CN101298620B - Method for preparing ethanol by one-step fermentation after ion liquid stage treatments from native cellulose material - Google Patents
Method for preparing ethanol by one-step fermentation after ion liquid stage treatments from native cellulose material Download PDFInfo
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- IAZSXUOKBPGUMV-UHFFFAOYSA-N 1-butyl-3-methyl-1,2-dihydroimidazol-1-ium;chloride Chemical compound [Cl-].CCCC[NH+]1CN(C)C=C1 IAZSXUOKBPGUMV-UHFFFAOYSA-N 0.000 claims description 2
- YZPNFYQRPJKWFJ-UHFFFAOYSA-N 2-methyl-1h-imidazol-1-ium;chloride Chemical compound Cl.CC1=NC=CN1 YZPNFYQRPJKWFJ-UHFFFAOYSA-N 0.000 claims description 2
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- NAQCUPPQGWEEHC-UHFFFAOYSA-N methanesulfonic acid;1-methylimidazole Chemical compound CS([O-])(=O)=O.C[NH+]1C=CN=C1 NAQCUPPQGWEEHC-UHFFFAOYSA-N 0.000 claims description 2
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 239000011831 acidic ionic liquid Substances 0.000 description 2
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- XLSZMDLNRCVEIJ-UHFFFAOYSA-N methylimidazole Natural products CC1=CNC=N1 XLSZMDLNRCVEIJ-UHFFFAOYSA-N 0.000 description 2
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- 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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Abstract
本发明涉及利用天然纤维素原料经离子液体分级处理一步发酵制备乙醇的方法。其中将天然纤维素原料依次用离子液体A和B处理分别除去半纤维素、木质素后，再以得到的纤维素为底物一步发酵制备乙醇，所述的离子液体A为酸性或碱性离子液体，所述的离子液体B为卤化咪唑类离子液体。本发明通过利用离子液体对天然纤维素原料进行分级处理，把其中的半纤维素、木质素等对发酵生产乙醇有害的物质去除以作他用，仅利用纤维素作为底物进行乙醇发酵，实现天然纤维素原料的全利用。同时可避免半纤维素、木质素等对纤维素酶的吸附，提高酶活；反应器内不发酵的物质积累少，可实现长时间连续发酵，进料浓度高，酵母、纤维素酶循环利用，回收乙醇浓度高，节约用水。The invention relates to a method for preparing ethanol by one-step fermentation using natural cellulose raw materials through ionic liquid classification treatment. Wherein the natural cellulose raw material is sequentially treated with ionic liquids A and B to remove hemicellulose and lignin respectively, and then the obtained cellulose is used as a substrate for one-step fermentation to prepare ethanol, and the ionic liquid A is an acidic or alkaline ion liquid, the ionic liquid B is a halogenated imidazole ionic liquid. The invention uses ionic liquids to classify natural cellulose raw materials, removes hemicellulose, lignin and other substances harmful to fermentation and production of ethanol for other purposes, and only uses cellulose as a substrate to carry out ethanol fermentation. Full utilization of natural cellulose raw materials. At the same time, it can avoid the adsorption of hemicellulose and lignin to cellulase and improve the enzyme activity; the accumulation of non-fermentable substances in the reactor is small, which can realize long-term continuous fermentation, high feed concentration, and recycling of yeast and cellulase , recover ethanol with high concentration and save water.
Description
(一)技术领域(1) Technical field
本发明属于乙醇发酵工业技术领域，特别涉及利用天然纤维素原料经离子液体分级处理一步发酵制备乙醇的方法。The invention belongs to the technical field of ethanol fermentation industry, and particularly relates to a method for preparing ethanol by one-step fermentation using natural cellulose raw materials through ionic liquid classification treatment.
