CN113862318B - Lignocellulose refining system and method - Google Patents

Lignocellulose refining system and method Download PDF

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CN113862318B
CN113862318B CN202111144783.7A CN202111144783A CN113862318B CN 113862318 B CN113862318 B CN 113862318B CN 202111144783 A CN202111144783 A CN 202111144783A CN 113862318 B CN113862318 B CN 113862318B
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lignin
fermentation
lignocellulose
raffinate
tank
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CN113862318A (en
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张栩
张立鹤
王迁
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Beijing University of Chemical Technology
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Beijing University of Chemical Technology
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Abstract

The invention relates to a fermentable lignocellulose pretreatment agent, which comprises a carbon source, inorganic acid and betaine. The invention also relates to a green efficient refining system and method for pretreating lignocellulose based on the fermentable solvent. The system and the method comprise a full resource utilization scheme of lignocellulose, realize green and efficient refining of all components of lignin, hemicellulose and cellulose, provide a new idea of green and efficient utilization of lignocellulose resources, overcome the defect of difficult recovery and treatment of extraction solvents, effectively reduce the cost of the process and expand the processing industry chain of the lignocellulose resources. Is helpful to promote the comprehensive utilization level of renewable lignocellulose resources in China. Has great value for large-scale industrial production and practical application.

Description

Lignocellulose refining system and method
Technical Field
The invention belongs to the fields of biochemical engineering and biomass energy, relates to a lignocellulose refining system and method, and in particular relates to a lignocellulose green efficient refining system and method based on a fermentable pretreatment agent.
Background
Today facing resource crisis and environmental protection crisis in the world, how to solve the dependence on fossil resources and find renewable biomass resource utilization schemes is a great problem concerning energy strategy and economic development in China. The existing energy utilization structure and the traditional chemical industry technology need to be upgraded and improved, and the biorefinery process mainly comprising lignocellulose raw materials is one of the important ways for realizing traditional industrial substitution, energy conservation, consumption reduction and emission reduction.
Lignocellulose, the most abundant renewable biomass resource worldwide, has great potential in biofuel production and in the production of biologicals. However, lignocellulose, which is mainly a plant resource, forms complex chemical components and structures in a long process formed in the growth process, and lignin, hemicellulose and cellulose form a refractory resistance barrier, so that microorganisms, enzymes and the like are difficult to directly play roles. Therefore, effective pretreatment is necessary before the lignocellulose resources are degraded and converted by microorganisms and enzymes to realize the industrialization of the refining technology of the lignocellulose resources. The pretreatment related research of lignocellulose has been reported in a large number, especially the fuel ethanol production process mainly using cellulose has entered the industrialization stage, and the market response is good. As a key step in the biorefinery of lignocellulose, the pretreatment process determines the feasibility of the overall refining process. The ideal pretreatment process not only aims to solve the problem of effective separation of lignin, hemicellulose and cellulose, avoid the generation of inhibitors with inhibition effect on enzymolysis and fermentation in the pretreatment process, but also improves the sugar yield as much as possible and protects the treatment process as much as possible. The high cost of pretreatment is a major limitation in lignocellulose refining from an economic point of view. The pretreatment process with high economic benefit is a precondition for realizing green and efficient refining of the whole lignocellulose.
The traditional physical and chemical pretreatment methods have the problems of high energy consumption, high cost, difficult pretreatment of waste liquid and the like; the biological pretreatment mode has the problems of long period, low efficiency and the like, and the industrial application is restricted. Most pretreatment processes are to increase the contact area of cellulase and cellulose in the enzymolysis process by removing hemicellulose and lignin in the raw materials, so as to achieve the aim of improving the enzymolysis efficiency. At present, the fuel ethanol production process taking lignocellulose resources such as corn straw and the like as raw materials mostly adopts steam explosion treatment, and the problems of low utilization rate of the whole components and difficult treatment of waste liquid are unavoidable, wherein lignin components are not well utilized. How to realize the full resource utilization of the whole lignocellulose has great significance for improving the economic benefit of the utilization of the whole lignocellulose resources.
In recent years, the technology of preferentially extracting and utilizing lignin is a hot spot for research, and mainly comprises methods such as an ionic liquid method, an organic solvent method, a DES extraction system and the like. Although the methods have less damage to lignin structures, the obtained lignin product has higher additive utilization value and improves the utilization level of lignin. However, no report on industrialization of the biorefinery process using lignin preferential extraction is currently known, and the report is limited to laboratory scale and mainly uses research mechanism. The main reasons are that the processes have the problems of high economic cost, low solvent recovery rate, harsh reaction conditions and the like, and the processes lack of matched recycling utilization processes, so that the industrial application of the processes is limited. How to solve the problem that the recycling or high-value utilization of the extraction solvent can greatly improve the green and high-efficiency utilization level of lignocellulose. Therefore, the proposal of a lignocellulose pretreatment scheme for lignin preferential extraction by a fermentable solvent has great significance for realizing the improvement of the green high-efficiency biorefinery level of lignocellulose.
Disclosure of Invention
In view of the foregoing problems with the prior art, the present application is directed to a green, efficient refining system and method for pretreating lignocellulose based on a fermentable solvent. Experiments prove that the lignocellulose refining method provided by the application not only realizes the high-value utilization of lignin, hemicellulose and cellulose, but also can overcome the defects of difficult recovery and treatment of extraction solvent and high cost, thereby having great value in large-scale industrial production and practical application.
To this end, a first aspect of the present application provides a fermentable lignocellulose pretreatment agent comprising a carbon source, an inorganic acid and betaine, wherein the carbon source comprises one or more of glucose, xylose, arabinose and glycerol, preferably glucose and/or glycerol; the inorganic acid comprises one or more of sulfuric acid, hydrochloric acid, phosphoric acid and nitric acid.
