lipase combination enzyme: proportionally by ferulic acid lipase and pectase: 153
Mixed preparing pectase and ferulic acid lipase combine enzyme, and 1 ‰ pectase and ferulic acid are added into corncob biomass suspension
Lipase group synthase obtains biomass solution；
B6, biomass solution enzymatic hydrolysis: corncob biomass solution digests 60h under the conditions of revolving speed 60r/min；
B7, the pH for adjusting biomass solution again: the pH value 4.5 of adjusting corn core biomass solution；
B8, the fine preparation of group synthase and addition tieed up plain enzyme ﹑ dissolubility polysaccharide list and add oxygen enzyme ﹑ catalase: it is molten that fibre is tieed up into plain enzyme ﹑
Solution property polysaccharide Dan adds Yang Mei ﹑ catalase, and proportionally 1:0.5:0.7 mixed preparing Xian ties up Su Mei ﹑ dissolubility polysaccharide list oxygenation
1.0% three groups of synthase are added to corncob biomass solution, add 5mmol/L's for three groups of synthase of enzyme ﹑ catalase
L-AA；
B9, biomass solution digest again: corncob biomass solution digests 72h again under the conditions of revolving speed 60r/min；
B10, the solid-liquid in enzymolysis liquid is separated by filter paper, enzymolysis liquid is obtained, then by enzymolysis liquid in Rotary Evaporators
In be concentrated in vacuo, obtain biological syrup.
6. the method for preparing biological syrup using biomass as claimed in claim 2 or claim 3, which is characterized in that described to utilize life
The method that substance prepares biological syrup includes the following steps:
The pretreatment of C1, biological material: sugarcane biological material is pre-processed using wet oxidation mode；
C2, biological material is dried and screening process: by the dry 48h of pretreated sugarcane biological material, then
40 mesh ~ 60 mesh sugarcane biomass material is sieved with sieve；
C3, the suspension for preparing biomass material: biomass material 40g is weighed, is configured to concentration of substrate 30% with deionized water
Biomass suspension；
C4, the pH value and temperature for adjusting biomass suspension: the pH value 5.5 of biomass suspension is adjusted, temperature maintains 55 DEG C；
The preparation and addition of C5, pectase and ferulic acid lipase combination enzyme: proportionally by ferulic acid lipase and pectase: 1:5
Mixed preparing pectase and ferulic acid lipase combine enzyme, and 8 ‰ pectase and ferulic acid lipase are added into biomass suspension
Group synthase obtains biomass solution；
C6, biomass solution enzymatic hydrolysis: biomass solution digests for 24 hours under the conditions of revolving speed 90r/min；
C7, the pH for adjusting biomass solution again: the pH value 4.5 of biomass solution is adjusted；
C8, the fine preparation of group synthase and addition tieed up plain enzyme ﹑ dissolubility polysaccharide list and add oxygen enzyme ﹑ catalase: it is molten that fibre is tieed up into plain enzyme ﹑
Solution property polysaccharide Dan adds Yang Mei ﹑ catalase, and proportionally 1:0.3:0.5 mixed preparing Xian ties up Su Mei ﹑ dissolubility polysaccharide list oxygenation
Three groups of synthase of enzyme ﹑ catalase, 0.6% three groups of synthase are added to biomass solution, and the L- for adding 7mmol/L is anti-bad
Hematic acid；
C9, biomass solution digest again: biomass solution digests 90h again under the conditions of revolving speed 90r/min；
C10, the solid-liquid in enzymolysis liquid is separated by filter paper, enzymolysis liquid is obtained, then by enzymolysis liquid in Rotary Evaporators
In be concentrated in vacuo, obtain biological syrup.
7. the method for preparing biological syrup using biomass as claimed in claim 2 or claim 3, which is characterized in that described to utilize life
The method that substance prepares biological syrup includes the following steps:
The pretreatment of D1, biological material: wheat biological material is used into overcritical H2O pretreatment mode is pre-processed；
D2, biological material is dried and screening process: by the dry 48h of pretreated wheat biological material, then
40 mesh ~ 60 mesh wheat biomass material is sieved with sieve；
D3, the suspension for preparing biomass material: wheat biomass material 40g is weighed, is configured to concentration of substrate with deionized water
30% wheat biomass suspension；
D4, the pH value and temperature for adjusting biomass suspension: the pH value 5.0 of wheat biomass suspension is adjusted, temperature maintains
50℃；
The preparation and addition of D5, pectase and ferulic acid lipase combination enzyme: proportionally by ferulic acid lipase and pectase: 1:3
Mixed preparing pectase and ferulic acid lipase combine enzyme, and 5 ‰ pectase and ferulic acid are added into wheat biomass suspension
Lipase group synthase obtains wheat biomass solution；
D6, biomass solution enzymatic hydrolysis: wheat biomass solution digests 48h under the conditions of revolving speed 80r/min；
D7, the pH for adjusting biomass solution again: the pH value 6.5 of wheat biomass solution is adjusted；
D8, the fine preparation of group synthase and addition tieed up plain enzyme ﹑ dissolubility polysaccharide list and add oxygen enzyme ﹑ catalase: it is molten that fibre is tieed up into plain enzyme ﹑
Solution property polysaccharide Dan adds Yang Mei ﹑ catalase, and proportionally 1:0.5:0.5 mixed preparing Xian ties up Su Mei ﹑ dissolubility polysaccharide list oxygenation
0.8% three groups of synthase are added to wheat biomass solution, add the L- of 8mmol/L for three groups of synthase of enzyme ﹑ catalase
Ascorbic acid；
D9, biomass solution digest again: wheat biomass solution digests 80h again under the conditions of revolving speed 80r/min；
D10, the solid-liquid in enzymolysis liquid is separated by filter paper, enzymolysis liquid is obtained, then by enzymolysis liquid in Rotary Evaporators
In be concentrated in vacuo, obtain biological syrup.
