CN114657807B - Method for promoting lignin accumulation in wood pulping prehydrolysis liquid by using biological enzyme - Google Patents
Method for promoting lignin accumulation in wood pulping prehydrolysis liquid by using biological enzyme Download PDFInfo
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- D21C—PRODUCTION OF CELLULOSE BY REMOVING NON-CELLULOSE SUBSTANCES FROM CELLULOSE-CONTAINING MATERIALS; REGENERATION OF PULPING LIQUORS; APPARATUS THEREFOR
- D21C5/00—Other processes for obtaining cellulose, e.g. cooking cotton linters ; Processes characterised by the choice of cellulose-containing starting materials
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Abstract
The invention relates to the technical field of biocatalysis, and discloses a method for promoting lignin accumulation in wood pulping prehydrolysis liquid by using biological enzyme, which comprises the following specific steps: treating the wood pulping prehydrolysis liquid by manganese peroxidase; and (3) regulating the hydrolysate treated by the manganese peroxidase to be acidic, separating precipitate by using a centrifugal or precipitation or filtration mode, and washing the precipitate for a plurality of times by using acidified deionized water to obtain the purified lignin. According to the invention, the lignin in the prehydrolysis liquid is biologically modified by utilizing the catalysis of manganese peroxidase, so that the molecular weight of the lignin is increased to polymerize and deposit the lignin, the subsequent extraction of the lignin is facilitated, and the method is simple and is beneficial to promoting the separation and high-value utilization of the lignin.
Description
Technical Field
The invention belongs to the technical field of biocatalysis, and particularly relates to a method for promoting lignin accumulation in wood pulping prehydrolysis liquid by using biological enzymes.
Background
In the production process of dissolving pulp, most hemicellulose and part lignin can be removed by pretreating pulping raw materials by a hydrothermal method, so that the lignin removal efficiency of subsequent pulping is improved. Under the conditions of high temperature and high pressure, the hydronium ions generated by water electrolysis provide acidic reaction conditions, so that the linkage bond between hemicellulose and lignin is broken, the hemicellulose is degraded and dissolved out, and the acidic environment can also lead to the breaking of the acetyl side chain of the hemicellulose to form acetic acid, thereby further promoting the self-hydrolysis reaction of the raw materials. The main components in the hydrolysate are hemicellulose, lignin, acetic acid, furfural and other degradation byproducts. Lignin in the hydrolysate can be used for generating fuel and chemicals such as pyrolysis oil, synthesis gas and the like through pyrolysis, gasification, hydrogenation reduction or oxidation and the like, and can also be modified to be used for preparing adhesives, so that the biomass hydrolysate is an important biomass resource.
However, the components of the hydrolysate are complex, and are difficult to directly and efficiently utilize, so that lignin needs to be separated. Lignin separation can be carried out according to the properties of molecular weight, functional groups and the like, lignin with the same structure is separated, lignin with relatively uniform properties is obtained, and the lignin separation is beneficial to realizing the efficient utilization of lignin with different fractions in the downstream process. At present, the separation method mainly comprises an organic solvent method, a membrane method, a gradient precipitation method and the like. The lignin obtained by the organic solvent method has the advantages of high purity, low ash content and the like, but the relatively complex process increases the cost, the organic solvent possibly generates harm to the environment, and the organic solvent method also has the problems of large reagent dosage, difficult recovery, low economy and the like. Although ultrafiltration is widely applied to industrial lignin treatment, lignin can be effectively concentrated and purified, the energy consumption is low, the membrane separation cost is high, the service period of the membrane is short, and the membrane is easy to pollute, block and damage. The precipitation separation mainly comprises two separation modes of acid precipitation and anti-solvent precipitation, and the acid precipitation separation has more advantages in cost, and is a method for extracting lignin commonly used in industry. Therefore, the method adopted at present is difficult to realize high-efficiency separation of lignin, hemicellulose and other components in the hydrolysate, and other medicines are easy to be introduced to improve the cost of biomass refining or cause secondary pollution.