In some specific embodiments of the present application, the pretreatment agent comprises, based on the total mass of the pretreatment agent:
50% -80% of carbon source;
0.5% -5% of inorganic acid;
10% -40% of betaine;
0% -5% of water;
preferably, the molar ratio of the carbon source to betaine in the pretreatment agent is 2:1.
In some particularly preferred embodiments of the present invention, the pretreatment agent is composed of glycerin, betaine, an inorganic acid and water, wherein the molar ratio of glycerin to betaine is 2:1, and the amount of the inorganic acid is 1% based on the total mass of the pretreatment agent, and wherein the inorganic acid is phosphoric acid or sulfuric acid.
The invention provides a lignocellulose green high-efficiency refining system, which comprises a material mixing tank, a high-pressure reaction kettle, a solid residue separation device, a lignin raffinate fermentation tank, an I fermentation product separation device and an I fermentation product storage tank which are connected in sequence;
the lignin extracting solution outlet of the solid residue separation device is connected with the inlet of the lignin separation device;
the lignin raffinate outlet of the lignin separation device is connected with the inlet of the lignin raffinate fermentation tank;
the lignin refining system also comprises a lignin storage tank, wherein the inlet of the lignin storage tank is connected with the lignin outlet of the lignin separation device;
the system also comprises a lignin extracting solution circulating pipeline, one end of the lignin extracting solution circulating pipeline is connected with the outlet of the solid-liquid residue separating device, and the other end of the lignin extracting solution circulating pipeline is connected with the inlet of the material mixing tank, so that the material mixing tank, the high-pressure reaction kettle, the solid residue separating device and the lignin extracting solution circulating pipeline form a lignin extracting solution circulating loop.
The system further comprises a moisture evaporation device, wherein the inlet of the moisture evaporation device is connected with the lignin raffinate outlet of the lignin separation device, and the non-moisture material outlet of the moisture evaporation device is connected with the inlet of the material mixing tank, so that the material mixing tank, the high-pressure reaction kettle, the solid residue separation device, the lignin separation device and the moisture evaporation device form a lignin raffinate circulation loop.
According to some embodiments of the invention, the system further comprises an enzymolysis tank, an enzymolysis liquid fermentation tank, a second fermentation product separation device and a second fermentation product storage tank connected to the solid residue outlet of the solid residue separation device.
According to other embodiments of the present invention, the system further comprises a mixed liquor fermentation tank, a third fermentation product separation device and a third fermentation product storage tank which are connected in sequence, wherein the inlet of the mixed liquor fermentation tank is respectively connected with the lignin raffinate outlet of the lignin separation device and the outlet of the enzymolysis tank; the first fermentation product storage tank is the same as or different from the third fermentation product storage tank.
A third aspect of the present invention provides a method for green efficient refining of lignocellulose using the green efficient refining system of the second aspect of the present invention, comprising:
Step A, mixing a pretreatment agent and a biomass raw material in a material mixing tank, and then sending the mixture into a high-temperature reaction kettle for high-temperature pretreatment to obtain a high-temperature pretreatment product;
step B, separating and treating the high-temperature pretreatment product through a solid residue separating device to obtain lignin extract and solid residue;
step C, separating and treating part of lignin extracting solution through a lignin separation device to obtain lignin raffinate and lignin;
step D, fermenting a part of lignin raffinate through a lignin raffinate fermentation tank to obtain a lignin raffinate fermentation culture product, and separating the lignin raffinate fermentation culture product through a first fermentation product separation device to obtain a first fermentation product;
wherein the pretreatment agent is the fermentable lignocellulose pretreatment agent according to the first aspect of the present invention.
In some embodiments of the present invention, the lignocellulose green efficient refining method further includes step E, sending the lignin extracting solution obtained in step B back to the material mixing tank as a solvent for the next pretreatment, and repeating step a and step B until the pretreatment effect is reduced to below 70% of the lignin initial removal rate;
In other embodiments of the present invention, the green efficient refining method for lignocellulose further includes step F, the lignin raffinate obtained in step C is sent back to the material mixing tank as a solvent for the next pretreatment after being dehydrated by the water evaporation device, and steps a, B and C are repeated until the lignin removal rate is reduced to less than 70% of the lignin initial removal rate.
According to some embodiments of the invention, the green efficient refining method of lignocellulose further comprises step G, performing enzymolysis on the solid residues through an enzymolysis tank, performing sugar liquor fermentation treatment on the obtained enzymolysis liquor through an enzymolysis liquor fermentation tank, and then performing separation treatment on the sugar liquor fermentation product through a second fermentation product separation device to obtain a second fermentation product.
According to other embodiments of the present invention, the green efficient refining method for lignocellulose further includes step H, fermenting a mixture of a part of lignin raffinate and enzymolysis liquid by a mixed fermentation tank, and separating a mixture fermentation product by a third fermentation product separating device to obtain a third fermentation product.
The invention has the following beneficial effects:
(1) The invention provides a full-component green efficient refining process of lignocellulose based on fermentable solvent pretreatment, which combines the current utilization situation of lignocellulose: the selected pretreatment solvents can be used as carbon sources of microorganisms to ferment and produce related products, and the pretreatment cost is reduced by recycling the pretreatment solvents.
(2) The invention realizes the high-value utilization of 3 components in lignocellulose to the maximum extent, wherein the lignin structure obtained by pretreatment and extraction is protected to the maximum extent, and the product quality and the added value are high. The pretreatment process dissolves out most of the hemicellulose as a component available to microorganisms to enter the fermentation to increase the content of the target product. The pretreated residues can be better subjected to enzymolysis due to the removal of lignin and hemicellulose, so that the cellulose components can be further utilized as sugar resources.
(3) The solvent in the invention can be recycled for a plurality of times, can realize the recycling of resources, and meets the current green manufacturing requirement.
(4) The process provided by the invention has wide applicability, is obtained by amplifying and optimizing the process on the basis of simulation experiment conditions, and is suitable for popularization and utilization.