8. the method for preparing biological syrup using biomass as claimed in claim 2 or claim 3, which is characterized in that described to utilize life
The method that substance prepares biological syrup includes the following steps:
The pretreatment of E1, biological material: sugar grass biological material is used into overcritical H2O pretreatment mode is pre-processed；
E2, biological material is dried and screening process: by the dry 48h of pretreated sugar grass biological material, so
40 mesh ~ 60 mesh sugar grass biomass material is sieved with sieve afterwards；
E3, the suspension for preparing biomass material: sugar grass biomass material 40g is weighed, it is dense to be configured to substrate with deionized water
The sugar grass biomass suspension of degree 15%；
E4, the pH value and temperature for adjusting biomass suspension: adjusting the pH value 4.0 of sugar grass biomass suspension, and temperature maintains
At 45 DEG C；
The preparation and addition of E5, pectase and ferulic acid lipase combination enzyme: by ferulic acid lipase and pectase, proportionally 1:5 is mixed
It closes and prepares pectase and ferulic acid lipase combination enzyme, 8 ‰ pectase and ferulic acid are added into sugar grass biomass suspension
Lipase group synthase obtains sugar grass biomass solution；
E6, biomass solution enzymatic hydrolysis: sugar grass biomass solution digests 48h under the conditions of revolving speed 60r/min；
E7, the pH for adjusting biomass solution again: the pH value 4.5 of sugar grass biomass solution is adjusted；
E8, the fine preparation of group synthase and addition tieed up plain enzyme ﹑ dissolubility polysaccharide list and add oxygen enzyme ﹑ catalase: it is molten that fibre is tieed up into plain enzyme ﹑
Solution property polysaccharide Dan adds Yang Mei ﹑ catalase, and proportionally 1:0.5:0.5 mixed preparing Xian ties up Su Mei ﹑ dissolubility polysaccharide list oxygenation
0.5% three groups of synthase are added to sugar grass biomass solution, add 10mmol/L's for three groups of synthase of enzyme ﹑ catalase
L-AA；
E9, biomass solution digest again: sugar grass biomass solution digests 80h again under the conditions of revolving speed 80r/min；
E10, the solid-liquid in enzymolysis liquid is separated by filter paper, enzymolysis liquid is obtained, then by enzymolysis liquid in Rotary Evaporators
In be concentrated in vacuo, obtain biological syrup.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103703139A (en) * | 2011-07-22 | 2014-04-02 | 诺维信北美公司 | Processes for pretreating cellulosic material and improving hydrolysis thereof |
| CN105567761A (en) * | 2014-10-16 | 2016-05-11 | 中粮营养健康研究院有限公司 | Enzymolysis method for cellulose-contained material |
| CN106460020A (en) * | 2014-04-30 | 2017-02-22 | 帝斯曼知识产权资产管理有限公司 | Process for enzymatic hydrolysis of lignocellulosic material and fermentation of sugars |
| WO2017097897A1 (en) * | 2015-12-10 | 2017-06-15 | Dsm Ip Assets B.V. | Beta-glucosidase and uses thereof |
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2018
- 2018-11-22 CN CN201811402058.3A patent/CN109371077A/en not_active Withdrawn
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103703139A (en) * | 2011-07-22 | 2014-04-02 | 诺维信北美公司 | Processes for pretreating cellulosic material and improving hydrolysis thereof |
| CN106460020A (en) * | 2014-04-30 | 2017-02-22 | 帝斯曼知识产权资产管理有限公司 | Process for enzymatic hydrolysis of lignocellulosic material and fermentation of sugars |
| CN105567761A (en) * | 2014-10-16 | 2016-05-11 | 中粮营养健康研究院有限公司 | Enzymolysis method for cellulose-contained material |
| WO2017097897A1 (en) * | 2015-12-10 | 2017-06-15 | Dsm Ip Assets B.V. | Beta-glucosidase and uses thereof |
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| BRIAN R等: ""Catalase improves saccharification of lignocellulose by reducing lytic polysaccharide monooxygenase-associated enzyme inactivation"", 《BIOTECHNOL LETT》 * |
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