Disclosure of Invention
Aiming at the problems of low lignin separation efficiency, high cost and the like in the existing pulping prehydrolysis liquid, the invention aims to provide a method for promoting lignin accumulation in the wood pulping prehydrolysis liquid by using biological enzyme. The invention polymerizes lignin through biological enzyme modification, the molecular weight becomes larger and deposits, and then the lignin is separated through centrifugation, acid precipitation, filtration and other modes, thereby improving the separation efficiency of lignin, reducing the dosage of chemicals and having great significance for the grading utilization of lignin in hydrolysate.
In order to solve the technical problems, the technical scheme of the invention is as follows:
a method for promoting lignin accumulation in wood pulping prehydrolysis liquid by using biological enzyme comprises the following specific steps:
treating the wood pulping prehydrolysis liquid by manganese peroxidase;
adjusting the pH value of the hydrolysate after manganese peroxidase treatment to be acidic, stirring fully, and separating to obtain a precipitate which is lignin.
Preferably, the wood pulping prehydrolysis liquid is treated by manganese peroxidase, and the specific process is as follows: adding a certain amount of manganese peroxidase into the prehydrolysis liquid, and then sequentially adding a certain amount of MnSO 4 The solution and hydrogen peroxide start reaction, and the reaction is carried out under acidic condition and certain reaction temperature and reaction time.
Further preferably, the wood pulping prehydrolysis liquid is treated by manganese peroxidase, and the manganese peroxidase dosage is 50-800 U.L -1 ，MnSO 4 The dosage of the solution is 0.01-1 mM.L -1 The hydrogen peroxide is used in an amount of 1 to 50mM L -1 The method comprises the steps of carrying out a first treatment on the surface of the The reaction temperature is 10-60 ℃, the reaction time is 0.5-10 h, and the pH value is 2-7.
Preferably, the pH value of the hydrolysate after manganese peroxidase treatment is adjusted to be 2-7.
Preferably, the precipitate is separated by centrifugation or precipitation or filtration, and washed with acidified deionized water.
Further preferably, the pH value of the acidified deionized water is the same as the pH value of the hydrolysate after manganese peroxidase treatment.
Preferably, the raw materials of the wood pulping prehydrolysis liquid are pulp fiber raw materials of fast-growing poplar, acacia and fast-growing eucalyptus broad-leaved wood.
Preferably, the wood pulping prehydrolysis liquid is obtained by pretreating wood pulping fiber raw materials by a hydrothermal method.
The lignin is obtained by the method for promoting the accumulation of lignin in the wood pulping prehydrolysis liquid by using biological enzyme.
The lignin is applied to the fields of preparation of pyrolysis oil, adhesives, adsorbents and the like.
The method utilizes manganese peroxidase to treat lignin in the hydrolysate, so that the lignin is polymerized and modified to be easily separated, and lignin is obtained by centrifugation, precipitation, filtration or the like.
The invention utilizes manganese peroxidase to treat the hydrolysate, changes the property of lignin in the hydrolysate, polymerizes lignin micromolecules, improves the molecular weight of the lignin, promotes the autodeposition process of the lignin, ensures that the lignin is easy to separate, and is a feasible means for promoting the separation and high-value utilization of lignin components.
Compared with the prior art, the invention has the following steps:
the invention provides a method for polymerizing lignin in hydrolysate by taking wood pulping prehydrolysate as a raw material, which utilizes manganese peroxidase to change the property of lignin and improves the molecular weight of lignin so that the lignin is easy to separate. Compared with the traditional lignin separation means, the method is simple, adopts a green and environment-friendly biological enzyme modification technology, improves the molecular weight of lignin, ensures that the lignin is easy to separate, has little chemical consumption, does not change the properties of lignin, hemicellulose and other components in hydrolysate, and has good practical application value and prospect.
Detailed Description
The following description of the present invention will be made clearly and fully, and it is apparent that the embodiments described are some, but not all, of the embodiments of the present invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments in accordance with the present application. As used herein, the singular is also intended to include the plural unless the context clearly indicates otherwise, and furthermore, it is to be understood that the terms "comprises" and/or "comprising" when used in this specification are taken to specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof.
