CN110734943A - Method for improving enzymolysis saccharification effect by pretreating bagasse - Google Patents

Method for improving enzymolysis saccharification effect by pretreating bagasse Download PDF

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CN110734943A
CN110734943A CN201911017844.6A CN201911017844A CN110734943A CN 110734943 A CN110734943 A CN 110734943A CN 201911017844 A CN201911017844 A CN 201911017844A CN 110734943 A CN110734943 A CN 110734943A
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CN110734943B (en
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王朝宇
迟雪文
毕艳红
张睿
杨荣玲
刁怡涵
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Cofco Chongzuo Sugar Co ltd
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Huaiyin Institute of Technology
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Abstract

The invention discloses a method for improving enzymatic saccharification effect by bagasse pretreatment, which comprises the following steps of weighing bagasse, adding a Deep Eutectic Solvent (DES) for pretreatment, carrying out solid-liquid separation after the reaction is finished, adding a KOH solution into the separated solid for pretreatment, carrying out solid-liquid separation after the reaction is finished, respectively collecting solid and liquid parts, washing the solid part to be neutral to obtain the pretreated bagasse, carrying out enzymatic saccharification, and determining the glucose concentration in an enzymatic hydrolysate.

Description

Method for improving enzymolysis saccharification effect by pretreating bagasse
Technical Field
The invention belongs to the technical field of biochemical engineering, and particularly relates to a method for improving enzymolysis saccharification effect by pretreating bagasse.
Background
The biomass energy is used as type typical representative of renewable energy, and has a relatively mature development system in developed countries such as Europe and the United states in the aspects of biological liquid fuel, biomass power generation and the like, so that if the lignocellulosic biomass raw material can be processed and pretreated to produce biomass fuel or a large amount of chemicals with high added value, the global energy crisis can be relieved to a certain extent in , and the pressure of fossil energy is relieved.
Sugarcane is a herbaceous plant with years or more of years, which is common in tropical and subtropical regions, is rich in sugar, vitamins, minerals and the like, China is the third sugarcane planting base in the world after Brazil and India, China is the third sugarcane planting base in China, the main production area of sugarcane is distributed in west, Yunnan, east, Hainan, Fujian, Sichuan and Jiangxi, and the like, the yield of sugarcane is as high as 80 metric tons/hectare, which is much higher than that of crops, so that sugarcane can be regarded as a high-efficiency storage rich in solar energy, which is important sugar crops, the produced cane sugar accounts for 80% of the total sugar content in China, which is the consuming country of sucrose, which is second only in Brazil and India, wherein sucrose is the main source of sugar, accounts for more than 92% of the total sugar content, the sugarcane has the highest weight in the sugar industry of China, bagasse is important sugar manufacturing byproducts, which is the total sugar production of cane sugar, or the sugar juice extracted vigorously by the sugar factory, the infiltration method, the cane has the highest cellulose content of cellulose, the cane juice, the cane pulp, the cellulose content of the cane pulp is about 20-30% of cellulose, the cellulose residue of bagasse is generated by-30 stalks, the bagasse is generated by-30 stalks, the bagasse is generated by the bagasse, the biomass, the biomass is used for the biomass, the biomass is used for biomass, the biomass.
Disclosure of Invention
The invention aims to solve the problems in the prior art, and provides a method for improving enzymolysis saccharification effect by pretreating bagasse.
In order to achieve the purpose, the method for improving the enzymatic saccharification effect by pretreating types of bagasse comprises the following steps:
(1) weighing bagasse, adding DES (data encryption standard) for pretreatment, performing solid-liquid separation on the bagasse after the reaction is finished, and respectively collecting solid and liquid parts;
(2) adding KOH solution into the solid separated in the step (1) for pretreatment, and carrying out solid-liquid separation after the reaction is finished;
(3) and (3) washing the solid part separated in the step (2) to be neutral to obtain pretreated bagasse, carrying out enzymolysis saccharification, and determining the concentration of glucose in the enzymolysis liquid.
The bagasse used in the invention is purchased from Futaste Co. Ltd. (China) or other commercial companies, or the extraction method is according to the documents of Wang Yonggarden, Libuhua, Liuyu ring and the like, the latest progress of the comprehensive utilization technology of the bagasse [ J ] Chinese agricultural bulletin, 2010,26(16):370-375, and the effect is .
Wherein the DES consists of choline chloride: lactic acid was added in a molar ratio of 1: 1-5. Preferably, the DES consists of choline chloride: lactic acid was added in a molar ratio of 1:2, and preparing the product.
the DES is prepared by adding choline chloride and lactic acid into a reaction flask, magnetically stirring at 60-80 deg.C until the mixture becomes colorless clear liquid, transferring to a vacuum drying oven, and adding P2O5Fully drying the mixture for later use.
