CN117051055B - Method for preparing low inhibitor straw sugar suitable for fermentation and catalysis by using uniform graded straw - Google Patents
Method for preparing low inhibitor straw sugar suitable for fermentation and catalysis by using uniform graded straw Download PDFInfo
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- CN117051055B CN117051055B CN202311317653.8A CN202311317653A CN117051055B CN 117051055 B CN117051055 B CN 117051055B CN 202311317653 A CN202311317653 A CN 202311317653A CN 117051055 B CN117051055 B CN 117051055B
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- 239000010902 straw Substances 0.000 title claims abstract description 354
- 238000000034 method Methods 0.000 title claims abstract description 101
- 235000000346 sugar Nutrition 0.000 title claims abstract description 79
- 239000003112 inhibitor Substances 0.000 title claims abstract description 76
- 238000000855 fermentation Methods 0.000 title claims abstract description 32
- 230000004151 fermentation Effects 0.000 title claims abstract description 32
- 238000006555 catalytic reaction Methods 0.000 title claims abstract description 24
- 238000004880 explosion Methods 0.000 claims abstract description 88
- 238000011282 treatment Methods 0.000 claims abstract description 73
- 238000005096 rolling process Methods 0.000 claims abstract description 67
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- 238000012216 screening Methods 0.000 claims abstract description 17
- 240000008042 Zea mays Species 0.000 claims description 89
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- 239000000243 solution Substances 0.000 claims description 46
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- XPFJYKARVSSRHE-UHFFFAOYSA-K trisodium;2-hydroxypropane-1,2,3-tricarboxylate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].[Na+].OC(=O)CC(O)(C(O)=O)CC(O)=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O XPFJYKARVSSRHE-UHFFFAOYSA-K 0.000 claims description 13
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- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 claims description 2
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- VQHSOMBJVWLPSR-WUJBLJFYSA-N maltitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H]([C@H](O)CO)O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O VQHSOMBJVWLPSR-WUJBLJFYSA-N 0.000 claims description 2
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- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 4
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- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 2
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- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 description 1
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 description 1
- ISPYRSDWRDQNSW-UHFFFAOYSA-L manganese(II) sulfate monohydrate Chemical compound O.[Mn+2].[O-]S([O-])(=O)=O ISPYRSDWRDQNSW-UHFFFAOYSA-L 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
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- 150000002772 monosaccharides Chemical class 0.000 description 1
- 239000012452 mother liquor Substances 0.000 description 1
- 229910052759 nickel Inorganic materials 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
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Classifications
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P2201/00—Pretreatment of cellulosic or lignocellulosic material for subsequent enzymatic treatment or hydrolysis
-
- 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
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
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- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention provides a method for preparing low inhibitor straw sugar suitable for fermentation and catalysis by homogenizing graded straw, which comprises the following steps: carrying out rolling treatment on the refined straw to obtain rolled straw; setting mechanical-chemical treatment parameters according to a mechanical grading torsion mechanical model, and carrying out mechanical grading treatment on the rolled straw by using a mechanical-chemical treatment method to obtain mechanical graded straw; screening the mechanically graded straw to obtain graded straw; carrying out rehydration treatment and steam explosion treatment on the graded straw to obtain a low inhibitor straw sugar production raw material; and carrying out high-solid enzymolysis treatment on the low-inhibitor straw raw material to obtain the low-inhibitor straw sugar. The method is based on the mechanical property difference of the straw raw materials, based on a straw homogenization grading model and a steam explosion process inhibitor generation model, adopts grading equipment with simple operation, has high raw material homogenization degree, good inhibitor reduction effect and strong theoretical basis, and has strong industrialized applicability.
Description
Technical Field
The invention belongs to the field of biomass utilization, relates to a method for preparing low inhibitor straw sugar, and in particular relates to a method for preparing low inhibitor straw sugar suitable for fermentation and catalysis by homogenizing and grading straw.
Background
In the development of the modern society, biomass resources are taken as the most abundant renewable resources, and can be considered to lead the substitution of the modern manufacturing industries such as biological fermentation, chemical catalysis and the like to the traditional petrochemical industry, thereby promoting the industrial development of the new era. The utilization of biomass resources in modern industry requires the conversion of biomass resources into available glucose, xylose and other sugar resources by biorefinery. However, the presence of organic substances such as small-molecule organic acids, furan compounds, and phenolic acid compounds, which are produced during the biomass refining process, in the sugar solution can inhibit the growth of most of the cells, and can also cause the poisoning and deactivation of most of the catalysts. This also results in the fact that modern industrial technicians are constantly performing cell engineering and catalyst modification to adapt the cells and catalyst to high inhibitor concentrations of sugar solutions; or physical adsorption, chemical detoxification and the like are carried out to remove the inhibitor in the lignocellulose sugar solution, thereby increasing the availability of the lignocellulose sugar solution. The lignocellulose sugar solution detoxification process greatly increases the use cost of the lignocellulose sugar solution, so that the biological manufacturing industry and the chemical catalysis industry have no good renewable resource supply, and the development of biomass resource utilization and modern manufacturing industry is greatly hindered.
Research shows that most of the inhibitors of lignocellulose sugar solution originate from the lignocellulose pretreatment stage, and because biomass resources have compact degradation resistance barriers, strong physicochemical pretreatment is generally adopted in the biomass refining pretreatment stage, so that the degradation resistance barriers of lignocellulose are fully destroyed, and more enzymolysis contact sites are provided for the subsequent enzymolysis process. The strong physicochemical pretreatment can lead to the generation of inhibitory substances such as organic acids, furan compounds and the like with small molecules by excessively pretreating the easily decomposed parts in lignocellulose. While pretreatment intensity is positively correlated with inhibitor concentration generation. In order to meet the carbon source concentration required by bacteria in the biological manufacturing industry and reduce the production cost of lignocellulose fermentable sugar, the pretreatment of lignocellulose generally adopts a high-solid process to strengthen enzymolysis measures, so that high-concentration fermentable sugar is rapidly prepared, and in the production process, the inhibitor concentration can be up to about 2-4%, which also leads to stronger inhibition effect of the produced high-concentration sugar solution on bacteria and catalysts. The generation of the inhibitor needs to be reduced, the pretreatment strength of biological refining and the solid concentration in the enzymolysis process are reduced, the production of lignocellulose sugar solution is greatly reduced, and the production cost of the sugar solution is increased. Therefore, inhibitors become the biggest obstacle to the use of lignocellulose sugar solution.
Meanwhile, the existing inhibitor removal is based on the sugar solution, and the production and processing cost of the sugar solution is greatly increased by aiming at the technologies of physical adsorption, chemical treatment, biological treatment and the like of the sugar solution, and the existing inhibitor removal technology is not well matched with the sugar solution, and after various inhibitor removal technologies are combined for use, an efficient inhibitor removal method capable of realizing industrial production is not provided. The invention starts from the source, establishes a steam explosion process energy model and an inhibitor generation model by analyzing energy distribution in the steam explosion pretreatment process, takes the model as a guiding basis, analyzes the relation between the inhibitor generation and pretreatment process parameters and raw material characteristics, and discovers that the elastic modulus and poisson ratio of the raw material play an important role in the inhibitor generation in the pretreatment process.