(二)背景技术(2) Background technology
乙醇是生物质液体能源材料的主要代表，也是石化燃料最可能的替代品。随着社会的进步，环境污染、能源短缺问题越来越困扰人类的生存与发展，乙醇作为清洁可再生能源，具有愈益重要的现实意义。以淀粉类和糖类化合物为原料，采用微生物发酵生产乙醇是一项成熟的技术，但其原料成本高昂且消耗并不富余的粮食，难以满足当今社会大规模的开发和利用。Ethanol is the main representative of biomass liquid energy materials and the most likely substitute for fossil fuels. With the progress of society, environmental pollution and energy shortages have increasingly plagued the survival and development of human beings. As a clean and renewable energy source, ethanol has increasingly important practical significance. The production of ethanol by microbial fermentation using starch and sugar compounds as raw materials is a mature technology, but the cost of raw materials is high and the food consumption is not abundant, so it is difficult to meet the large-scale development and utilization in today's society.
纤维素是地球上贮量最丰富的有机物质，植物每年通过光合作用能产生高达1550亿吨纤维素类物质。它成本低廉，来源广泛，目前每年用于工业过程或燃烧的纤维素约占2％左右，还有很大一部分未被利用，因此以纤维素作为发酵原料制备乙醇成为必然的趋势。以天然纤维素原料发酵生产乙醇主要以酿酒酵母为菌种，通过纤维素水解生成葡萄糖后，发酵制得乙醇。该工艺需要解决两大技术问题：(1)天然纤维素的水解产物对发酵的抑制作用；(2)木质纤维素水解产物五碳糖(木糖)、六碳糖(主要是葡萄糖)等混合糖的高效发酵问题。Cellulose is the most abundant organic substance on the earth, and plants can produce up to 155 billion tons of cellulose substances through photosynthesis every year. It is low in cost and has a wide range of sources. At present, about 2% of the cellulose is used in industrial processes or burned every year, and a large part is not utilized. Therefore, it is an inevitable trend to use cellulose as a fermentation raw material to prepare ethanol. Fermentation of natural cellulose raw materials to produce ethanol mainly uses Saccharomyces cerevisiae as the strain, and after cellulose is hydrolyzed to generate glucose, ethanol is produced by fermentation. This process needs to solve two major technical problems: (1) the inhibitory effect of the hydrolyzate of natural cellulose on fermentation; Efficient fermentation of sugars.
纤维素水解的方法主要包括酸解和酶解。前者工艺繁琐，对设备有腐蚀，更重要的是产生大量的对发酵产生抑制的物质(如乳糖醛、酚类化合物等)。酶解较为理想，但纤维素酶价格昂贵，不能循环利用；且由于纤维素是复杂的晶状结构，周围附着半纤维素和木质素，必须经预处理后，纤维素酶才能发生作用。常见的预处理方法有物理法(如机械粉碎、蒸汽爆裂法等)和化学法(如臭氧法、酸法、碱法等)，但都不令人满意。其中蒸汽爆破被认为有应用前景，但它破坏了木聚糖的结构，还可能积累对微生物生长起抑制作用的物质，且需要消耗大量蒸气，另外木质素还不能被除去。出现混合糖发酵的问题是由于半纤维素水解生成了木糖。对于酵母来讲，木糖不能被发酵生成乙醇，且其存在对纤维素酶水解纤维素有抑制作用，随着底物中木糖浓度的增加，乙醇的收率将会下降。The methods of cellulose hydrolysis mainly include acid hydrolysis and enzymatic hydrolysis. The former process is loaded down with trivial details, and equipment is corroded, and what is more important is to produce a large amount of substances (such as lactose aldehyde, phenolic compound etc.) that produce inhibition to fermentation. Enzymatic hydrolysis is ideal, but cellulase is expensive and cannot be recycled; and because cellulose is a complex crystalline structure with hemicellulose and lignin attached around it, cellulase must be pretreated before it can work. Common pretreatment methods include physical methods (such as mechanical crushing, steam explosion methods, etc.) and chemical methods (such as ozone methods, acid methods, alkali methods, etc.), but none of them are satisfactory. Among them, steam explosion is considered to have application prospects, but it destroys the structure of xylan, may also accumulate substances that inhibit microbial growth, and requires a large amount of steam consumption, and lignin cannot be removed. The problem with mixed sugar fermentation occurs due to the hydrolysis of hemicellulose to xylose. For yeast, xylose cannot be fermented to produce ethanol, and its presence can inhibit the hydrolysis of cellulose by cellulase. As the concentration of xylose in the substrate increases, the yield of ethanol will decrease.