Wherein, the conditions of DES pretreatment of bagasse are as follows: the pretreatment time is 0.5-6h, the pretreatment solid-to-liquid ratio is 1:8-1:30g/mL, and the pretreatment temperature is 60-130 ℃.
Wherein the mass fraction of the KOH solution is 5 to 40 wt.%.
Wherein, the pretreatment conditions of the KOH solution are as follows: the treatment time is 0.5-5h, the treatment solid-liquid ratio is 1:6-1:15g/mL, and the treatment temperature is 50-100 ℃.
And (3) carrying out solid-liquid separation on the bagasse subjected to KOH pretreatment, respectively collecting solid and liquid parts, washing solid residues to be neutral by using tap water, and then carrying out enzymatic saccharification.
, adding citric acid buffer solution to adjust pH, adding sodium azide solution, adding cellulase, and measuring glucose concentration in the enzymolysis solution by HPLC after reaction.
Preferably, the cellulase is added in an amount of 3-30 FPU/g-substrate (i.e. pretreated bagasse), with a solids loading of 2-20 wt.%, and is subjected to enzymatic hydrolysis at pH 4.8 and a temperature of 50 ℃ for 2-96 h. If the enzymatic hydrolysis reaction volume is 20mL, 1g of pretreated bagasse (dry by absolute) is added at a solids loading of 5 wt.%, and the total reaction volume is 20mL x 5% solids loading (solids content) is 1g of dry by absolute weight.
According to IUPAC standard, the definition of Filter paper enzyme activity (FPA) is 1g of solid enzyme (or 1mL of liquid enzyme), and the enzyme dosage required for generating 1 mu mol of glucose from a Filter paper substrate per hour at 50 ℃ and pH 4.8 is defined as enzyme activity units expressed by U/g (or U/mL).
After the enzymatic saccharification process is finished, quantitative enzymatic hydrolysate is taken to be subjected to High Performance Liquid Chromatography (HPLC) detection, and the content of reducing sugar in the enzymatic hydrolysate is measured, wherein the detection conditions of HPLC are shown in Table 1.
TABLE 1 conditions of liquid chromatography detection
Figure BDA0002246268160000031
The liquid (the black liquid collected in the steps (1) and (2)) generated in the method for improving the enzymolysis and saccharification effect by bagasse pretreatment is applied to the preparation of potassium fertilizer, lignin-based fertilizer and compound fertilizer.
According to the method, the DES and KOH pretreatment is adopted to remove lignin and hemicellulose wrapped on the surface of the cellulose, so that the highly ordered and tightly arranged regions of the cellulose are damaged, pores are generated on the surface of the cellulose, more cellulose is exposed, the porosity and porosity are increased, the accessibility of the cellulose by the cellulase is improved, and the enzymolysis saccharification efficiency is promoted.
Has the advantages that: compared with the prior art, the invention has the following advantages:
the method comprises the steps of pretreating bagasse by DES and KOH, damaging the wrapping of lignin and hemicellulose on cellulose, removing lignin, degrading hemicellulose, changing the crystal structure of cellulose, improving the accessibility and the porosity of cellulose, promoting the contact of cellulase and a substrate, and further improving the yield of cellulase hydrolysis and the yield of reducing sugar, wherein under the pretreatment and enzymatic hydrolysis conditions of the bagasse, the optimal conditions of DES pretreatment are determined as choline chloride and lactic acid (1:2), the reaction temperature is 90 ℃, the reaction time is 1h, the solid-liquid ratio is 1:10, the optimal conditions of KOH pretreatment are that the KOH dosage is 15 wt.%, the reaction temperature is 90 ℃, the reaction time is 2h, and the solid-liquid ratio is 1:10, the research shows that under the conditions of 10% of solid content and 20 FPU/g-substrate, the highest glucose concentration is 47.6 +/-1.1 g/L, the xylose concentration is 21.6 +/-0.5 g/L, 5% of solid content is 20 FPU/g-substrate, the highest glucose yield is 87.4 +/-0.7.1 g/L, the highest xylose concentration is 21.6 +/-0.5 g/L, the highest solid content of bagasse can be found according to the theoretical requirement of waste for developing a renewable resource-rich in bagasse, and the resource can be developed according to the total resource of renewable waste.
The method adopts eutectic solvents DES and KOH solution for pretreatment, has mild reaction conditions, reduces the damage to instruments and equipment, and is beneficial to industrial production and application.