Straw, which is the most representative lignocellulose, is mainly composed of cellulose, hemicellulose and lignin. From the analysis on organs, the straw consists of skin, pulp, leaves and the like, wherein the skin-pulp part consists of parenchyma and vascular tissues, the parenchyma has smaller elastic modulus, lower mechanical strength and the vascular tissues have larger elastic modulus and higher mechanical strength, the vascular tissues contained in the straw skin part are about 70 percent, and the pulp part contains about 70 percent; the structural difference causes great difference of mechanical strength of the skin and pulp parts of the straw and the whole straw is not uniform. In the prior art, the processing utilization of all components of the straw is considered, so that the degradation resistance barrier of the whole straw is damaged, the enzymolysis performance of the pretreated straw is improved, the pretreatment strength is too high, and the components with lower elastic modulus are damaged into oligosaccharide or monosaccharide components in the pretreatment process, so that the components are further converted into organic substances such as small molecular organic acid, furan compounds, phenolic acid compounds and the like in the high-strength physicochemical pretreatment, and the growth metabolism of thalli in the fermentation process is greatly and obviously inhibited, thereby reducing the production efficiency and the production benefit of the straw sugar solution.
Because of the heterogeneity of the straw, the pretreatment intensity in the biological refining process is high, the inhibitor is generated more, and the generation of the inhibitor is closely related to the uniformity of the raw materials. However, the existing researches and techniques do not fully research and recognize the characteristics of the raw materials aiming at the heterogeneity of the straw, and some technicians adopt various physicochemical methods or mechanical treatment methods to relatively homogenize the raw materials, but no grading method and grading theory based on the characteristics of the straw raw materials exist.
Aiming at the heterogeneity of the straw, the existing grading technology mainly aims at grading utilization of chemical composition of the corn straw, and adopts an acid treatment and alkali treatment mode to separate cellulose, hemicellulose and lignin. But the purity of the separated products is low, the pollution in the separation process is large, the usability of the separated products is poor, and a small number of patents are from the mechanical separation point of view, but the characteristics of the raw materials are not fully known, only the mechanical is used for crushing and separating the straws, the physical property difference and theoretical guidance of the straws are not highlighted, and the inhibitor for reducing the straw enzymolysis sugar solution from the source is not realized.
CN 102490238A discloses a method for separating the skin and pulp of corn stalks, which uses the existing separator to crush the corn stalks, then performs air separation after sieving, blows away the part with lighter density, and leaves the part with greater density for producing the corn stalk skin particle board. The crushing mode of the method is mainly applied to the process of producing the particle board by using the corn straw, loose pulp parts are removed, the use strength of the skin particle board is improved, and further conversion and utilization of the separated pulp parts are not considered.
CN 115889396a discloses a field crop straw grading method and device based on industrial utilization, which comprises mechanically fluffing straw and grading the straw in a centrifugal shaft type, separating the vascular tissue and the thin-wall tissue of the straw, and obtaining a skin material and a graded straw material. The method mainly starts from a straw field collection and classification device, adopts a plurality of grinding rolls to carry out mechanical fluffing, focuses on the design of a field rapid collection and classification device, and does not consider the influence of classification on the subsequent pretreatment, enzymolysis and inhibitor of the straw.
CN 201210014796.7 discloses a method for reducing fermentation inhibitor from source, which combines steam explosion technology and air classification technology to pretreat wood fiber raw material, ensure that fiber tissue has better pretreatment effect, avoid excessive degradation of parenchyma, and improve enzymolysis rate of fiber raw material. According to the method, two sections of steam explosion are carried out on lignocellulose raw materials, classification is carried out after steam explosion pretreatment, and secondary steam explosion is carried out on fiber tissues, so that the generation of inhibitors is reduced, and the overall enzymolysis efficiency of the straw is improved. The condition used for the first step of steam explosion in the process of the method is stronger, and the energy consumption and the operation time are increased by adopting two steam explosion treatments.
CN 102586342a discloses a method for improving the tolerance of saccharomyces cerevisiae to inhibitors of cellulose hydrolysate by over-expressing ABC (ATP-binding cassette) transporter gene ADP1 to improve the resistance of saccharomyces cerevisiae to various inhibitors (formic acid, acetic acid and furfural) in cellulose hydrolysate. In order to make the thalli tolerant to the inhibitor in the cellulose hydrolysate, the method adopts a genetic engineering method to modify the saccharomyces cerevisiae, thereby improving the tolerance of the saccharomyces cerevisiae. However, in the process of utilizing cellulose hydrolysate by the strain, further detoxification treatment is required to be carried out on the hydrolysate, so that the concentration of the inhibitor in the hydrolysate is reduced.
Therefore, according to the requirements of modern industrial development and industrial equipment, based on the full cognition of straw raw materials, an engineering equipment integration is developed, the efficient uniform grading utilization of straw is realized, the straw is converted into straw sugar with low inhibitor concentration, and the method is an urgent need for large-scale sustainable development of the current biological manufacturing industry.
Disclosure of Invention
Aiming at the problems existing in the prior art, the invention provides a method for preparing low inhibitor straw sugar suitable for fermentation and catalysis by homogenizing and grading straw, which is based on the great difference of the elastic modulus of the outer skin and the medulla part of the straw, adopts the mechanochemical action to realize the homogenization and grading of the straw, obviously reduces the steam explosion pretreatment intensity in the biorefinery process, reduces substances with inhibition effect on fermentation and catalysis in the straw sugar, and improves the availability of the straw sugar; the method has the advantages of strong theoretical basis, good inhibitor reducing effect, high raw material uniformity and high treatment efficiency, and the provided process equipment of the uniform grading method is mature and has strong industrialized applicability.
In order to achieve the technical effects, the invention adopts the following technical scheme:
the invention aims to provide a method for preparing low inhibitor straw sugar suitable for fermentation and catalysis by homogenizing graded straw, which comprises the following steps:
carrying out rolling treatment on the refined straw to obtain rolled straw;
setting mechanical-chemical treatment parameters according to a mechanical grading torsion mechanical model, and carrying out mechanical grading treatment on the rolled straw by using a mechanical-chemical treatment method to obtain mechanical graded straw;
screening the mechanically graded straw to obtain graded straw;
carrying out rehydration treatment and steam explosion treatment on the graded straws to obtain steam exploded graded straws;
and carrying out high-solid enzymolysis treatment on the steam-exploded graded straw to obtain the low inhibitor straw sugar.
As a preferable technical scheme of the invention, the rolling treatment sets rolling parameters according to a rolling interval model.
Preferably, the rolling pitch model isWhere d is the roll-in pitch mm and n is the number of roll-ins.
As a preferable technical scheme of the invention, the rolling treatment device is a tooth-type rolling crushing device.
Preferably, the tooth roll crushing device comprises any one of a twin roll press, a three roll press, a tooth roll crusher or a double tooth roll crusher.
As a preferable technical scheme of the invention, the mechanical grading torsion mechanical model comprises a torsion angle model of straw skin, pi Rang adhesion tissues and pulp;
the torsion angle model of the straw skin, pi Rang adhesion tissues and pulp is as follows:
；
；
；
；
wherein the method comprises the steps of、、The elastic modulus of the straw skin, pi Rang adhesion tissue and pulp are respectively equal to R,，the radius of the skin and Pi Rang adhesion tissue and the radius of the pulp are respectively, L is the length of the straw,the torque level is mechanically graded.