(5) The solvent related by the invention is a raw material which is green and environment-friendly, has low price and is easy to obtain, the process is nontoxic and harmless, and substances harmful to the environment such as heavy metals, waste liquid and the like are not produced.
In general, the process disclosed by the invention comprises a full resource utilization scheme of lignocellulose, realizes green and efficient refining of all components of lignin, hemicellulose and cellulose, provides a new idea of green and efficient utilization of lignocellulose resources, effectively reduces the cost of the process, and expands the processing industry chain of the lignocellulose resources. Is helpful to promote the comprehensive utilization level of renewable lignocellulose resources in China.
Drawings
In order that the invention may be readily understood, the invention will be described in detail below with reference to the accompanying drawings.
Fig. 1 is a schematic diagram of a full component green high efficiency refining process flow of lignocellulose.
Fig. 2 is a schematic diagram of a green high-efficiency refining system for lignocellulose.
Fig. 3 is a schematic diagram of a green high-efficiency refining system for lignocellulose.
The reference numerals in fig. 2 and 3 are explained as follows: 1 a material mixing tank; 2, a high-pressure reaction kettle; 3 a solid-liquid residue separating device; 4 lignin extracting solution circulating pipeline; 10 lignin extract three-way valve or lignin extract storage tank; 11 lignin separation means; a lignin raffinate four-way valve or lignin raffinate storage tank; 13 lignin raffinate fermentor; 14 first fermentation product separation means; 15 th fermentation product storage tank; a 16 lignin storage tank; 17 a moisture evaporation device; 20 a solid-liquid residue storage tank; 21 enzymolysis tank; 22 an enzymolysis liquid three-way valve or an enzymolysis liquid storage tank; 23 an enzymolysis liquid fermentation tank; a second fermentation product separation device 24; a 25 th fermentation product tank; 32 a mixed liquor three-way valve or a mixed liquor tank; 33 a mixed liquor fermentation tank; 34 a III fermentation product separation device; and 35 III fermentation product storage tank.
Detailed Description
In order that the invention may be readily understood, a more particular description of the invention will be rendered by reference to specific embodiments that are illustrated in the appended drawings. Before the present invention is described in detail, it is to be understood that this invention is not limited to particular embodiments described. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.
Where a range of values is provided, it is understood that each intervening value, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and are also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.
Unless defined otherwise, all terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, the preferred methods and materials are now described.
I terminology
The term "fermentable lignocellulose pretreatment agent" in the present invention is a mixed solvent formed by mixing two or more components, wherein the mixed solvent comprises at least one carbon source capable of being utilized by microorganisms and other components, and the other components do not cause obvious inhibition to the growth of the microorganisms. The pretreatment agent can be used to perform pretreatment of lignocellulose to achieve efficient separation of lignin, cellulose and hemicellulose.
The term "lignocellulose green high-efficiency refining method" in the invention specifically refers to a method for effectively separating lignin, cellulose and hemicellulose after pretreatment of lignocellulose, respectively obtaining products with high added value after processing, effectively utilizing all components of lignocellulose, and also obtaining products with high added value after processing solvents and the like used in the pretreatment process, wherein no waste is discharged in the whole process, and the method accords with the concept of green processing.
The term "lignocellulose full-component green efficient refining process" in the invention specifically refers to a specific means and a specific process route for realizing the "lignocellulose green efficient refining method", relates to the connection and sequence of each processing link, and comprises the steps of adding solvent circulation, water resource recycling, heat recovery and the like, and can be specifically referred to a process flow chart.
The term "fermentable lignocellulose pretreatment agent" and "fermentable pretreatment solvent" as used herein may be used interchangeably.
II. Embodiment
As described above, the existing technologies for producing lignocellulose and producing biological products are always unsatisfactory, for example, the technologies for producing lignocellulose and producing biological products cannot realize the full resource utilization of the whole lignocellulose, and the technologies for producing lignocellulose and producing biological products have the problems of high economic cost, low solvent recovery rate, harsh reaction conditions and the like, and lack of matched resource utilization technologies, so that the industrial application of lignocellulose is limited. In order to solve the above problems in the prior art, the present inventors have conducted extensive and intensive studies on technologies for producing lignocellulose in biofuel and producing biological products, and have studied and devised a green and efficient refining method of lignocellulose, which is capable of better achieving full-resource utilization of the whole lignocellulose in the production of lignocellulose in biofuel and in the production of biological products, wherein a schematic process flow diagram is shown in fig. 1. Experiments prove that the lignocellulose refining method provided by the application not only realizes the high-value utilization of lignin, hemicellulose and cellulose, but also can overcome the defects of difficult recovery and treatment of extraction solvent and high cost, thereby having great value in large-scale industrial production and practical application.
To achieve the above technical solution, according to a first aspect of the present invention, there is provided a fermentable lignocellulose pretreatment agent which can be directly used for fermentation as a lignocellulose pretreatment agent, and which comprises a carbon source, an inorganic acid and betaine, wherein the carbon source comprises one or more of glucose, xylose, arabinose and glycerin, preferably glucose and/or glycerin, more preferably glycerin; the inorganic acid comprises one or more of sulfuric acid, hydrochloric acid, phosphoric acid and nitric acid, preferably phosphoric acid.
In some specific embodiments of the present invention, the pretreatment agent comprises, based on the total mass of the pretreatment agent:
50% -80% of carbon source;
0.5% -5% of inorganic acid;
10% -40% of betaine;
0% -5% of water;
preferably, the molar ratio of the carbon source to betaine in the pretreatment agent is 2:1.
In some particularly preferred embodiments of the present invention, the pretreatment agent is composed of glycerin, betaine, an inorganic acid and water, wherein the molar ratio of glycerin to betaine is 2:1, and the amount of the inorganic acid is 1% based on the total mass of the pretreatment agent, and wherein the inorganic acid is phosphoric acid or sulfuric acid.