木质纤维素发酵生产乙醇的方法中目前比较流行的是同步糖化发酵，该方法优点在于解除了葡萄糖对纤维素酶的抑制作用，提高了酶解效率，减少了纤维素酶的用量，所需要的反应设备减少，污染的可能性降低。但随着发酵的进行，乙醇的大量积累会对微生物的活性产生很大的影响，同时也对纤维素酶产生抑制作用。Synchronous saccharification and fermentation is currently the most popular method of lignocellulose fermentation to produce ethanol. The advantage of this method is that it relieves the inhibitory effect of glucose on cellulase, improves the efficiency of enzymatic hydrolysis, reduces the amount of cellulase, and requires Reaction equipment is reduced and the possibility of contamination is reduced. However, as the fermentation proceeds, the large accumulation of ethanol will have a great impact on the activity of microorganisms and also inhibit cellulase.
(三)发明内容(3) Contents of the invention
本发明的目的在于提供一种天然纤维素原料经离子液体分级处理一步发酵制备乙醇的方法，解决混合糖发酵以及其引起的一系列问题，最终解决乙醇生产效率低的问题。The purpose of the present invention is to provide a method for preparing ethanol by one-step fermentation of natural cellulose raw materials through ionic liquid classification, to solve the mixed sugar fermentation and a series of problems caused by it, and finally solve the problem of low ethanol production efficiency.
本发明采用的技术方案如下：The technical scheme that the present invention adopts is as follows:
天然纤维素原料经离子液体分级处理一步发酵制备乙醇的方法，其中将天然纤维素原料先依次用离子液体A和B处理分别除去半纤维素、木质素，再以得到的纤维素为底物发酵制备乙醇，所述的离子液体A为酸性或碱性离子液体，其中酸性离子液体优选带磺酸根基团的离子液体；所述的离子液体B为卤化咪唑类离子液体。A method for preparing ethanol by one-step fermentation of natural cellulose raw materials through ionic liquid classification treatment, wherein the natural cellulose raw materials are firstly treated with ionic liquids A and B to remove hemicellulose and lignin respectively, and then fermented with the obtained cellulose as a substrate To prepare ethanol, the ionic liquid A is an acidic or basic ionic liquid, wherein the acidic ionic liquid is preferably an ionic liquid with a sulfonate group; the ionic liquid B is a halogenated imidazole ionic liquid.
本发明的关键在于先将天然纤维素原料中含有的对发酵过程有影响的半纤维素和木质素分别去除，采用的手段是分别使用离子液体A和离子液体B，酸性或碱性的离子液体A要求对天然纤维素原料中的半纤维素有很好的溶解性，但是不溶解纤维素和木质素；卤化咪唑类离子液体B对纤维素和木质素的溶解度要有较大差别。相比较于物理或者化学的预处理方法，本处理方法简单，且分离效果好；相较于无机或有机类的溶剂，使用离子液体不仅环保，且经简单处理后即可循环使用，降低成本。The key of the present invention is to firstly remove the hemicellulose and lignin contained in the natural cellulose raw material that have an impact on the fermentation process, and the means used are to use ionic liquid A and ionic liquid B, acidic or alkaline ionic liquid respectively A requires good solubility for hemicellulose in natural cellulose raw materials, but does not dissolve cellulose and lignin; halogenated imidazole ionic liquid B has a large difference in solubility for cellulose and lignin. Compared with physical or chemical pretreatment methods, this treatment method is simple and has good separation effect; compared with inorganic or organic solvents, the use of ionic liquids is not only environmentally friendly, but also can be recycled after simple treatment, reducing costs.
较好的，天然纤维素原料用离子液体处理的过程可如下进行：将天然纤维素原料粉碎至20～200目，加入天然纤维素原料质量2-20倍的离子液体A在60-160℃下浸泡2-10小时，分离出固形物，得到除去半纤维素的天然纤维素；将除去半纤维素的天然纤维素再与离子液体B按照质量比1∶2-20混合，于60-130℃加热0.5-4h，将固形物分离除去，得除去木质素的纤维素溶液，处理后得纤维素。Preferably, the process of treating natural cellulose raw materials with ionic liquids can be carried out as follows: crush the natural cellulose raw materials to 20-200 meshes, add ionic liquid A 2-20 times the mass of the natural cellulose raw materials at 60-160 ° C Soak for 2-10 hours, separate the solid matter, and obtain natural cellulose without hemicellulose; mix the natural cellulose with ionic liquid B at a mass ratio of 1:2-20, and store at 60-130°C Heating for 0.5-4h, separating and removing the solids to obtain a lignin-removed cellulose solution, and obtain cellulose after treatment.