Preferably, the normalized classification condition of the mechanical classification torsion mechanical model is that。
As a preferable technical scheme of the invention, the mechanical classification treatment device is a disc type grinding device.
Preferably, the disc grinding device comprises any one of a disc mill, a disc grinder, a disc refiner or a high consistency refiner.
As a preferable technical scheme of the invention, the pore diameter of the screen mesh for the grading treatment is 10-200 meshes, and the time is 1-10 min. The mesh size may be 20 mesh, 40 mesh, 60 mesh, 80 mesh, 100 mesh, 120 mesh, 160 mesh, 180 mesh, 200 mesh, or the like, and the classification time may be 1min, 1.2min, 1.5min, 1.8min, 2min, 3min, 4min, 5min, 6min, 7min, 8min, 9min, or 10min, or the like, but is not limited to the listed data, and other non-listed values within the above numerical ranges are equally applicable.
As a preferable technical scheme of the invention, the screening and grading straw is a grading straw with the elastic modulus of 0.10-0.50 MPa, such as 0.10 MPa, 0.15 MPa, 0.20 MPa, 0.25 MPa, 0.30 MPa, 0.35 MPa, 0.40 MPa, 0.45 MPa or 0.50MPa, etc., but the invention is not limited to the listed data, and other non-listed values in the numerical range are applicable.
The preferable technical scheme of the invention is characterized in that the water content of the rehydrated straw is 40-60 wt%, such as 40 wt%, 42 wt%, 45 wt%, 48 wt%, 50 wt%, 52 wt%, 55 wt%, 58 wt% or 60 wt%, etc., but the invention is not limited to the listed data, and other non-listed values in the numerical range are equally applicable.
As a preferable technical scheme of the invention, setting steam explosion parameters of the steam explosion model according to the steam explosion and inhibitor generation model;
the steam explosion and inhibitor generation model is as follows:
；
wherein W is E Is the total energy of steam explosion, ϒ is the gas insulation index, P 0 Is at standard atmospheric pressure, T i Is the initial temperature, r 0 Is the cell radius, r 1 Is the cell radius after expanding the cavity, R s Is the outer diameter of the substrate (sphere) per unit mass, R c0 For expanding the outer diameter of plant cells, sigma r Is radial stress, sigma θ Is tangential stress; r is the distance from the calculated point to the center point of the explosion source; alpha is the propagation attenuation index of blasting load, mu d For the dynamic poisson ratio of the medium, c s Is the specific heat capacity, m of the substrate s Is the mass of the substrate.
As a preferable technical scheme of the invention, the low inhibitor straw sugar is used for preparing any one of ethanol, butanol, lactic acid, 1, 3-butanediol, 2, 3-butanediol, lysine and glutathione by a fermentation method.
Preferably, the low inhibitor straw sugar is prepared by any one of ethylene glycol, sorbitol, propylene glycol, mannitol, maltitol, xylitol, furfural and 5-hydroxymethylfurfural through a catalytic fermentation method.
Compared with the prior art, the invention has at least the following beneficial effects:
(1) The method comprises the steps of establishing a steam explosion pretreatment process inhibitor generation model, carrying out theoretical analysis on a generation mechanism of inhibitors in straw sugar, carrying out classification treatment on straw raw materials according to the theoretical model, and obtaining the straw sugar capable of being fermented and catalyzed after low-strength steam explosion pretreatment and high-solid enzymolysis on the obtained classified straw;
(2) The method for preparing the straw sugar with low inhibitor, which is applicable to fermentation and catalysis, by using the homogenized and graded straw, can reduce the straw sugar inhibitor from the source and simultaneously reduce the cost and energy consumption of the biorefinery of the straw sugar;
(3) Compared with the existing inhibitor removal modes such as activated carbon adsorption, the method starts from raw materials, and builds a uniform classification method according to the mechanical property difference of the biological tissue structure of the straw, so that a classified straw component with lower elastic modulus is obtained, the inhibitor in the straw sugar is effectively reduced, and the availability of the straw sugar is improved;
(4) The method for preparing the low inhibitor straw sugar suitable for fermentation and catalysis by homogenizing and grading straw is characterized in that equipment and process required by the method are mature, mechanical grading equipment is mature, inhibitor removal cost in the straw sugar utilization process can be reduced, and fermentation utilization and catalytic conversion of the straw sugar are promoted.
Drawings
FIG. 1 shows the state of the straw after the rolling treatment in example 1 (the number of rolling times from right to left is 1 to 5);
Fig. 2 is an elastic modulus test chart of straw components with more than 10 meshes after mechanochemical treatment (the number of mechanochemical treatment times from top left to bottom right is 1 to 5) in example 1;
FIG. 3 is a graph showing the axial elastic modulus test of the graded straw after mechanochemical treatment (the number of mechanochemical treatments from top left to bottom right is 1 to 5) in example 1;
FIG. 4 is a graph showing the elastic modulus of the 10 mesh or more straw components after mechanochemical treatment in example 1;
FIG. 5 is a graph showing the axial elastic modulus test of the graded straw after mechanochemical treatment in example 1;
FIG. 6 is a photomicrograph of the graded straw composition and steam exploded graded straw (graded straw, 0.5MPa,5min steam exploded treated straw, 0.8MPa,5min steam exploded treated straw in order from left to right);
FIG. 7 is a graph showing the enzymatic hydrolysis rate of the high solids enzymatic hydrolysis process of examples 1-5 and comparative examples 1-2;
FIG. 8 is a graph comparing the steam explosion strength of different steam explosion conditions in an embodiment of the invention;
FIG. 9 is a graph showing the yield of 2, 3-butanediol from the straw carbohydrate hydrolysate provided in example 1 (wherein CSFS represents a fractionated straw hydrolysate of corn);
fig. 10 is a chart showing the catalytic conversion of sorbitol from the stalk sugar hydrolysate provided in example 1.
The present invention will be described in further detail below. The following examples are merely illustrative of the present invention and are not intended to represent or limit the scope of the invention as defined in the claims.
Detailed Description
The technical scheme of the invention is further described below by the specific embodiments with reference to the accompanying drawings.
The invention provides a method for preparing low inhibitor straw sugar suitable for fermentation and catalysis by homogenizing graded straw, which comprises the following steps:
carrying out rolling treatment on the refined straw to obtain rolled straw;
setting mechanical-chemical treatment parameters according to a mechanical grading torsion mechanical model, and carrying out mechanical grading treatment on the rolled straw by using a mechanical-chemical treatment method to obtain mechanical graded straw;
screening the mechanically graded straw to obtain graded straw;
carrying out rehydration treatment and steam explosion treatment on the graded straws to obtain steam exploded graded straws;
and carrying out high-solid enzymolysis treatment on the steam-exploded graded straw to obtain the low inhibitor straw sugar.