In a second aspect, the invention provides a lignocellulose green high-efficiency refining system, and the process flow diagram of the system is shown in fig. 2. As can be seen from fig. 2, the lignocellulose green high-efficiency refining system comprises a material mixing tank 1, a high-pressure reaction kettle 2, a solid residue separating device 3, a lignin separating device 11, a lignin raffinate fermenter 13, an I fermentation product separating device 14 and an I fermentation product storage tank 15 which are connected in sequence;
The lignocellulose extracting solution outlet of the solid residue separating device 3 is connected with the inlet of the lignin separating device 11;
the lignin raffinate outlet of the lignin separation unit 11 is connected with the inlet of the lignin raffinate fermentation tank 13;
the lignin refining system further comprises a lignin storage tank 16, the inlet of which is connected with the lignin outlet of the lignin separation unit 11;
the system also comprises a lignin extracting solution circulating pipeline, one end of the lignin extracting solution circulating pipeline is connected with the outlet of the solid-liquid residue separating device 11, and the other end of the lignin extracting solution circulating pipeline is connected with the inlet of the material mixing tank 1, so that the material mixing tank 1, the high-pressure reaction kettle 2, the solid residue separating device 3 and the lignin extracting solution circulating pipeline 4 form a lignin extracting solution circulating loop.
The system further comprises a moisture evaporation device 17, the inlet of which is connected with the lignin raffinate outlet of the lignin separation device 11, and the non-moisture material outlet of which is connected with the inlet of the material mixing tank 1, whereby the material mixing tank 1, the autoclave 2, the solid residue separation device 3, the lignin separation device 11, and the moisture evaporation device 17 form a lignin raffinate circulation loop.
According to some embodiments of the invention, the system further comprises an enzymolysis tank 21, an enzymolysis liquid fermentation tank 23, a second fermentation product separation device 24 and a second fermentation product tank 25 connected to the solid residue outlet of the solid residue separation device 3.
According to further embodiments of the present invention, the system further comprises a mixed liquor fermentation tank 33, a III fermentation product separation device 34 and a III fermentation product storage tank 35, which are connected in sequence, wherein the inlet of the mixed liquor fermentation tank 33 is connected to the lignin raffinate outlet of the lignin separation device 11 and the outlet of the enzymatic hydrolysis tank 22, respectively.
Preferably, the three-way valve 10, the four-way valve 12, the three-way valve 32 and the three-way valve 22 in the process flow shown in fig. 2 are respectively set as the lignin extracting solution storage tank 10, the lignin raffinate storage tank 12, the mixed liquid tank 32 and the enzymolysis solution storage tank 22, and a solid-liquid residue storage tank is additionally arranged between the solid-liquid residue separation device 3 and the enzymolysis tank 21 and is used as a buffer storage tank for corresponding materials so as to improve the operation flexibility and the flexibility, as shown in fig. 3.
A third aspect of the present invention provides a method for green and efficient refining of lignocellulose using the green and efficient refining system of lignocellulose according to the second aspect of the present invention, which can be understood as a process for green and efficient refining of lignocellulose using the green and efficient refining system of lignocellulose according to the second aspect of the present invention, as shown in fig. 1, comprising:
step A, mixing a pretreatment agent and a biomass raw material (namely lignocellulose) in a material mixing tank 1, and then sending the mixture into a high-temperature reaction kettle 2 for high-temperature pretreatment to obtain a high-temperature pretreatment product;
Step B, separating and treating the high-temperature pretreatment product through a solid residue separating device 3 to obtain lignin extract and solid residue;
step C, separating and treating part of lignin extracting solution through a lignin separation device 11 to obtain lignin raffinate and lignin;
step D, fermenting a part of lignin raffinate through a lignin raffinate fermentation tank 13 to obtain a lignin raffinate fermentation culture product, and separating the lignin raffinate fermentation culture product through a fermentation product separation device 14 to obtain a fermentation product I;
wherein the pretreatment agent is the fermentable lignocellulose pretreatment agent according to the first aspect of the present invention.
In some embodiments of the present invention, the lignocellulose green efficient refining method further includes step E, sending the lignin extracting solution obtained in step B back to the material mixing tank 1 as a solvent for the next pretreatment, and repeating step a and step B until the pretreatment effect is reduced to less than 70% of the lignin initial removal rate;
in other embodiments of the present invention, the green efficient refining method for lignocellulose further includes step F, the lignin raffinate obtained in step C is sent back to the material mixing tank 1 as a solvent for the next pretreatment after being dehydrated by the moisture evaporation device 17, and steps a, B and C are repeated until the pretreatment effect is reduced to less than 70% of the initial lignin removal rate.
According to some embodiments of the present invention, the green and efficient refining method for lignocellulose further comprises step G, performing enzymolysis on the solid residues through an enzymolysis tank 21, performing sugar liquor fermentation treatment on the obtained enzymolysis liquor through an enzymolysis liquor fermentation tank 23, and then performing separation treatment on the sugar liquor fermentation product through a second fermentation product separation device 24 to obtain a second fermentation product.
According to other embodiments of the present invention, the green and efficient refining method for lignocellulose further includes step H, fermenting a mixture of a part of lignin raffinate and enzymatic hydrolysate by using a mixing fermenter 33, and separating the mixture fermentation product by using a third fermentation product separating device 34 to obtain a third fermentation product.
It is to be readily understood that the above process is realized based on the fermentable lignocellulose pretreatment agent according to the first aspect of the present invention and the lignocellulose green high-efficiency refining system according to the second aspect of the present invention, and thus the lignocellulose green high-efficiency refining system and method provided by the present invention can be understood as a lignocellulose green high-efficiency refining system and method based on the fermentable pretreatment agent.
In some specific embodiments of the present invention, the lignocellulosic green efficient refining method specifically comprises the steps of:
1) Selection of raw materials
The production process provided by the invention is applicable to most lignocellulose of plant origin, including but not limited to the following categories: energy grass (reed, switchgrass, pennisetum), wood (poplar, willow, eucalyptus), agricultural waste (corn stover, wheat straw, sorghum straw), and the like.