As mentioned before, the main components of the prehydrolysis liquid obtained in the prehydrolysis process of the wood pulping fiber raw material are hemicellulose, lignin and other degradation products. On one hand, lignin is used as an important raw material for biomass refining, and has high utilization value; on the other hand, the existence of lignin has serious impediments to the high-value utilization of saccharides in the hydrolysate, so that the waste of resources and the loss of economic benefits of enterprises are caused, and the lignin has great significance for the efficient separation of lignin in the hydrolysate. The invention utilizes the biological enzyme method to modify lignin in the hydrolysate, improves the molecular weight of lignin, further promotes the deposition of lignin, and provides reference and technical support for the efficient separation and utilization of lignin in the hydrolysate.
The embodiment provides a method for promoting lignin accumulation in wood pulping prehydrolysis liquid by using biological enzyme, wherein the raw materials of the wood pulping prehydrolysis liquid are fast-growing poplar, acacia, fast-growing eucalyptus broadleaf wood pulping fiber raw materials, and the wood pulping prehydrolysis liquid is obtained by pretreating the wood pulping fiber raw materials by a hydrothermal method. The method comprises the following specific steps:
(1) The method for treating the wood pulping prehydrolysis liquid by using manganese peroxidase comprises the following specific processes: adding a certain amount of manganese peroxidase into the prehydrolysis liquid, and then sequentially adding a certain amount of MnSO 4 The solution and hydrogen peroxide start reaction, and the reaction is carried out under acidic condition, certain reaction temperature and reaction time; wherein the manganese peroxidase dosage is 50-800 U.L -1 Preferably 375 U.L -1 ；MnSO 4 The dosage of the solution is 0.01-1 mM.L -1 Preferably 0.5 mM.L -1 The method comprises the steps of carrying out a first treatment on the surface of the Passing throughThe amount of hydrogen oxide is 1-50 mM.L -1 Preferably 4.4 mM.L -1 、6.88mM·L -1 The method comprises the steps of carrying out a first treatment on the surface of the The reaction temperature is 10-60 ℃, preferably 30 ℃; the reaction time is 0.5-10 h, preferably 4h and 5h; the pH is 2 to 7, preferably 3.6 to 6.0, more preferably 4.5.
(2) The pH value of the hydrolysate after the manganese peroxidase treatment is regulated to 2-7, preferably the pH value is 2, and the hydrolysate is fully stirred, and the precipitate obtained by separation is lignin. Specifically, the precipitate is separated by centrifugation or precipitation or filtration, and is washed by acidified deionized water with the same pH value as the pH value of the hydrolysate after manganese peroxidase treatment.
For a better understanding of the above-described embodiments of the present invention, they are further described below with reference to specific examples.
In the following examples and comparative examples:
the raw materials of the wood pulping prehydrolysis liquid are pulp fiber raw materials of fast-growing poplar, acacia, fast-growing eucalyptus broad-leaved wood.
The wood pulping prehydrolysis liquid is obtained by pretreating wood pulping fiber raw materials by a hydrothermal method.
The manganese peroxidase is commercial brown solid powder manganese peroxidase.
Example 1:
the method for promoting the accumulation of lignin in wood pulping prehydrolysis liquid by using biological enzyme comprises the following steps:
(1) Treating the prehydrolysis liquid of the fast-growing poplar pulp by manganese peroxidase, and adding 375 U.L into the prehydrolysis liquid -1 Manganese peroxidase, then added sequentially with 0.5 mM.L -1 MnSO 4 Solution and 6.88 mM.L -1 The hydrogen peroxide was allowed to react, the reaction temperature was controlled at 30℃and pH 4.5, the shaking table rotation speed was 150rpm, and after 6 hours of reaction, the enzyme was inactivated by a boiling water bath for 5 minutes to terminate the reaction.
(2) Adjusting the pH value of the hydrolysate treated by manganese peroxidase to 2 by using an HCl solution, magnetically stirring for 30min, centrifuging for 15min at a rotating speed of 10000rpm, and washing for 2-3 times by using deionized water with the pH value of 2 to obtain lignin.