以上过程中用离子液体A处理后分离出固形物后的溶液即为半纤维素的离子液体溶液，加入水或乙醇后半纤维素即可沉淀出来，离心后半纤维素可作他用，离子液体A可通过旋转蒸发后再生并循环使用。卤化咪唑类离子液体B是可以溶解半纤维素和纤维素的，而对木质素溶解度极小，在半纤维素已经除去的情况下，离子液体B中溶解的只有纤维素，而木质素作为固形物在分离时被除去。具体的，溶解有纤维素的离子液体可以通过加入乙醇或水使纤维素沉淀出来，而离子液体通过旋转蒸发后再生并循环使用。In the above process, the solution obtained after the solid matter is separated after treatment with ionic liquid A is the ionic liquid solution of hemicellulose. After adding water or ethanol, the hemicellulose can be precipitated, and the hemicellulose can be used for other purposes after centrifugation. Liquid A can be regenerated and recycled by rotary evaporation. Halogenated imidazole ionic liquid B can dissolve hemicellulose and cellulose, but has very little solubility for lignin. When hemicellulose has been removed, only cellulose is dissolved in ionic liquid B, and lignin is a solid are removed during separation. Specifically, the ionic liquid in which cellulose is dissolved can be precipitated by adding ethanol or water, and the ionic liquid can be regenerated and recycled by rotary evaporation.
天然纤维素原料经离子液体分级处理后，木质素与半纤维素已被分离除去，这就避免了后续糖化发酵过程中水解产物的混合糖发酵问题，以及分解不完全对纤维素酶的抑制作用，提高了酶活以及发酵效率，另外发酵过程中累积的不发酵物质减少，可长时间连续发酵。After the natural cellulose raw material is graded by ionic liquid, lignin and hemicellulose have been separated and removed, which avoids the problem of mixed sugar fermentation of hydrolyzate in the subsequent saccharification and fermentation process, and the inhibition of cellulase by incomplete decomposition , improve the enzyme activity and fermentation efficiency, in addition, the accumulation of non-fermentable substances in the fermentation process is reduced, and it can be fermented continuously for a long time.
纤维素酶发酵生产乙醇已经是较为成熟的技术，本发明根据具体情况又进一步提供了较为优选的方法：将纤维素与水按照1-4∶10的质量比例混合，灭菌后加入纤维素酶，于40～60℃下保温20～40小时进行酶解，然后再于30～40℃，无菌接入酿酒酵母发酵10～30小时，生成乙醇。The production of ethanol by cellulase fermentation is a relatively mature technology, and the present invention further provides a more preferred method according to specific conditions: mix cellulose and water according to the mass ratio of 1-4:10, add cellulase after sterilization , incubating at 40-60°C for 20-40 hours for enzymolysis, and then aseptically inserting Saccharomyces cerevisiae at 30-40°C for 10-30 hours of fermentation to generate ethanol.
本方法中所述酶解与发酵过程优选在同一反应器中进行，酶解与发酵过程中反应器底部通入CO2。特别是当反应器中乙醇的体积浓度达到3～5％时，增大CO2通气量至乙醇可从反应器中移出。此操作可以解决乙醇的大量积累对微生物的活性产生的影响，同时也避免了乙醇积累对纤维素酶产生的抑制作用。The enzymolysis and fermentation processes in this method are preferably carried out in the same reactor, and CO 2 is introduced into the bottom of the reactor during the enzymolysis and fermentation processes. Especially when the volume concentration of ethanol in the reactor reaches 3-5%, increase the CO 2 gas flow until the ethanol can be removed from the reactor. This operation can solve the impact of a large amount of ethanol accumulation on the activity of microorganisms, and also avoid the inhibitory effect of ethanol accumulation on cellulase production.