Compared with the existing mechanical separation method and inhibitor reduction method, the homogenization grading method provided by the invention is based on physical characteristic cognition of straw raw materials, and an energy model and an inhibitor generation model of the straw pretreatment process are constructed. The invention provides a homogenization and grading method, which takes a homogenization and grading torsion mechanical model as theoretical guidance, and adopts mechanochemical action aiming at the great difference of elastic modulus of the outer skin and the medulla part of the straw to change the organization, the structure and the performance of the straw so as to realize the homogenization and grading of the straw. In the biorefinery process, the method is coupled with steam explosion pretreatment and high-solid enzymolysis, so that the steam explosion pretreatment intensity can be remarkably reduced, the concentration of inhibitors in the straw enzymolysis liquid is reduced, the straw biorefinery energy consumption is reduced, the availability of the straw sugar liquid is improved, and a rich renewable sugar source is provided for the modern biological manufacturing and chemical catalysis industries.
In one embodiment of the present invention, the straw comprises any one or a combination of at least two of corn straw, rice straw, wheat straw, or sorghum straw, typical but non-limiting examples of which are: a combination of corn stalk and rice stalk, a combination of rice stalk and wheat stalk, a combination of wheat stalk and sorghum stalk, a combination of corn stalk, rice stalk and wheat stalk, or a combination of corn stalk, rice stalk, wheat stalk and sorghum stalk, etc.
In one specific embodiment of the invention, the refining treatment comprises leaf removal, dust removal, soil removal and the like, so that impurity parts in the straws are removed, and the damage of the impurities to mechanical equipment is reduced.
In one embodiment of the present invention, the straw is cut into small pieces of about 10 to 30cm after the thinning treatment, such as 10cm, 10.5cm, 11cm, 13cm, 15cm, 17cm, 19cm, 20cm, 22cm, 24cm, 26cm, 28cm, 29cm or 30cm, etc., but not limited to the listed data, and other non-listed values within the numerical range are equally applicable.
In a specific embodiment of the present invention, the rolling treatment device may be a roll press, a three-roll press, a toothed roll crusher, a double-toothed roll crusher, or other common toothed roll crushing devices.
In one embodiment of the present invention, the number of times of the rolling treatment may be 1 to 10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 times.
In one embodiment of the present invention, the apparatus of the mechanochemical treatment method may be a plurality of common disc grinding apparatuses such as a disc mill, a disc grinder, a disc refiner or a high consistency refiner.
In one specific embodiment of the invention, the relation between the times n of the mechanochemical treatment method and the distance D (mm) between the grinding discs is as follows:wherein n may be 1 to 10 times, such as 1 time, 2 times, 3 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times or 10 times.
In one embodiment of the invention, the screening treatment equipment can be a high-frequency vibrating screen, a linear vibrating screen, a rotary vibrating screen or a circular vibrating screen and other common screening equipment.
According to the invention, the main effect of the screening treatment is to separate the graded straws with different particle sizes obtained after mechanical grading, the graded straw components have small elastic modulus and low mechanical strength, the graded straw components are crushed into fine particles in the mechanical grading process, the elastic modulus of the straw skin components is large, the mechanical strength is high, and the straw skin components are kept in a long strip shape in the mechanical grading process, so that the size of the screen mesh can be selected to be 20-200 meshes.
In the invention, the mechanically graded straw components are subjected to rehydration treatment, so that water fully enters the interior of cells of the graded straw components, then steam explosion pretreatment is performed, the cell walls of the graded straw components are damaged in the high-temperature steam cooking and instantaneous explosion processes, the compact structures of cellulose, hemicellulose and lignin are damaged, and more binding sites are provided for the combination of cellulase in the enzymolysis process. The mechanical graded straw component has low elastic modulus and weak mechanical strength, the pretreatment strength required by the compact structure of the graded straw component to be destroyed in the steam explosion pretreatment process is lower, and the inhibitory compounds such as organic acid, furan compounds, phenolic acid compounds and the like generated in the steam explosion pretreatment process are lower. The grading straw component obtained by the method can obviously reduce the strength of steam explosion pretreatment and obviously reduce the concentration of inhibitors in straw enzymolysis liquid.
In one embodiment of the present invention, the steam explosion parameters of the steam explosion model set by the steam explosion and inhibitor generation model can be shown in table 1;
TABLE 1
The calculation method of the steam explosion strength is as follows;
wherein T is the treatment time and T is the treatment temperature;
According to the energy model and the inhibitor generation model, the steam explosion strength is reduced from 3841 to 1133.6, the energy consumed by the inhibitor in the steam explosion process is reduced by 15.16%, and when the energy is reduced to 238.4, the concentration of the inhibitor is reduced by 28.88%; meanwhile, when the steam explosion temperature is reduced from 453K to 378K, the formic acid yield is reduced by 46.6%, the acetic acid yield is reduced by 59.2%, the 5-hydroxymethylfurfural yield is reduced by 20.7%, and the furfural yield is reduced by 176.9%; according to the generation model of the inhibitor in the steam explosion process, the raw material elasticity model is increased by 2 times, the formic acid generation amount is increased by 113.6%, the acetic acid generation amount is increased by 111.5%, and the 5-hydroxymethylfurfural generation amount is increased by 65.2%.
In one embodiment of the present invention, in order to reduce the concentration of the inhibitor generated during the steam explosion, the pressure of the steam explosion treatment is 0.4 to 0.8Mpa, such as 0.4Mpa, 0.45Mpa, 0.5Mpa, 0.55Mpa, 0.6Mpa, 0.65Mpa, 0.7Mpa, 0.75Mpa or 0.8Mpa, etc.; the pressure maintaining time of the steam explosion treatment is 2-8 min, such as 2min, 3min, 4min, 5min, 6min, 7min or 10min, but is not limited to the listed data, and other non-listed values in the above numerical ranges are equally applicable.
In one embodiment of the invention, the method for preparing the low inhibitor straw sugar by enzymolysis treatment can be high-solid enzymolysis.
In a specific embodiment of the invention, the high-solid enzymolysis condition can be that the solid content is 15-25% (w/w), the cellulase dosage is 5-20 FPU/g DM (absolute dry material), the pH is 4.0-5.0, and the enzymolysis is carried out for 72 hours at 50 ℃. Wherein the solid content can be 15wt%, 16wt%, 17wt%, 18wt%, 19wt%, 20wt%, 21wt%, 22wt%, 23wt%, 24wt%, 25wt%, etc.; the cellulase may be used in an amount of 5FPU/g DM, 6FPU/g DM, 7FPU/g DM, 10FPU/g DM, 12FPU/g DM, 14FPU/g DM, 16FPU/g DM, 18FPU/g DM or 20FPU/g DM, etc., but is not limited to the enumerated values, as other non-enumerated values within each of the aforementioned numerical ranges are equally applicable. An environment of pH 4.0 to 5.0 may be provided by a citric acid-sodium citrate buffer solution (0.05 mol/L, ph=4.8).
In one specific embodiment of the invention, the high-solid enzymolysis is performed in a constant-temperature enzymolysis device, and the constant-temperature enzymolysis device can be a common enzymolysis reaction device such as a constant-temperature incubator, a constant-temperature shaking table or a constant-temperature stirring tank.