Example 2:
the method for promoting the accumulation of lignin in wood pulping prehydrolysis liquid by using biological enzyme comprises the following steps:
(1) Treating the pretrolysis liquid of the acacia wood pulp by manganese peroxidase, and adding 375 U.L into the pretrolysis liquid -1 Manganese peroxidase, then added sequentially with 0.5 mM.L -1 MnSO 4 Solution and 4.4 mM.L -1 The hydrogen peroxide was allowed to react, the reaction temperature was controlled at 30℃and pH 4.5, the shaking table rotation speed was 150rpm, and after 5 hours of reaction, the enzyme was inactivated by a boiling water bath for 5 minutes to terminate the reaction.
(2) Adjusting the pH value of the hydrolysate treated by the manganese peroxidase to 2 by using an HCl solution, magnetically stirring for 30min, centrifuging for 15min at a rotating speed of 10000rpm, and washing for 2-3 times by using deionized water with the pH value of 2 to obtain lignin.
Blank examples:
the preparation process was the same as in example 1, except that the addition amount of manganese peroxidase was 0.
Comparative example 1:
the pH value of the initial fast-growing poplar pulp pre-hydrolysate which is not subjected to enzyme treatment in the embodiment 1 is adjusted to 2 by using HCl solution, the solution is magnetically stirred for 30min, and then centrifuged for 15min at 10000rpm, and the solution is washed for 2 to 3 times by using deionized water with the pH value of 2, so that lignin is obtained.
Comparative example 2:
the pH value of the initial acacia senegal pulp pre-hydrolysis liquid which is not subjected to enzyme treatment in the embodiment 2 is adjusted to be 2 by utilizing HCl solution, the mixture is stirred for 30min by utilizing magnetic force, and then the mixture is centrifuged for 15min at the rotating speed of 10000rpm, and the mixture is washed for 2 to 3 times by utilizing deionized water with the pH value of 2, so that lignin is obtained.
Comparative example 3:
the method for promoting lignin accumulation in wood pulping prehydrolysis liquid by utilizing horseradish peroxidase comprises the following steps:
(1) Treating the prehydrolysis liquid of the fast-growing poplar pulp by horseradish peroxidase, and adding 900 U.L to the prehydrolysis liquid -1 Horseradish peroxidase, 30 mM.L -1 Hydrogen peroxide initiationThe reaction was stopped by controlling the reaction temperature to 30℃and pH 7, shaking the reaction at 150rpm, and inactivating the enzyme in a boiling water bath for 5min after 5h of reaction.
(2) Regulating the pH value of the hydrolysate treated by horseradish peroxidase to 2 by using an HCl solution, magnetically stirring for 30min, centrifuging for 15min at a rotating speed of 10000rpm, and washing for 2-3 times by using deionized water with the pH value of 2 to obtain lignin.
Comparative example 4:
the method for promoting lignin accumulation in wood pulping prehydrolysis liquid by laccase comprises the following steps:
(1) Treating the prehydrolysis liquid of the fast-growing poplar pulp by laccase, and adding 1000U.L to the prehydrolysis liquid -1 Laccase, controlling the reaction temperature to be 30 ℃, controlling the pH value to be 5.5, rotating the shaking table to be 150rpm, and inactivating enzyme in boiling water bath for 5min after reacting for 3h to terminate the reaction.
(2) Regulating the pH value of the hydrolysis liquid treated by laccase to 2 by using an HCl solution, magnetically stirring for 30min, centrifuging for 15min at a rotating speed of 10000rpm, and washing for 2-3 times by using deionized water with the pH value of 2 to obtain lignin.
The lignin obtained in example 1, example 2, blank examples and comparative examples 1 to 4 was measured for the molecular weight of each lignin, and the measured data are shown in table 1 below.