Drawings
FIG. 1 is a Scanning Electron Microscope (SEM) atlas before and after bagasse pretreatment;
FIG. 2 shows Fourier Infrared Spectroscopy (FTIR) spectra before and after bagasse pretreatment;
FIG. 3 is an X-ray diffraction pattern (XRD) before and after bagasse pretreatment;
wherein a is bagasse raw material; b is bagasse after 15 wt.% KOH pretreatment in example 2;
observation of SEM, FTIR and XRD patterns shows that compared with the bagasse raw material which is not pretreated, the bagasse after pretreatment has increased crystallinity, increased porosity and porosity, and increased accessibility of cellulose by cellulase, thereby improving the efficiency of enzymatic saccharification.
Detailed Description
The invention is further illustrated in connection with the following examples and the accompanying drawings.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
In the embodiment, the solid residue part of the sugarcane after extracting the cane sugar is calculated by absolute dry materials; DES is prepared by adding choline chloride and lactic acid into a reaction flask, magnetically stirring at 80 deg.C until the mixture turns into colorless clear liquid, transferring into a vacuum drying oven, and adding P2O5Fully drying the mixture for later use.
Example 1
Weighing 30.0 +/-0.1 g of bagasse, adding 300.0 +/-0.1 g of DES (choline chloride: lactic acid (1: 2)) for pretreatment at 90 ℃ for 1h, carrying out solid-liquid separation after the reaction is finished, then adding 300.0 +/-0.1 mL of KOH solution with the mass fraction of 10%, carrying out pretreatment at 90 ℃ for 2h, and carrying out solid-liquid separation after the reaction is finished; and (4) washing the solid part to be neutral by using tap water, and carrying out an enzymolysis saccharification experiment. The enzymatic hydrolysis reaction volume was 20mL, 1g of pretreated bagasse (absolute dry basis) was added at a 5 wt.% solids loading, the pH of the enzymatic hydrolysis was adjusted to 4.8 by the addition of 0.05mol/L citric acid buffer solution, 0.2mL of sodium azide solution (0.02g/mL) was added, followed by cellulase enzyme at 20 FPU/g-substrate and reaction at 50 ℃ for 72h at 120 rpm. Then, the glucose concentration in the enzymatic hydrolysate was measured by HPLC. The result shows that the concentration of glucose in the enzymolysis liquid is 21.2g/L, the concentration of xylose is 10.2g/L, and the final obtained total sugar yield is 76.3%.
Example 2
30.0 +/-0.1 g of bagasse is weighed, 300.0 +/-0.1 g of DES (choline chloride: lactic acid (1: 2)) is added for pretreatment for 1 hour at 90 ℃, solid-liquid separation is carried out after the reaction is finished, then 300.0 +/-0.1 mL of KOH solution with the mass fraction of 15 percent is added for pretreatment for 2 hours at 90 ℃, and solid-liquid separation is carried out after the reaction is finished. And (4) washing the solid part to be neutral by using tap water, and carrying out an enzymolysis saccharification experiment. The enzymatic hydrolysis reaction volume was 20mL, 1g of pretreated bagasse (absolute dry basis) was added at a 5 wt.% solids loading, the pH of the enzymatic hydrolysis was adjusted to 4.8 by the addition of 0.05mol/L citric acid buffer solution, 0.2mL of sodium azide solution (0.02g/mL) was added, followed by cellulase enzyme at 20 FPU/g-substrate and reaction at 50 ℃ for 72h at 120 rpm. Then, the glucose concentration in the enzymatic hydrolysate was measured by HPLC. The result shows that the concentration of glucose in the enzymolysis liquid is 24.4g/L, the concentration of xylose is 11.0g/L, and the final obtained total sugar yield is 80.6%.
Example 3
30.0 +/-0.1 g of bagasse is weighed, 300.0 +/-0.1 g of DES (choline chloride: lactic acid (1: 2)) is added for pretreatment for 1 hour at 90 ℃, solid-liquid separation is carried out after the reaction is finished, then 300.0 +/-0.1 mL of KOH solution with the mass fraction of 15 percent is added for pretreatment for 2 hours at 70 ℃, and solid-liquid separation is carried out after the reaction is finished. And (4) washing the solid part to be neutral by using tap water, and carrying out an enzymolysis saccharification experiment. The enzymatic hydrolysis reaction volume was 20mL, 1g of pretreated bagasse (absolute dry basis) was added at a 5 wt.% solids loading, the pH of the enzymatic hydrolysis was adjusted to 4.8 by the addition of 0.05mol/L citric acid buffer solution, 0.2mL of sodium azide solution (0.02g/mL) was added, followed by cellulase enzyme at 20 FPU/g-substrate and reaction at 50 ℃ for 72h at 120 rpm. Then, the glucose concentration in the enzymatic hydrolysate was measured by HPLC. The result shows that the concentration of glucose in the enzymolysis liquid is 21.0g/L, the concentration of xylose is 9.5g/L, and the final obtained total sugar yield is 74.7%.