2) Crushing of raw materials
The obtained raw materials are mechanically crushed, and different crushing modes are selected according to different raw materials in the crushing process, including multistage crushing.
3) Screening and dust removal
The purpose of screening and dedusting is mainly to remove fine sand, dust and the like entrained in raw materials, so that abrasion of equipment and consumption of chemical in the process in the subsequent steps are reduced. The dedusting means include, but are not limited to, sieving, air separation, and other means known to the scientific researchers.
4) Preparation of fermentable pretreatment solvent
The pretreatment solvents involved in the invention are all carbon sources, nitrogen sources, inorganic salts and the like which can be utilized by microorganisms, and comprise common carbon sources well known in the art, such as: glucose, xylose, arabinose, glycerol, etc.; other substances which can be used as auxiliary materials and are not obviously inhibited to microorganisms after being added into a culture medium are verified, such as: betaines.
One or more of the substances are selected to be optionally combined for preparing the pretreatment solvent, and water is added or not added according to actual requirements.
5) High temperature pretreatment
Adding lignocellulose raw materials into the solvent prepared in the step 4) according to a certain proportion, uniformly mixing, and adding into a high-temperature high-pressure reaction kettle for high-temperature pretreatment.
In this step, different solid to liquid ratios are selected according to different raw materials, for example, the solid to liquid ratio of the lignocellulose raw material to the solvent may be 1 (1-20).
6) Solid-liquid separation
Filtering or centrifuging the reaction product obtained in the step 5) to obtain lignin extract and cellulose filter residues.
7) Washing of cellulose filter residue
And (3) washing the cellulose filter residue obtained in the step 6).
8) Lignin extraction
Diluting the lignin extract obtained in the step 6) by adding water, and regulating the pH value to 3-7. Lignin is separated out from the solution after standing.
9) Lignin separation
Subjecting the cloudy solution obtained in step 8) to solid-liquid separation including, but not limited to, centrifugation or filtration to obtain lignin solids and lignin raffinate, respectively. The obtained lignin solid is dried and subjected to structural analysis, and can be used as a raw material for other processes, including but not limited to being used as an additive of a 3D printing material, preparing other lignin-based composite materials and the like.
10 Pretreatment of the extract
And selecting whether to perform decolorization treatment according to the quality of lignin raffinate after extracting lignin, wherein the using amount of activated carbon is 0.1%.
11 Preparation of fermentation Medium
The diluted solution of the lignin raffinate obtained by the separation after the decolorization in the step 10) contains a large amount of substances which can be utilized by microorganisms, and other nutrient substances which are necessary for the growth of the microorganisms are added into the diluted solution of the lignin raffinate obtained in the step 10) according to the requirements of target products. And adjusting the pH value of the extract dilution according to different fermentation processes, and then sterilizing. The dilution liquid is diluted according to the concentration of the conventional culture medium, and the specific dilution ratio is determined after optimization according to the tolerance and the like of the actual cultured microorganisms. Other nutrients necessary for the growth of the microorganism are conventional nutrients required in the art for the cultivation of the relevant microorganism.
12 Fermentation of the extract
And step 10), inoculating microorganisms to the sterilized culture medium, and fermenting according to different target products and process control. The target products include, but are not limited to, microbial oils, citric acid, erythritol, and the like.
13 Use of cellulose components
And 7) carrying out enzymolysis on the cellulose filter residue obtained in the step 7) to obtain a glucose-based sugar-containing enzymolysis liquid, and carrying out the next utilization, wherein the utilization modes comprise, but are not limited to, acetone fermentation, ethanol fermentation, butanol fermentation and the like.
The enzyme used for the enzymatic hydrolysis in the present invention is not particularly limited, and enzymatic hydrolysis may be performed using cellulase which is conventional in the art, for example, cellulase cellular CTec3 (Novozymes).
14 Repeatedly using fermentable pretreatment solvent
The prepared fermentable pretreatment agent can be recycled, after lignin is extracted, solid cellulose residues are removed, and the residual liquid part can be further used as the pretreatment agent for lignin extraction. For this reason, the above steps are single pretreatment steps, and in actual practice, after step 6), the lignin extract obtained in step 6) may be reused as a solvent for the next pretreatment until the lignin removal rate is reduced to 70% or less of the initial removal rate.
The lignin raffinate separated in the step 9) may be subjected to water evaporation (water evaporation device 17) and then reused as a solvent for the next pretreatment until the removal rate is reduced to 70% or less of the initial removal rate.
15 Mixed fermentation of extracting solution and enzymolysis solution
The process involves fermentation of two parts of material. One part is lignin raffinate fermentation, and the other part is cellulose residue fermentation after enzymolysis. The two parts can be fermented respectively, or the two parts of materials can be mixed and then fermented. Mixing the lignin raffinate obtained by the decolorization and separation obtained in the step 13) and the step 10) according to a certain proportion, and then adding a necessary nitrogen source for fermentation. The nitrogen source is a nitrogen source used in fermentation processes conventional in the art.
In the preferred technical scheme of the invention, the crushing of the raw materials in the step 2) is divided into multiple stages of crushing, and the raw materials are preferably sieved by a sieve with 40-80 meshes. In a further preferred embodiment, the final size of the particles is less than 0.2 mm. Cutting the materials into 1-5cm by primary crushing, wherein the size and diameter of the raw materials after secondary crushing are less than 1 millimeter, and finally the particle size of the raw materials after further crushing is less than 0.2 millimeter.