Molecular weight of lignin obtained in Table 1
From the data in table 1, it is clear from the above examples that the weight average molecular weight (Mw) and the number average molecular weight (Mn) of lignin in the hydrolysate can be increased after the treatment of the hydrolysate with manganese peroxidase. Under the reaction conditions of example 1, after the fast-growing poplar prehydrolysis liquid is subjected to manganese peroxidase treatment, the lignin weight average molecular weight and the number average molecular weight are respectively increased from 1643 and 976 to 3066 and 1410, and the weight average molecular weight and the number average molecular weight after enzyme treatment are respectively 1.87 times and 1.44 times that of untreated; under the reaction conditions of example 2, lignin weight average molecular weight and number average molecular weight increased from 1431 and 900 to 2760 and 1240, respectively, after the pre-hydrolysate of acacia catechu was subjected to manganese peroxidase treatment, and the weight average molecular weight and number average molecular weight after enzyme treatment were 1.93 times and 1.37 times, respectively, as compared with that of the initial hydrolysate. As can be seen from the molecular weight of lignin in comparative examples 3 and 4, the weight average molecular weight of lignin in the pre-hydrolyzed solution of fast-growing poplar treated with horseradish peroxidase or laccase was 1.28 and 1.20 times that of the initial hydrolyzed solution, respectively, and the number average molecular weight was 1.05 and 1.25 times that of the initial hydrolyzed solution, respectively, and the polymerization effect was lower than that of manganese peroxidase.
In conclusion, the manganese peroxidase can polymerize lignin in the hydrolysate, regulate and control the molecular weight of lignin, so that lignin is easy to separate by means of acid precipitation, centrifugation, filtration and the like, and the polymerization effect is superior to that of biological enzymes such as laccase, horseradish peroxidase and the like, thus the lignin separation method is an environment-friendly lignin separation means.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the invention.
Claims (8)
1. A method for promoting lignin accumulation in wood pulping prehydrolysis liquid by using biological enzyme is characterized by comprising the following specific steps:
treating the wood pulping prehydrolysis liquid by manganese peroxidase; the specific process of treating wood pulping prehydrolysis liquid by manganese peroxidase is as follows: adding a certain amount of manganese peroxidase into the prehydrolysis liquid, and then sequentially addingQuantitative MnSO 4 The solution and hydrogen peroxide start reaction, and the reaction is carried out under the acidic condition and at a certain reaction temperature for a certain reaction time;
adjusting the pH value of the hydrolysate after manganese peroxidase treatment to be acidic, stirring fully, and separating to obtain a precipitate which is lignin;
wherein the manganese peroxidase dosage is 50-800 U.L -1 ，MnSO 4 The dosage of the solution is 0.01-1 mM.L -1 The hydrogen peroxide is used in an amount of 1 to 50mM L -1 The method comprises the steps of carrying out a first treatment on the surface of the The reaction temperature is 10-60 ℃, the reaction time is 0.5-10 h, and the pH value is 2-7.
2. The method for promoting lignin accumulation in wood pulping prehydrolysis liquid by utilizing biological enzymes according to claim 1, wherein the pH value of the hydrolysis liquid after manganese peroxidase treatment is adjusted to be 2-7.
3. The method for promoting lignin accumulation in wood pulping prehydrolysis liquid by utilizing biological enzymes according to claim 1, wherein the precipitate is separated by adopting a centrifugal or precipitation or filtration mode, and is washed by adopting acidified deionized water.
4. The method for promoting lignin accumulation in wood pulping pre-hydrolysate using biological enzyme according to claim 3 wherein the pH of the acidified deionized water is the same as the pH of the hydrolysate after manganese peroxidase treatment.
5. The method for promoting lignin accumulation in wood pulping prehydrolysis liquid by utilizing biological enzymes according to claim 1, wherein the raw materials of the wood pulping prehydrolysis liquid are pulp fiber raw materials of fast-growing poplar, acacia and fast-growing eucalyptus broad-leaved wood.
6. The method for promoting lignin accumulation in wood pulping prehydrolysis liquid by utilizing biological enzymes according to claim 1, wherein the wood pulping prehydrolysis liquid is obtained by pretreating wood pulping fiber raw materials by a hydrothermal method.
7. Lignin, characterized in that it is obtained by the method for promoting the accumulation of lignin in wood pulping prehydrolysis liquor using biological enzymes according to any one of claims 1 to 6.
8. Use of lignin according to claim 7 for the preparation of pyrolysis oil, adhesives, adsorbents.
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