For a better illustration of the present invention, which is convenient for understanding the technical solution of the present invention, exemplary but non-limiting examples of the present invention are as follows:
example 1
The embodiment provides a method for preparing low inhibitor straw sugar suitable for fermentation and catalysis by homogenizing graded straw, which comprises the following steps:
(1) Cutting corn stalk into 10cm sections;
(2) Setting parameters according to the relation between the rolling interval and the times, and rolling the cut corn straw for 3 times by using a tooth type three-roller rolling machine to obtain rolled corn straw;
(3) According to the relation between disc grinding interval and times, disc grinding is carried out on the rolled corn straw for 2 times by using a disc mill (rated power is 3500W, rotating speed is 1500 rpm) to obtain the corn straw subjected to mechanochemical treatment, a high-frequency vibrating screen is used for mechanically classifying the corn straw, the mechanical classifying screen is used for screening for 2 minutes, classified straw components with particle sizes below 40 meshes are obtained, and after elastic modulus experiment test, the elastic modulus of the classified straw is about 0.16Mpa;
(4) Adding deionized water into the graded straw components for rehydration, adjusting the water content of the graded straw components to 50wt%, and then placing the rehydrated graded straw components into a steam explosion reaction device, and maintaining for 5min under the steam explosion pressure of 0.5 MPa; obtaining a steam explosion grading straw component;
(5) Collecting steam-exploded graded straw components, adding a citric acid-sodium citrate buffer solution (0.05M, pH=4.8), regulating the solid content to 25%, adding cellulase (10 FPU/g DM) into the mixture, fully mixing the materials with the cellulase, and placing the mixture into a constant-temperature shaking table at 50 ℃ for enzymolysis reaction to obtain the straw enzymolysis sugar solution.
Example 2
The embodiment provides a method for preparing low inhibitor straw sugar suitable for fermentation and catalysis by homogenizing graded straw, which comprises the following steps:
(1) Cutting corn stalk into 20cm sections;
(2) Setting parameters according to the relation between the rolling interval and the times, and rolling the cut corn straw for 4 times by using a tooth type three-roller rolling machine to obtain rolled corn straw;
(3) According to the relation between disc grinding interval and times, disc grinding is carried out on the rolled corn straws for 2 times by using a disc mill (rated power is 3500W, rotating speed is 1500 rpm) to obtain corn straws subjected to mechanochemical treatment, a high-frequency vibrating screen is used for mechanically grading the corn straws, and after screening is carried out on a 40-mesh screen for 2 minutes, graded straws with grain diameters below 40 meshes are obtained, and the elastic modulus is about 0.17Mpa;
(4) Adding deionized water into the graded straw components for rehydration, adjusting the water content of the graded straw components to 50wt%, and then placing the rehydrated graded straw components into a steam explosion reaction device, and maintaining for 5min under the steam explosion pressure of 0.8 MPa; obtaining a steam explosion grading straw component;
(5) Collecting steam-exploded graded straw components, adding a citric acid-sodium citrate buffer solution (0.05M, pH=4.8), regulating the solid content to 25%, adding cellulase (10 FPU/g DM) into the mixture, fully mixing the materials with the cellulase, and placing the mixture into a constant-temperature shaking table at 50 ℃ for enzymolysis reaction to obtain the straw enzymolysis sugar solution.
Example 3
The embodiment provides a method for preparing low inhibitor straw sugar suitable for fermentation and catalysis by homogenizing graded straw, which comprises the following steps:
(1) Cutting corn stalk into 30cm sections;
(2) Setting parameters according to the relation between the rolling interval and the times, and rolling the cut corn straw for 4 times by using a tooth type three-roller rolling machine to obtain rolled corn straw;
(3) According to the relation between disc grinding interval and times, disc grinding is carried out on the rolled corn straws for 2 times by using a disc mill (rated power is 3500W, rotating speed is 1500 rpm) to obtain corn straws subjected to mechanochemical treatment, a high-frequency vibrating screen is used for the mechanically graded corn straws, and after screening is carried out on a 20-mesh screen for 2 minutes, graded straws with grain diameters below 40 meshes are obtained, and the elastic modulus is about 0.18Mpa;
(4) Adding deionized water into the graded straw components for rehydration, adjusting the water content of the graded straw components to 50wt%, and then placing the rehydrated graded straw components into a steam explosion reaction device, and maintaining for 5min under the steam explosion pressure of 0.5 MPa; obtaining a steam explosion grading straw component;
(5) Collecting steam-exploded graded straw components, adding a citric acid-sodium citrate buffer solution (0.05M, pH=4.8), regulating the solid content to 25%, adding cellulase (10 FPU/g DM) into the mixture, fully mixing the materials with the cellulase, and placing the mixture into a constant-temperature shaking table at 50 ℃ for enzymolysis reaction to obtain the straw enzymolysis sugar solution.
Example 4
The embodiment provides a method for preparing low inhibitor straw sugar suitable for fermentation and catalysis by homogenizing graded straw, which comprises the following steps:
(1) Dedusting and impurity-removing corn straw, and cutting the corn straw into small sections of 30 cm;
(2) Setting parameters according to the relation between the rolling interval and the times, and rolling the cut corn straw for 4 times by using a tooth type three-roller rolling machine to obtain rolled corn straw;
(3) According to the relation between disc grinding interval and times, disc grinding is carried out on the rolled corn straw for 2 times by using a disc mill (rated power is 3500W, rotating speed is 1500 rpm) to obtain the corn straw subjected to mechanochemical treatment, a high-frequency vibrating screen is used for the mechanically graded corn straw, and after screening is carried out on a 20-mesh screen for 2 minutes, a graded straw component with the grain diameter of below 40 meshes is obtained, and the elastic modulus is about 0.17Mpa;
(4) Adding deionized water into the graded straw components for rehydration, adjusting the water content of the graded straw components to 50wt%, and then placing the rehydrated graded straw components into a steam explosion reaction device, and maintaining for 5min under the steam explosion pressure of 0.8 MPa; obtaining a steam explosion grading straw component;
(5) Collecting steam-exploded graded straw components, adding a citric acid-sodium citrate buffer solution (0.05M, pH=4.8), regulating the solid content to 25%, adding cellulase (10 FPU/g DM) into the mixture, fully mixing the materials with the cellulase, and placing the mixture into a constant-temperature shaking table at 50 ℃ for enzymolysis reaction to obtain the straw enzymolysis sugar solution.
Example 5
The embodiment provides a method for preparing low inhibitor straw sugar suitable for fermentation and catalysis by homogenizing graded straw, which comprises the following steps:
(1) After the corn stalk is dedusted and decontaminated, cutting the corn stalk into small sections of 30 cm;
(2) Setting parameters according to the relation between the rolling interval and the times, and rolling the cut corn straw for 2 times by using a tooth type three-roller rolling machine to obtain rolled corn straw;
(3) According to the relation between disc grinding interval and times, disc grinding is carried out on the rolled corn straws for 2 times by using a disc mill (rated power is 3500W, rotating speed is 1500 rpm) to obtain corn straws subjected to mechanochemical treatment, a high-frequency vibrating screen is used for mechanically grading the corn straws, and after screening is carried out on a 40-mesh screen for 2 minutes, graded straws with grain diameters below 40 meshes are obtained, and the elastic modulus is about 0.18Mpa;
(4) Adding deionized water into the graded straw components for rehydration, adjusting the water content of the graded straw components to 50wt%, and then placing the rehydrated graded straw components into a steam explosion reaction device, and maintaining for 5min under the steam explosion pressure of 0.5 MPa; obtaining a steam explosion grading straw component;
(5) Collecting steam-exploded graded straw components, adding a citric acid-sodium citrate buffer solution (0.05M, pH=4.8), regulating the solid content to 20%, adding cellulase (20 FPU/g DM) into the mixture, fully mixing the materials with the cellulase, and placing the mixture into a constant-temperature shaking table at 50 ℃ for enzymolysis reaction to obtain the straw enzymolysis sugar solution.