Preferably, the fermentable pretreatment solvent in the step 4) is determined to be one or more of glucose, glycerol and betaine, and inorganic acid (sulfuric acid, hydrochloric acid, phosphoric acid, nitric acid and the like) is added or not added by comprehensively considering the dissolution rate of lignin and the cellulose retention rate. Further preferred combinations are glycerol, betaine, phosphoric acid, and optionally water. Preferably, the component proportions of the fermentable solvent system are: 50-80% of glycerol, 10-40% of betaine and 0-5% of water. Further preferred components are as follows: 55-65% of glycerol, 30-40% of betaine and 0-5% of water. More preferably, the composition of the fermentable solvent system is in molar ratio: glycerol: betaine = 2:1 (1% phosphoric acid). More preferably, the temperature of the high temperature treatment of step 5) is 100-300 ℃, and even more preferably the extraction temperature is 120-160 ℃. The high temperature treatment time in the step 5) is 10min-10h, and the preferred extraction time is 2-4 h.
The solid-liquid separation mode of the step 6) is adjusted according to the experimental scale, and the small-scale extraction process in the laboratory can adopt modes of centrifugation, suction filtration and the like. The pilot and scale-up processes may be disc centrifugation, drum filtration, doctor blade centrifugation, plate and frame filtration, or may be a combination of several separation methods or other solid-liquid separation methods known in the art.
Preferably, the waste water after washing in the step 7) can be used as the extracting solution obtained in the step 8) to dilute, thereby achieving the purpose of environmental protection.
Step 8) diluting the extract obtained in step 6), preferably by 3-10 times, and more preferably by 4-6 times. At the same time, the pH of the extract is preferably adjusted to 4-5.
The lignin separation method in the step 9) includes, but is not limited to, centrifugation, suction filtration, plate-frame filtration and the like. Preferably, the separation mode of the lignin of the small-scale process in the laboratory is centrifugation, and the separation mode of the lignin of the pilot scale and the large-scale process is plate-frame filtration.
The lignin drying method in step 9) includes, but is not limited to, direct drying, vacuum freeze drying, fluidized bed drying, air drying, microwave drying, etc. The preferred drying means is vacuum freeze drying.
The pretreatment mode in the step 10) comprises centrifugation, membrane filtration and the like. Preferred membrane filtration means include ceramic membranes, nanofiltration, microfiltration, etc.
Step 11) and step 12) are carried out by adding reasonable substances according to the required target product, in a preferred scheme, the main components of the extraction solvent are glycerol and phosphoric acid, the selected microorganism can ferment by taking glycerol as a carbon source, the preferred strain is oleaginous yeast (rhodochrogullutinis), and the products are microbial grease and carotenoid.
Preferably, the mixed fermentation of the extracting solution and the enzymolysis solution in the step 15) is carried out in combination with the step 14), and when the fermentable solvent is used for pretreatment for a plurality of times, the enzymolysis solution and the fermentable solvent are compounded, and then the grease fermentation is carried out.
In the scheme, researchers can adjust the composition and proportion of the extraction solvent according to different requirements, and select proper strains for fermentation according to different extraction solvents to obtain different products.
And 9) drying the lignin to obtain lignin of different types, and selecting different utilization schemes according to different lignin structures.
The cellulose residue after lignin extraction in step 13) may be treated according to the currently established enzymatic hydrolysis process to obtain a glucose-based sugar-containing liquid, which is then subjected to a further fermentation process, including but not limited to acetone, butanol, ethanol fermentation.
A specific process flow diagram is shown in FIG. 1, wherein the first fermentation product comprises but is not limited to grease, the first fermentation product comprises but is not limited to acetone, butanol, ethanol, and the third fermentation product comprises but is not limited to grease.
The above preferred embodiments are the optimization scheme proposed by the inventor aiming at the lignocellulose full-component green efficient refining process, and the above preferred process conditions are proposed by performing process amplification and optimization through simulation experiments. According to the technical scheme, the whole process does not involve toxic and harmful solvents according to the chemical safety requirements, and the process is green and environment-friendly and accords with corresponding laws and regulations.
III. Examples
In order that the invention may be more readily understood, a more particular description of the invention will be rendered by reference to specific embodiments that are illustrated in the appended drawings and are therefore not to be considered limiting of its scope. The starting materials or components used in the present invention may be obtained commercially or by conventional methods unless otherwise specified.
Enzymatic hydrolysis was performed using cellulase cellular CTec3 (Novozymes).
The strain used in the embodiment of the invention for producing the microbial oil is rhodotorula glutinis (CGMCC No. 2258), and is stored in a refrigerator at the temperature of minus 80 ℃ in a glycerol pipe, and is activated once on a solid flat plate when in use. The yeast for producing ethanol is Saccharomyces cerevisiae active dry yeast, purchased from Angel yeast.
Oil fermentation medium: a proper amount of lignin raffinate, 2g/L urea, 1.5g/L Y yeast powder and 2g/L dipotassium hydrogen phosphate.
Ethanol fermentation medium: proper amount of enzymolysis liquid, 0.5g/L diammonium hydrogen phosphate, 0.5g/L dipotassium hydrogen phosphate, 0.5g/L peptone and 0.5g/L yeast powder.
Preparation of raw materials: taking the dried raw materials, cutting the raw materials into blocks, mechanically crushing the blocks, and sieving the crushed blocks with a 40-mesh sieve to obtain the experimental raw materials. The large-scale production adopts special crushing equipment and screening equipment.
Preparation of pretreatment solvent system: mixing different glycerin, inorganic acid, betaine, etc. in certain molar ratio, heating to 60-80deg.C, and stirring for 120min to obtain uniform colorless transparent pretreatment agent.
The lignin removal rate was calculated as follows:
the raw materials mainly used in the following examples are poplar, reed and corn stalks.