在上述纤维素发酵制备乙醇的过程中，反应物料的加入、酶解、发酵、乙醇的分离可连续进行。而由CO2移出的乙醇通过冷凝、吸附、吸收或膜分离回收，可得到体积百分比浓度为30～60％的乙醇。In the process of preparing ethanol by fermenting cellulose, the addition of reaction materials, enzymatic hydrolysis, fermentation and separation of ethanol can be carried out continuously. The ethanol removed from CO2 is recovered by condensation, adsorption, absorption or membrane separation, and ethanol with a concentration of 30-60% by volume can be obtained.
本发明提到的天然纤维素原料优选绿色植物通过光合作用形成的可再生资源，如玉米秸秆、小麦秸秆或稻草等农业废弃物，既保护环境又能废物利用。The natural cellulose raw material mentioned in the present invention is preferably a renewable resource formed by green plants through photosynthesis, such as agricultural waste such as corn stalks, wheat stalks or rice straw, which not only protects the environment but also can be used for waste utilization.
本发明相对于现有技术，有以下优点：Compared with the prior art, the present invention has the following advantages:
本发明通过利用环境友好的离子液体对天然纤维素原料进行分级处理，把其中的半纤维素、木质素等对发酵生产乙醇有害的物质去除以作他用，仅利用纤维素作为底物进行乙醇发酵，实现天然纤维素原料的全利用。同时可避免半纤维素、木质素等对纤维素酶的吸附，提高酶活；反应器内不发酵的物质积累少，可实现长时间连续发酵，进料浓度高，酵母、纤维素酶循环利用，回收乙醇浓度高，节约用水。In the present invention, the natural cellulose raw material is graded by using environment-friendly ionic liquid, and the hemicellulose, lignin and other substances harmful to the fermentation and production of ethanol are removed for other purposes, and only cellulose is used as the substrate for ethanol production. Fermentation to realize the full utilization of natural cellulose raw materials. At the same time, it can avoid the adsorption of hemicellulose and lignin to cellulase and improve the enzyme activity; the accumulation of non-fermentable substances in the reactor is small, which can realize long-term continuous fermentation, high feed concentration, and recycling of yeast and cellulase , recover ethanol with high concentration and save water.
(四)具体实施方式：(4) Specific implementation methods:
以下以具体实施例来说明本发明的技术方案，但本发明的保护范围不限于此：The technical scheme of the present invention is described below with specific examples, but protection scope of the present invention is not limited thereto:
实施例1Example 1
将一定量的玉米秸秆粉碎至40目，按1∶6的质量比例加入离子液体氯化1-甲基-3-(苯基磺酸基)甲基咪唑(酸性)中，在80℃下浸泡4小时。原料部分溶解，经离心，获得主要成分为纤维素和木质素的固形物。溶解在离子液体氯化1-甲基-3-(苯基磺酸基)甲基咪唑中的半纤维，可通过加入水沉淀出来，离心分离；离子液体氯化1-甲基-3-(苯基磺酸基)甲基咪唑可通过真空干燥循环使用。Crush a certain amount of corn stalks to 40 mesh, add the ionic liquid 1-methyl-3-(phenylsulfonic acid group) methylimidazole chloride (acidic) according to the mass ratio of 1:6, soak at 80 ° C 4 hours. The raw material is partially dissolved and centrifuged to obtain a solid whose main components are cellulose and lignin. The hemifiber dissolved in the ionic liquid chloride 1-methyl-3-(phenylsulfonic acid group) methylimidazole can be precipitated by adding water and centrifuged; the ionic liquid chloride 1-methyl-3-( Phenylsulfonate)methylimidazole can be recycled by vacuum drying.