Comparative example 1
This comparative example provides a method of preparing a straw saccharide, the method comprising the steps of:
(1) Dedusting and impurity-removing corn straw, and cutting into small sections of 10 cm;
(2) Adding deionized water into the graded straw components of the corns for rehydration, adjusting the water content of the straw to 50wt%, and then placing the graded straw components after rehydration into a steam explosion reaction device, and maintaining for 5min under the steam explosion pressure of 1.0 MPa; obtaining steam exploded straw;
(3) Collecting steam-exploded graded straw components, adding a citric acid-sodium citrate buffer solution (0.05M, pH=4.8), regulating the solid content to 25%, adding cellulase (10 FPU/g DM) into the mixture, fully mixing the materials with the cellulase, and placing the mixture into a constant-temperature shaking table at 50 ℃ for enzymolysis reaction to obtain the straw enzymolysis sugar solution.
Comparative example 2
This comparative example provides a method of preparing a straw saccharide, the method comprising the steps of:
(1) Dedusting and impurity-removing corn straw, and cutting the corn straw into small sections of 30 cm;
(2) Rolling the cut corn straw for 4 times by using a tooth type three-roller press to obtain rolled corn straw;
(3) The corn stalks after the double rolling are subjected to disc grinding for 2 times by using a disc mill (rated power is 3500W, rotating speed is 1500 rpm) to obtain corn stalks after the mechanochemical treatment, the mechanically graded corn stalks are subjected to high-frequency vibration sieve, and after sieving for 2min on a 40-mesh sieve, the elastic modulus of the stalk components above the 40-mesh sieve is about 13.07Mpa;
(4) Adding deionized water into straw components with more than 10 meshes for rehydration, adjusting the water content of the graded straw components to 50wt%, then placing the rehydrated graded straw components into a steam explosion reaction device, and maintaining for 5min under the steam explosion pressure of 1.0 MPa; obtaining straw components with the size of more than 10 meshes of steam explosion;
(5) Collecting straw components with the size of more than 10 meshes after steam explosion, adding a citric acid-sodium citrate buffer solution (0.05M, pH=4.8), regulating the solid content to 25%, adding cellulase (10 FPU/g DM) into the mixture, fully mixing the materials with the cellulase, and placing the mixture into a constant-temperature shaking table at 50 ℃ for enzymolysis reaction to obtain the straw enzymolysis sugar solution.
The inhibitor concentrations in the stalk enzymatic sugar solutions obtained in examples 1 to 5 and comparative examples 1 to 2 were tested, and the results are shown in Table 2. The specific test method comprises the following steps: centrifuging the enzymolysis solution at 10000 rpm for 10 min. And taking the supernatant after centrifugation, passing through a 0.22 mu m filter membrane, and measuring the concentration of glucose, xylose, formic acid, acetic acid, furfural and 5-HMF by adopting a high performance liquid chromatography. Chromatographic conditions: HPX-87H ion exchange column, mobile phase is 5 mM sulfuric acid, flow rate is 0.6 mL/min; the detector temperature and column temperature were 35 ℃ and 40 ℃, respectively.
TABLE 2
As can be seen from Table 2, the relative concentrations of the three representative inhibitors in examples 1-5 averaged 4.05 g/L/100g/L total sugar, which was reduced by a factor of about 2.5 compared to the comparative example.
The elastic modulus of the straw component with more than 10 meshes after the mechanochemical treatment in the example 1 is tested, the result is shown in figure 2, and the relation between the number of times of the mechanochemical treatment and the elastic modulus is shown in figure 4; the axial elastic modulus of the graded straw after mechanochemical treatment is tested, the result is shown in figure 3, and the relation between the number of mechanochemical treatments and the axial elastic modulus is shown in figure 5. The specific detection method comprises the following steps: the elastic modulus of the straw and fractionated straw was measured by a texture analyzer (Brookfield CT3 4500). The brittleness of different tissues of the straw is measured by adopting a TA11/1000 probe, the texture performance of the raw materials before and after steam explosion is measured by adopting a TTAA36 probe, before testing, the pre-crushed classified steam explosion materials are weighed in a small 50ml beaker, the loading height is kept to be 10 mm, the probe moves downwards at a constant speed of 2 mm/s, the contact is set to be 0.2N, and the testing distance is 2 mm.
Wherein E represents axial elastic modulus (MPa) of the straw; f represents the axial acting force (N) applied to the straw; s represents the stress area (m 2 ) The method comprises the steps of carrying out a first treatment on the surface of the L represents the thickness (m) of the straw; k represents the bulk modulus (MPa) of the straw staple fiber; p represents acting force (MPa) applied to the straw short fibers; v represents the volume (m) 3 ）。
The enzymatic hydrolysis rates of the straw enzymatic hydrolysis sugar solutions obtained in examples 1 to 5 and comparative examples 1 to 2 were measured, and the results are shown in FIG. 7. The specific calculation method comprises the following steps: measuring the content of glucose in the enzymolysis liquid by high performance liquid chromatography, and then measuring the content of cellulose in the raw materials by using a standard NREL (National Renewable Energy Laboratory) method; the enzymolysis rate calculation formula is:
in the middle ofRepresents the glucose concentration in the enzymatic hydrolysate;representing the mass of cellulose in the feedstock;representing the volume of the enzymatic hydrolysate, 152/180 is the theoretical coefficient of conversion of glucose to dextran.
The results of comparing the steam explosion strengths of example 1 and comparative example 2 are shown in fig. 8.
The straw enzymolysis sugar solution obtained in the example 1 is used for fermenting 2, 3-butanediol, and the conditions are as follows:
(1) The graded straw enzymolysis liquid and the glucose solution are prepared according to 2 ml/L of fermentation medium (glucose 30 g/L (if no special description exists), yeast extract 13.33 g/L, tryptone 13.33 g/L, sodium acetate 0.67 g/L, trace element mother liquor (magnesium sulfate heptahydrate 0.68 g, manganese sulfate monohydrate 0.372 g, sodium chloride 0.333 g, ferrous sulfate heptahydrate 0.608 g, dissolved in 50 mL distilled water) and pH is adjusted to 6.0 after being fully mixed;
(2) And (3) selecting single bacterial colonies in the activation culture medium, inoculating the single bacterial colonies to a seed culture medium, activating the single bacterial colonies at 50 ℃ for 24 h, and then respectively inoculating seed solutions to a fermentation culture medium taking glucose and corn straw fermentable sugar as carbon sources. The inoculation amount is 10%, the culture condition is 50 ℃, and the rotating speed is 150 rpm;
(3) After 24. 24 h activation of the seed solution at 50℃the seed solution was inoculated into a fermentation medium having a sugar concentration of 60 g/L. The inoculum size was 10%, and 36 h was cultured at 50℃and 150 rpm. Cell density, substrate and product concentration were measured at interval 3 h.