Example 1: determination of main components of poplar, reed and corn straw
The ratio of cellulose, hemicellulose and lignin in the raw material is determined by a two-step sulfuric acid method. Specifically, 0.2g of the ground dry sample was added to 3mL of 72% H 2 SO 4 To the solution, 112mL of deionized water was added after stirring for 120min, and the mixture was hydrolyzed at 121℃for 60min. And (3) carrying out suction filtration to obtain acid-insoluble lignin, wherein the acid-insoluble lignin is measured by an ultraviolet-visible light spectrometer (wavelength-205 nm). The hydrolyzed saccharide fraction was analyzed by HPLC (Thermo Scientific, U3000). The results are shown in Table 1:
TABLE 1
Cellulose (%) Hemicellulose (%) Lignin (%) Other (%)
Poplar 48 25 21 6
Reed 44 27 20 9
Corn stalk 42.5 23.5 19 15
Example 2:
500g of pretreatment solvent was prepared by mixing glycerol and betaine in a molar ratio of 2:1, wherein the glycerol was 60.4%, the betaine was 38.6% and the phosphoric acid was 1%. 50g of poplar is weighed and reacted for 2 hours at 140 ℃, and 24.9g of cellulose residue is obtained after suction filtration and drying. The liquid part is added with water to 3L, and 6.1g of lignin is obtained after precipitation treatment. The cellulose content in the residue was measured to be 79.1%, hemicellulose content was measured to be 2.8%, and lignin content was measured to be 7.4%. Wherein the pretreatment solvent is measured to have a glycerol concentration of 98.9g/L and a betaine concentration of 60.3g/L. And then adding nutrient elements and inorganic salts into the pretreatment solvent according to the formula of the culture medium for oil fermentation. After sterilization, the grease fermentation is carried out, and after 360 hours, the dry weight of the thalli is measured to be 35.1g/L, and the grease concentration is measured to be 10.1g/L (product I). Adding phosphate buffer solution into cellulose residue according to the concentration of 6% of solid content, and obtaining enzymolysis solution with the glucose concentration of 46.4g/L after enzymolysis. And (3) adding nutrient substances into the enzymolysis liquid after centrifugation according to an ethanol fermentation medium formula, and carrying out ethanol fermentation, wherein the concentration of ethanol in the fermentation liquid is 18.6g/L (product II) after the final fermentation.
Example 3:
500g of pretreatment solvent is prepared by glycerol and betaine according to the mol ratio of 2:1, wherein 60.4 percent of glycerol and 38.6 percent of betaine are mixed uniformly, and then 1 percent of sulfuric acid is added. 50g of corn straw is weighed and reacted for 2 hours at 140 ℃, and 23.2g of cellulose residue is obtained after suction filtration and drying. The liquid part is added with water to 3L, and 4.7g of lignin is obtained after precipitation treatment. The cellulose content in the residue was found to be 76.2%, hemicellulose content was found to be 2.8% and lignin content was found to be 8.2%. Wherein the pretreatment solvent is measured to have a glycerol concentration of 100.1g/L and a betaine concentration of 64.8g/L. Then adding nutrient elements and inorganic salts (without adding phosphate) into the pretreatment solvent according to the formula of the culture medium for oil fermentation. After sterilization, the grease fermentation is carried out, after 360 hours, the dry weight of the thalli is measured to be 37.1g/L, and the grease concentration is measured to be 11.7g/L (product I). Adding phosphate buffer solution into cellulose residue according to the concentration of 6% of solid content, and obtaining enzymolysis solution with glucose concentration of 44.9g/L after enzymolysis. And (3) adding nutrient substances into the enzymolysis liquid after centrifugation according to the formula of the ethanol fermentation medium, and carrying out ethanol fermentation, wherein the concentration of ethanol in the fermentation liquid is 17.6g/L (product II) after the final fermentation.
Example 4:
500g of pretreatment solvent was prepared with glycerin and betaine in a molar ratio of 2:1, wherein glycerin was 59.8%, betaine was 38.2%, and phosphoric acid was 2%. 50g of poplar is weighed and reacted for 2 hours at 140 ℃, and 25.1g of cellulose residue is obtained after the solid part is washed and dried after centrifugation, and the liquid part is weighed to 400g, and the first repeated reaction is counted. Then adding 40g poplar into 400g liquid to continue to react for 2 hours at 140 ℃, centrifuging, washing and drying the solid part to obtain 21.5g cellulose residue, weighing 295g liquid part, and repeating the reaction for the second time. 29.5g poplar was then added to 295g liquid and reacted for 2 hours at 140℃and after centrifugation the solid fraction was washed and dried to give 16.3g of cellulose residue, the liquid fraction weighed 218g, counting a third repeat reaction. Then adding 22g poplar into 218g liquid to continue to react for 2 hours at 140 ℃, centrifuging, washing and drying the solid part to obtain 16.6g cellulose residue, and weighing 135g liquid part for the fourth repeated reaction. The liquid fraction of the fourth repeated reaction was fixed to 500mL and precipitated to obtain 11.3g of lignin in total.
Then, the lignin-recovered liquid was subjected to moisture removal by a rotary evaporator to obtain 122g of an anhydrous liquid. Adding 12g poplar into 122g liquid according to a proportion, continuously reacting for 2 hours at 140 ℃, centrifuging, washing and drying a solid part to obtain 6.7g cellulose residue, weighing 75g liquid part, and calculating as a fifth repeated reaction. Then adding 7.5g poplar into 75g liquid to continue to react for 2 hours at 140 ℃, and obtaining 3.9g cellulose residue after centrifugation and washing and drying of solid part, wherein the liquid part is weighed to be 50g, and the sixth repeated reaction is counted. Then adding 5g poplar into 50g liquid to continue to react for 2 hours at 140 ℃, and obtaining 4.1g cellulose residue after centrifugation and washing and drying of solid part, wherein the liquid part is weighed to be 30g, and the seventh repeated reaction is calculated. The liquid portion of the seventh repeat reaction was sized to 150mL and precipitated to yield 1.96g lignin. Then the different components in the cellulose residues obtained from the first to fourth times were measured respectively, see Table 2
TABLE 2 cellulose residue Components at different repetition times
Example 5:
73.5g of cellulose residues obtained in the first, second, third, fifth and sixth steps in the example 4 are respectively collected, and then enzymolysis is carried out according to the solid content of 10%, so that the glucose concentration of the enzymolysis liquid is 74.3g/L, and the xylose concentration is 5.8g/L. 150mL of the enzymatic hydrolysate was mixed with 150mL of the liquid obtained in the seventh step of example 4 to obtain a mixed solution containing the raffinate and the enzymatic hydrolysate. The concentration of glucose in the mixed solution is 36.2g/L, the concentration of xylose is 2.6g/L, and the concentration of glycerol is 57.2g/L. After sterilization, the oil fermentation was carried out for 400 hours, and after the fermentation was completed, the dry weight of the cells was measured to be 39.1g/L, and the oil concentration was measured to be 12.1g/L (product III).