含有纤维素和木质素的固形物，按6∶100的质量比例加入离子液体氯化1-烯丙基-3-甲基咪唑中，在110℃下油浴加热2小时。固形物部分溶解，经离心，获得主要成分为木质素的固形物。溶解于离子液体氯化1-烯丙基-3-甲基咪唑的纤维素，通过加入乙醇溶剂沉淀出来，离心分离；离子液体氯化1-烯丙基-3-甲基咪唑可通过旋转蒸发再生循环使用。The solid matter containing cellulose and lignin was added to the ionic liquid 1-allyl-3-methylimidazole chloride at a mass ratio of 6:100, and heated in an oil bath at 110° C. for 2 hours. The solid is partially dissolved, and centrifuged to obtain a solid whose main component is lignin. The cellulose dissolved in the ionic liquid 1-allyl-3-methylimidazole chloride is precipitated by adding ethanol solvent and centrifuged; the ionic liquid 1-allyl-3-methylimidazole chloride can be obtained by rotary evaporation Recycled use.
分离所得纤维素与水混合配制质量百分比为20％的混合液，加入气升式反应器中，经灭菌后加入纤维素酶酶解，酶加入量为100U/g纤维素，于55℃下保温24小时，生成纤维素酶解物，在该过程中，从反应器底部微通CO2进行搅拌；纤维素酶解物在34℃下，无菌接入酿酒酵母发酵24小时，生成乙醇，该过程中，继续从反应器底部微通CO2进行搅拌；当反应器中乙醇的体积浓度达到3～5％时，增大CO2通气量，进行气提操作，利用CO2的夹带作用，将乙醇从反应器中移出。由CO2移出的乙醇通过冷凝方法(冷媒温度-30℃)回收，最终乙醇体积浓度为40％。The separated cellulose is mixed with water to prepare a mixed solution with a mass percentage of 20%, which is added to an air-lift reactor. After sterilization, cellulase is added for enzymolysis. Incubate for 24 hours to generate cellulose hydrolyzate. During this process, CO 2 is stirred from the bottom of the reactor; the cellulose hydrolyzate is aseptically inserted into Saccharomyces cerevisiae at 34°C for 24 hours to generate ethanol. During this process, continue stirring with CO2 from the bottom of the reactor; when the volume concentration of ethanol in the reactor reaches 3-5%, increase the amount of CO2 aeration to carry out the gas stripping operation, and use the entrainment effect of CO2 , Ethanol was removed from the reactor. The ethanol removed from CO2 is recovered by condensation method (refrigerant temperature -30°C), and the final volume concentration of ethanol is 40%.
实施例2Example 2
将一定量的稻草粉碎至80目，按1∶15的质量比例加入离子液体甲磺酸N-甲基咪唑(酸性)中，在60℃下浸泡6小时。原料部分溶解，经离心，获得主要成分为纤维素和木质素的固形物。半纤维素的分离与离子液体的回收方法同实施例1。A certain amount of rice straw was crushed to 80 meshes, added into the ionic liquid N-methylimidazole methanesulfonate (acidic) at a mass ratio of 1:15, and soaked at 60° C. for 6 hours. The raw material is partially dissolved and centrifuged to obtain a solid whose main components are cellulose and lignin. The separation of hemicellulose and the recovery method of ionic liquid are the same as in Example 1.
含有纤维素和木质素的固形物，按1∶5的质量比例加入离子液体氯化1-丁基-3-甲基咪唑中，微波加热1小时。固形物部分溶解，经离心，获得主要成分为木质素的固形物。纤维素的分离与离子液体的再生方法同实施例1。The solid matter containing cellulose and lignin is added to the ionic liquid 1-butyl-3-methylimidazole chloride at a mass ratio of 1:5, and heated by microwave for 1 hour. The solid is partially dissolved, and centrifuged to obtain a solid whose main component is lignin. The separation of cellulose and the regeneration method of ionic liquid are the same as in Example 1.
分离所得纤维素与水混合配制质量百分比为40％的混合液，加入气升式反应器中，经灭菌后加入纤维素酶酶解，酶加入量为100U/g纤维素，于60℃下保温24小时，生成纤维素酶解物，在该过程中，从反应器底部微通CO2进行搅拌；纤维素酶解物在34℃下，无菌接入酿酒酵母发酵24小时，生成乙醇，该过程中，从反应器底部微通CO2进行搅拌；后续操作同实施例1。由CO2移出的乙醇通过填充活性炭吸附塔回收，待吸附饱和后，加热吸附塔再生，经冷凝最终乙醇浓度为60％。The separated cellulose is mixed with water to prepare a mixed solution with a mass percentage of 40%, which is added to an air-lift reactor, and after sterilization, cellulase is added for enzymolysis. Incubate for 24 hours to generate cellulose hydrolyzate. During this process, CO 2 is stirred from the bottom of the reactor; the cellulose hydrolyzate is aseptically inserted into Saccharomyces cerevisiae at 34°C for 24 hours to generate ethanol. During this process, micro- pass CO from the bottom of the reactor for stirring; subsequent operations are the same as in Example 1. The ethanol removed from CO2 is recovered by filling the activated carbon adsorption tower. After the adsorption is saturated, the adsorption tower is heated for regeneration, and the final ethanol concentration is 60% after condensation.