The results of 2, 3-butanediol fermentation in pure glucose medium are shown in FIG. 9.
The straw enzymolysis sugar solution obtained in the example 1 is used for electrochemical catalytic conversion of sorbitol, and the conditions are as follows:
(1) Respectively taking the graded straw enzymolysis liquid and a sodium sulfate solution of glucose as a catholyte, taking a 10% sodium sulfate solution as an anolyte, taking a ruthenium iridium titanium electrode as an anode, and taking foam nickel containing Raney nickel as a cathode to construct an H-type electrolytic cell;
(2) Placing the electrolytic cell in 50deg.C water bath, connecting with electrochemical workstation, and introducing 5mA/cm 2 And (3) carrying out electrocatalytic operation on the constant current, sampling and detecting the mixed solution in the constant current at intervals of 30min, and observing the electrocatalytic conditions of the enzymolysis solution system and the glucose solution system.
The results of electrochemical catalytic conversion of sorbitol in pure glucose medium are shown in FIG. 10.
From fig. 1, it can be seen that after the straw is subjected to the sectional treatment, the three-roller tooth type roller press is used for rolling for 1-5 times, the rolling times can be seen to be increased, the crushing degree of the straw is increased, and the formed preliminary graded straw is reduced, so that the overall hardness of the straw is reduced, and the follow-up mechanical grading is more facilitated.
As can be seen from fig. 2 and 4, the elastic modulus of the straw component with more than 10 meshes is reduced from 13.90MPa to 9.49MPa along with the increase of the mechanical classification times, and the mechanical classification causes a certain damage to the densification mechanism of the straw component, so that the reduction is small.
As can be seen from fig. 3 and 5, the elastic modulus of the graded straw increased from 0.12MPa to 0.41MPa with increasing number of mechanical grading. It can be seen from fig. 4 and 5 that after mechanical classification, the elastic modulus of the straw is about 10MPa, and the elastic modulus of the classified straw is about 0.20-0.30 MPa, which is different by more than 50 times, so that the classified straw with huge elastic modulus in the corn straw can be effectively classified by mechanical classification, thereby being beneficial to subsequent biorefinery and biological utilization.
From fig. 6, it can be seen that the classified straw is in a fine spherical structure, the surface is loose and porous, and basically presents a broken state after being subjected to low-strength steam explosion pretreatment of 0.5mpa and 5min, which is more beneficial to the subsequent enzymolysis reaction, and also indicates that the steam explosion pretreatment strength required by the classified straw after mechanical classification is low.
From fig. 7, it can be seen that in examples 1 to 5, the mechanically classified straw was pretreated under the steam explosion conditions of 0.5mpa,5min or 0.8mpa,5min, whereas the non-classified straw and the classified straw component of comparative example were steam-exploded under the steam explosion conditions of 1.0mpa,5min, and then subjected to the enzymatic hydrolysis reaction under the same enzymatic hydrolysis conditions, it can be seen that the enzymatic hydrolysis rate of the classified straw component was about 58%, the enzymatic hydrolysis rate of comparative example 1 was about 42%, the enzymatic hydrolysis rate of the skin component of comparative example 2 was about 26%, and the enzymatic hydrolysis performance of the classified straw after the low-strength steam explosion pretreatment was about 40% higher than that of the non-classified straw by more than 2 times. Further illustrating the necessity of mechanical classification of the straw.
As can be seen from FIG. 8, the 0.5MPa,5min steam explosion treatment strength in example 1 was 255.10, the 1.0MPa,5min steam explosion treatment strength in comparative example 2 was 1125.96, and the steam explosion strength in example 1 was reduced by 4.41 times than that in comparative example 2.
As can be seen from FIG. 9, the 2, 3-butanediol obtained by fermenting the 2, 3-butanediol by using the graded straw enzymolysis liquid reaches 13.80g/L, and the yield is increased by 13.02 percent compared with the yield of 12.21g/L of the pure glucose culture medium.
From FIG. 10, it can be seen that the concentration of sorbitol reaches 6.05g/L and the concentration of pure glucose solution reaches 7.66g/L after 3h catalytic conversion, which reaches about 78.89% of the catalysis of the pure glucose solution.
The applicant states that the detailed structural features of the present invention are described by the above embodiments, but the present invention is not limited to the above detailed structural features, i.e. it does not mean that the present invention must be implemented depending on the above detailed structural features. It should be apparent to those skilled in the art that any modifications of the present invention, equivalent substitutions of selected components of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope of the present invention and the scope of the disclosure.
The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited to the specific details of the above embodiments, and various simple modifications can be made to the technical solution of the present invention within the scope of the technical concept of the present invention, and all the simple modifications belong to the protection scope of the present invention.
In addition, the specific features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, various possible combinations are not described further.
Moreover, any combination of the various embodiments of the invention can be made without departing from the spirit of the invention, which should also be considered as disclosed herein.
Claims (7)
1. A method for preparing low inhibitor straw sugars suitable for fermentation and catalysis by homogenizing graded straw, the method comprising the steps of:
(1) Cutting corn stalk into 10cm sections;
(2) Setting parameters according to the relation between the rolling interval and the times, and rolling the cut corn straw for 3 times by using a tooth type three-roller rolling machine to obtain rolled corn straw;
the relation between the rolling interval and the times is d=1.6-0.1 n, wherein d is the rolling interval mm, and n is the rolling times;
(3) According to the relation between disc grinding interval and times, using a disc mill for the corn straw after the pair rolling, wherein the rated power is 3500W, the rotating speed is 1500 r/min, obtaining the corn straw after the mechanochemical treatment after disc grinding for 2 times, using a high-frequency vibrating screen for the corn straw after mechanical classification, screening for 2min on a 40-mesh screen, obtaining a classified straw component with the grain diameter of less than 40 meshes, and testing the elasticity modulus of the classified straw by an elasticity modulus experiment to be 0.16Mpa;
The relation formula of the number of times n of disc grinding and the distance D between the grinding discs is as follows: d=1.1-0.1 n, units mm of D;
(4) Adding deionized water into the graded straw components for rehydration, adjusting the water content of the graded straw components to 50wt%, and then placing the rehydrated graded straw components into a steam explosion reaction device, and maintaining for 5min under the steam explosion pressure of 0.5 MPa; obtaining a steam explosion grading straw component;
(5) Collecting steam-exploded graded straw components, adding 0.05M citric acid-sodium citrate buffer solution, adjusting pH to 4.8, adjusting solid content to 25%, adding 10FPU/g DM cellulase, fully mixing the materials with the cellulase, and placing the mixture into a constant temperature shaking table at 50 ℃ for enzymolysis reaction to obtain the straw enzymolysis sugar solution.