It should be noted that the above-described embodiments are only for explaining the present invention and do not constitute any limitation of the present invention. The invention has been described with reference to exemplary embodiments, but it is understood that the words which have been used are words of description and illustration, rather than words of limitation. Modifications may be made to the invention as defined in the appended claims, and the invention may be modified without departing from the scope and spirit of the invention. Although the invention is described herein with reference to particular means, materials and embodiments, the invention is not intended to be limited to the particulars disclosed herein, as the invention extends to all other means and applications which perform the same function.

Claims (7)

1. A pretreatment agent for fermenting lignocellulose, which consists of glycerin, betaine and inorganic acid; the inorganic acid is sulfuric acid or phosphoric acid, the mol ratio of glycerin to betaine is 2:1, and the dosage of the inorganic acid is 1% -2% based on the total mass of the pretreatment agent, wherein the glycerin is 59.8% -60.4%, the betaine is 38.2% -38.6%.
2. A method of lignocellulosic refining using a lignocellulosic refining system, comprising:
step A, mixing a pretreatment agent and a biomass raw material in a material mixing tank, and then sending the mixture into a high-temperature reaction kettle for high-temperature pretreatment to obtain a high-temperature pretreatment product;
step B, separating and treating the high-temperature pretreatment product through a solid residue separating device to obtain lignin extract and solid residue;
step C, separating and treating part of lignin extracting solution through a lignin separation device to obtain lignin raffinate and lignin;
step D, fermenting a part of lignin raffinate through a lignin raffinate fermentation tank to obtain a lignin raffinate fermentation culture product, and separating the lignin raffinate fermentation culture product through a first fermentation product separation device to obtain a first fermentation product;
Wherein the pretreatment agent is the pretreatment agent for fermenting lignocellulose according to claim 1;
the first fermentation product is grease;
the lignocellulose refining system comprises a material mixing tank, a high-pressure reaction kettle, a solid residue separation device, a lignin raffinate fermentation tank, a first fermentation product separation device and a first fermentation product storage tank which are connected in sequence;
the lignin extracting solution outlet of the solid residue separation device is connected with the inlet of the lignin separation device;
the lignin raffinate outlet of the lignin separation device is connected with the inlet of the lignin raffinate fermentation tank;
the lignin refining system also comprises a lignin storage tank, wherein the inlet of the lignin storage tank is connected with the lignin outlet of the lignin separation device;
the system also comprises a lignin extracting solution circulating pipeline, wherein one end of the lignin extracting solution circulating pipeline is connected with the outlet of the solid-liquid residue separating device, and the other end of the lignin extracting solution circulating pipeline is connected with the inlet of the material mixing tank, so that the material mixing tank, the high-pressure reaction kettle, the solid residue separating device and the lignin extracting solution circulating pipeline form a lignin extracting solution circulating loop;
the system also comprises a moisture evaporation device, wherein the inlet of the moisture evaporation device is connected with a lignin raffinate outlet of the lignin separation device, and the non-moisture material outlet of the moisture evaporation device is connected with an inlet of the material mixing tank, so that the material mixing tank, the high-pressure reaction kettle, the solid residue separation device, the lignin separation device and the moisture evaporation device form a lignin raffinate circulation loop;
The system also comprises an enzymolysis tank, an enzymolysis liquid fermentation tank, a second fermentation product separation device and a second fermentation product storage tank which are connected with the solid residue outlet of the solid residue separation device;
the system also comprises a mixed liquor fermentation tank, a third fermentation product separation device and a third fermentation product storage tank which are sequentially connected, wherein the inlet of the mixed liquor fermentation tank is respectively connected with the lignin raffinate outlet of the lignin separation device and the outlet of the enzymolysis tank.
3. The method according to claim 2, further comprising a step E of returning the lignin extraction solution obtained in the step B as a solvent for the subsequent pretreatment to the material mixing tank, and repeating the steps a and B until the lignin removal rate is reduced to 70% or less of the initial lignin removal rate.
4. The method according to claim 2, further comprising a step F of returning the lignin raffinate obtained in the step C to the material mixing tank as a solvent for the subsequent pretreatment after dehydration by the water evaporation device, and repeating the steps a, B and C until the pretreatment effect is reduced to less than 70% of the initial lignin removal rate.
5. A method of refining lignocellulose according to claim 3, further comprising a step F of returning the lignin raffinate obtained in the step C to the material mixing tank as a solvent for the subsequent pretreatment after dehydration by the water evaporation device, and repeating the steps a, B and C until the pretreatment effect is reduced to less than 70% of the initial lignin removal rate.
6. The method according to any one of claims 2 to 5, wherein the method further comprises step G of subjecting the solid residue to enzymolysis by an enzymolysis tank, subjecting the obtained enzymolysis liquid to a sugar liquor fermentation treatment by an enzymolysis liquor fermentation tank, and then subjecting the sugar liquor fermentation product to a separation treatment by a ii fermentation product separation device to obtain a ii fermentation product; wherein the second fermentation product comprises acetone, butanol and ethanol.
7. The method for refining lignocellulose according to claim 6, further comprising the steps of H, fermenting a part of the mixture of lignin raffinate and enzymatic hydrolysate by means of a mixing fermenter, and separating the mixture fermentation product by means of a iii fermentation product separation device to obtain a iii fermentation product; wherein the third fermentation product is grease.
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