实施例3Example 3
将一定量的小麦秸秆粉碎至100目，按1∶10的质量比例加入离子液体氢氧化1-甲基-3-正丁基咪唑(碱性)中，在80℃下浸泡8小时。原料部分溶解，经离心，获得主要成分为纤维素和木质素的固形物。半纤维素的分离与离子液体的回收方法同实施例1。A certain amount of wheat straw was crushed to 100 mesh, added to ionic liquid hydroxide 1-methyl-3-n-butylimidazole (alkaline) at a mass ratio of 1:10, and soaked at 80° C. for 8 hours. The raw material is partially dissolved and centrifuged to obtain a solid whose main components are cellulose and lignin. The separation of hemicellulose and the recovery method of ionic liquid are the same as in Example 1.
含有纤维素和木质素的固形物，按6∶100的质量比例加入离子液体溴化1-丁基-3-甲基咪唑中，微波加热3小时。固形物部分溶解，经离心，获得主要成分为木质素的固形物。纤维素的分离与离子液体的再生方法同实施例1。The solid matter containing cellulose and lignin was added to the ionic liquid 1-butyl-3-methylimidazole bromide at a mass ratio of 6:100, and heated by microwave for 3 hours. The solid is partially dissolved, and centrifuged to obtain a solid whose main component is lignin. The separation of cellulose and the regeneration method of ionic liquid are the same as in Example 1.
分离所得纤维素与水混合配制质量百分比为20％的混合液，加入气升式反应器中，经灭菌后加入纤维素酶酶解，酶加入量为100U/g纤维素，于60℃下保温24小时，生成纤维素酶解物，在该过程中，从反应器底部微通CO2进行搅拌；纤维素酶解物在34℃下，无菌接入酿酒酵母发酵24小时，生成乙醇，该过程中，继续从反应器底部微通CO2进行搅拌；当反应器中乙醇的体积浓度达到3～5％时，增大CO2通气量，进行气提操作，将乙醇从反应器中移出。同时向反应器中补加经过灭菌的纤维素与水的混合溶液(质量百分比为40％)，进行连续操作。其他操作同实施例2，最终乙醇浓度为60％。The separated cellulose is mixed with water to prepare a mixed solution with a mass percentage of 20%, which is added to an air-lift reactor. After sterilization, cellulase is added for enzymolysis. Incubate for 24 hours to generate cellulose hydrolyzate. During this process, CO 2 is stirred from the bottom of the reactor; the cellulose hydrolyzate is aseptically inserted into Saccharomyces cerevisiae at 34°C for 24 hours to generate ethanol. During this process, continue stirring with CO2 from the bottom of the reactor; when the volume concentration of ethanol in the reactor reaches 3-5%, increase the amount of CO2 ventilation, carry out air stripping operation, and remove the ethanol from the reactor . At the same time, a mixed solution of sterilized cellulose and water (40% by mass) was added to the reactor for continuous operation. Other operations are the same as in Example 2, and the final ethanol concentration is 60%.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103333924A (en) * | 2013-06-27 | 2013-10-02 | 郑通彪 | Method for producing alcohol by using straw |
Families Citing this family (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101712698B (en) * | 2009-11-25 | 2013-05-01 | 大连工业大学 | Ionic liquid based method for extracting lignin by two-liquid-phase system |
| CN101736060B (en) * | 2010-01-08 | 2012-07-04 | 河南科技大学 | Method for producing ethanol by using ionic liquid |
| CN101824339B (en) * | 2010-05-12 | 2014-05-07 | 哈尔滨理工大学 | Ethanol/water mixed solvent preprocessing biomass and method for preparing liquid fuel |
| CN101871020B (en) * | 2010-07-14 | 2012-05-30 | 河南科技大学 | Method for preparing wood sugar, glucose and lignin with ionic liquid controllable hydrolytic wood fiber raw material |
| CN101985642B (en) * | 2010-12-09 | 2012-09-05 | 东华大学 | Method for preparing bacterial cellulose by using straw |