2. A method for preparing low inhibitor straw sugars suitable for fermentation and catalysis by homogenizing graded straw, the method comprising the steps of:
(1) Cutting corn stalk into 20cm sections;
(2) Setting parameters according to the relation between the rolling interval and the times, and rolling the cut corn straw for 4 times by using a tooth type three-roller rolling machine to obtain rolled corn straw;
the relation between the rolling interval and the times is d=1.6-0.1 n, wherein d is the rolling interval mm, and n is the rolling times;
(3) According to the relation between disc grinding interval and times, using a disc mill for the corn straw after the pair rolling, wherein the rated power is 3500W, the rotating speed is 1500 r/min, obtaining the corn straw after the mechanochemical treatment after disc grinding for 2 times, using a high-frequency vibrating screen for the corn straw after mechanical classification, screening the corn straw on a 40-mesh screen for 2min, obtaining the classified straw with the grain diameter below 40 meshes, and the elastic modulus is 0.17Mpa;
the relation formula of the number of times n of disc grinding and the distance D between the grinding discs is as follows: d=1.1-0.1 n, units mm of D;
(4) Adding deionized water into the graded straw components for rehydration, adjusting the water content of the graded straw components to 50wt%, and then placing the rehydrated graded straw components into a steam explosion reaction device, and maintaining for 5min under the steam explosion pressure of 0.8 MPa; obtaining a steam explosion grading straw component;
(5) Collecting steam-exploded graded straw components, adding 0.05M citric acid-sodium citrate buffer solution, adjusting pH to 4.8, adjusting solid content to 25%, adding 10FPU/g DM cellulase, fully mixing the materials with the cellulase, and placing the mixture into a constant temperature shaking table at 50 ℃ for enzymolysis reaction to obtain the straw enzymolysis sugar solution.
3. A method for preparing low inhibitor straw sugars suitable for fermentation and catalysis by homogenizing graded straw, the method comprising the steps of:
(1) Cutting corn stalk into 30cm sections;
(2) Setting parameters according to the relation between the rolling interval and the times, and rolling the cut corn straw for 4 times by using a tooth type three-roller rolling machine to obtain rolled corn straw;
the relation between the rolling interval and the times is d=1.6-0.1 n, wherein d is the rolling interval mm, and n is the rolling times;
(3) According to the relation between disc grinding interval and times, using a disc mill for the corn straw after the pair rolling, wherein the rated power is 3500W, the rotating speed is 1500 r/min, obtaining the corn straw after the mechanochemical treatment after disc grinding for 2 times, using a high-frequency vibrating screen for the corn straw after mechanical classification, screening the corn straw on a 20-mesh screen for 2min, obtaining the classified straw with the grain diameter of below 40 meshes, and the elastic modulus is 0.18Mpa;
the relation formula of the number of times n of disc grinding and the distance D between the grinding discs is as follows: d=1.1-0.1 n, units mm of D;
(4) Adding deionized water into the graded straw components for rehydration, adjusting the water content of the graded straw components to 50wt%, and then placing the rehydrated graded straw components into a steam explosion reaction device, and maintaining for 5min under the steam explosion pressure of 0.5 MPa; obtaining a steam explosion grading straw component;
(5) Collecting steam-exploded graded straw components, adding 0.05M citric acid-sodium citrate buffer solution, adjusting pH to 4.8, adjusting solid content to 25%, adding 10FPU/g DM cellulase, fully mixing the materials with the cellulase, and placing the mixture into a constant temperature shaking table at 50 ℃ for enzymolysis reaction to obtain the straw enzymolysis sugar solution.
4. A method for preparing low inhibitor straw sugars suitable for fermentation and catalysis by homogenizing graded straw, the method comprising the steps of:
(1) Dedusting and impurity-removing corn straw, and cutting the corn straw into small sections of 30 cm;
(2) Setting parameters according to the relation between the rolling interval and the times, and rolling the cut corn straw for 4 times by using a tooth type three-roller rolling machine to obtain rolled corn straw;
the relation between the rolling interval and the times is d=1.6-0.1 n, wherein d is the rolling interval mm, and n is the rolling times;
(3) According to the relation between disc grinding interval and times, using a disc mill for the corn straw after the pair rolling, wherein the rated power is 3500W, the rotating speed is 1500 r/min, obtaining the corn straw after the mechanochemical treatment after disc grinding for 2 times, using a high-frequency vibrating screen for the corn straw after mechanical classification, screening the corn straw on a 20-mesh screen for 2min, obtaining a classified straw component with the grain diameter of below 40 meshes, and the elastic modulus is 0.17Mpa;
the relation formula of the number of times n of disc grinding and the distance D between the grinding discs is as follows: d=1.1-0.1 n, units mm of D;
(4) Adding deionized water into the graded straw components for rehydration, adjusting the water content of the graded straw components to 50wt%, and then placing the rehydrated graded straw components into a steam explosion reaction device, and maintaining for 5min under the steam explosion pressure of 0.8 MPa; obtaining a steam explosion grading straw component;
(5) Collecting steam-exploded graded straw components, adding 0.05M citric acid-sodium citrate buffer solution, adjusting pH to 4.8, adjusting solid content to 25%, adding 10FPU/g DM cellulase, fully mixing the materials with the cellulase, and placing the mixture into a constant temperature shaking table at 50 ℃ for enzymolysis reaction to obtain the straw enzymolysis sugar solution.
5. A method for preparing low inhibitor straw sugars suitable for fermentation and catalysis by homogenizing graded straw, the method comprising the steps of:
(1) After the corn stalk is dedusted and decontaminated, cutting the corn stalk into small sections of 30 cm;
(2) Setting parameters according to the relation between the rolling interval and the times, and rolling the cut corn straw for 2 times by using a tooth type three-roller rolling machine to obtain rolled corn straw;
the relation between the rolling interval and the times is d=1.6-0.1 n, wherein d is the rolling interval mm, and n is the rolling times;
(3) According to the relation between disc grinding interval and times, using a disc mill for the corn straw after the pair rolling, wherein the rated power is 3500W, the rotating speed is 1500 r/min, obtaining the corn straw after the mechanochemical treatment after disc grinding for 2 times, using a high-frequency vibrating screen for the corn straw after mechanical classification, screening the corn straw on a 40-mesh screen for 2min, obtaining the classified straw with the grain diameter below 40 meshes, and the elastic modulus is 0.18Mpa;
The relation formula of the number of times n of disc grinding and the distance D between the grinding discs is as follows: d=1.1-0.1 n, units mm of D;
(4) Adding deionized water into the graded straw components for rehydration, adjusting the water content of the graded straw components to 50wt%, and then placing the rehydrated graded straw components into a steam explosion reaction device, and maintaining for 5min under the steam explosion pressure of 0.5 MPa; obtaining a steam explosion grading straw component;
(5) Collecting steam-exploded graded straw components, adding 0.05M citric acid-sodium citrate buffer solution with pH value of 4.8, regulating the solid content to 20%, adding 20FPU/g DM cellulase, fully mixing the materials with the cellulase, and placing the mixture into a constant temperature shaking table at 50 ℃ for enzymolysis reaction to obtain the straw enzymolysis sugar solution.
6. The method according to any one of claims 1 to 5, wherein the straw enzymatic hydrolysate is prepared by a fermentation method to any one of ethanol, butanol, lactic acid, 1, 3-butanediol, 2, 3-butanediol, lysine and glutathione.
7. The method according to any one of claims 1 to 5, wherein the straw enzymatic hydrolysate is prepared by a catalytic fermentation process to any one of ethylene glycol, sorbitol, propylene glycol, mannitol, maltitol, xylitol, furfural and 5-hydroxymethylfurfural.
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