| CN101985641B (en) * | 2010-12-09 | 2012-09-05 | 东华大学 | Method for preparing bacterial cellulose by using wheat straw |
| CN102154412A (en) * | 2011-01-17 | 2011-08-17 | 同济大学 | Method for increasing sugar yield of cellulase enzymolysis through pretreatment by using ion liquid |
| JP5589871B2 (en) * | 2011-01-27 | 2014-09-17 | トヨタ自動車株式会社 | Method for treating cellulosic biomass, method for producing sugar, alcohol or organic acid from cellulosic biomass |
| CN102321055A (en) * | 2011-09-30 | 2012-01-18 | 中国科学院过程工程研究所 | Method for preparing 5-hydroxymethylfurfural from woody biomasses |
| US11118308B2 (en) | 2013-03-15 | 2021-09-14 | Ip2Ipo Innovations Limited | Treatment of lignocellulosic biomass with ionic liquid |
| BR112015026223B1 (en) | 2013-04-15 | 2022-02-22 | Metsã Spring Oy | Method of producing regenerated cellulose and regenerated hemicellulose, method of treating fiber pulp and method of fractionating hemicellulose and cellulose in fiber pulp |
| CN103469662B (en) * | 2013-09-23 | 2015-08-12 | 福建农林大学 | A kind of method applied ionic liquid-catalyzed hydrolysis and prepare nano-cellulose |
| CN103755971A (en) * | 2013-12-10 | 2014-04-30 | 江南大学 | Method for extracting lignin of straws by acidic ionic liquid and organic solvent |
| CN106750548A (en) * | 2015-11-20 | 2017-05-31 | 中国科学院理化技术研究所 | Activation pretreatment cellulose thermoplastic material and preparation method thereof |
| GB201520453D0 (en) | 2015-11-20 | 2016-01-06 | Imp Innovations Ltd | Process |
| CN106755189B (en) * | 2016-10-31 | 2021-01-05 | 广东工业大学 | Green method for improving enzymolysis efficiency of cellulose in rice straw by utilizing two deep eutectic solvents for step-by-step pretreatment |
| CN112442924B (en) * | 2019-09-03 | 2022-06-14 | 中国科学院大连化学物理研究所 | Method for preparing high-hardness environment-friendly material by using protogenic plant biomass as raw material |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1327067A (en) * | 2000-06-02 | 2001-12-19 | 唐山市冀东溶剂厂 | Method for producing alcohol by using plant fiber as raw material |
| CN1806945A (en) * | 2005-01-20 | 2006-07-26 | 中国科学院过程工程研究所 | Method for complete enzymolysis of straw cellulose by utilizing pretreatment and enzymolysis process |
-
2008
- 2008-05-09 CN CN2008100957987A patent/CN101298620B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1327067A (en) * | 2000-06-02 | 2001-12-19 | 唐山市冀东溶剂厂 | Method for producing alcohol by using plant fiber as raw material |
| CN1806945A (en) * | 2005-01-20 | 2006-07-26 | 中国科学院过程工程研究所 | Method for complete enzymolysis of straw cellulose by utilizing pretreatment and enzymolysis process |
Non-Patent Citations (3)
| Title |
|---|
| Richard P.Swatloski et al.Dissolution of Cellose with Ionic Liquids.《J.AM.CHEM.SOC》.2002,第124卷(第18期),4974-4975. * |
| 刘振 等.稻谷生料发酵生产乙醇研究.《化学工程》.2006,第34卷(第3期),49-52. |
| 刘振等.稻谷生料发酵生产乙醇研究.《化学工程》.2006,第34卷(第3期),49-52. * |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103333924A (en) * | 2013-06-27 | 2013-10-02 | 郑通彪 | Method for producing alcohol by using straw |
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