Example 4
30.0 +/-0.1 g of bagasse is weighed, 300.0 +/-0.1 g of DES (choline chloride: lactic acid (1: 2)) is added for pretreatment for 1 hour at 90 ℃, solid-liquid separation is carried out after the reaction is finished, then 300.0 +/-0.1 mL of KOH solution with the mass fraction of 15 percent is added for pretreatment for 2 hours at 100 ℃, and solid-liquid separation is carried out after the reaction is finished. And (4) washing the solid part to be neutral by using tap water, and carrying out an enzymolysis saccharification experiment. The enzymatic hydrolysis reaction volume was 20mL, 1g of pretreated bagasse (absolute dry basis) was added at a 5 wt.% solids loading, the pH of the enzymatic hydrolysis was adjusted to 4.8 by the addition of 0.05mol/L citric acid buffer solution, 0.2mL of sodium azide solution (0.02g/mL) was added, followed by cellulase enzyme at 20 FPU/g-substrate and reaction at 50 ℃ for 72h at 120 rpm. Then, the glucose concentration in the enzymatic hydrolysate was measured by HPLC. The result shows that the concentration of glucose in the enzymolysis liquid is 24.3g/L, the concentration of xylose is 10.4g/L, and the final obtained total sugar yield is 76.0%.
Example 5
30.0 +/-0.1 g of bagasse is weighed, 300.0 +/-0.1 g of DES (choline chloride: lactic acid (1: 2)) is added for pretreatment for 1 hour at 90 ℃, solid-liquid separation is carried out after the reaction is finished, then 300.0 +/-0.1 mL of KOH solution with the mass fraction of 15 percent is added for pretreatment for 1 hour at 90 ℃, and solid-liquid separation is carried out after the reaction is finished. And (4) washing the solid part to be neutral by using tap water, and carrying out an enzymolysis saccharification experiment. The enzymatic hydrolysis reaction volume was 20mL, 1g of pretreated bagasse (absolute dry basis) was added at a 5 wt.% solids loading, the pH of the enzymatic hydrolysis was adjusted to 4.8 by the addition of 0.05mol/L citric acid buffer solution, 0.2mL of sodium azide solution (0.02g/mL) was added, followed by cellulase enzyme at 20 FPU/g-substrate and reaction at 50 ℃ for 72h at 120 rpm. Then, the glucose concentration in the enzymatic hydrolysate was measured by HPLC. The result shows that the concentration of glucose in the enzymolysis liquid is 21.6g/L, the concentration of xylose is 9.7g/L, and the final obtained total sugar yield is 75.1%.
Example 6
30.0 +/-0.1 g of bagasse is weighed, 300.0 +/-0.1 g of DES (choline chloride: lactic acid (1: 2)) is added for pretreatment for 1 hour at 90 ℃, solid-liquid separation is carried out after the reaction is finished, then 300.0 +/-0.1 mL of KOH solution with the mass fraction of 15 percent is added for pretreatment for 2 hours at 90 ℃, and solid-liquid separation is carried out after the reaction is finished. And (4) washing the solid part to be neutral by using tap water, and carrying out an enzymolysis saccharification experiment. The volume of the enzymatic hydrolysis reaction was 20mL, 1g of pretreated bagasse (absolute dry basis) was added at a solids loading of 5 wt.%, 0.05mol/L citric acid buffer solution was added to adjust the pH of the enzymatic hydrolysis to 4.8, 0.2mL of sodium azide solution (0.02g/mL) was added, and then cellulase was added at a rate of 5 FPU/g-substrate at 120rpm for 72h at 50 ℃. Then, the glucose concentration in the enzymatic hydrolysate was measured by HPLC. The result shows that the concentration of glucose in the enzymolysis liquid is 18.4g/L, the concentration of xylose is 7.5g/L, and the final obtained total sugar yield is 55.1%.
Example 7
30.0 +/-0.1 g of bagasse is weighed, 300.0 +/-0.1 g of DES (choline chloride: lactic acid (1: 2)) is added for pretreatment for 1 hour at 90 ℃, solid-liquid separation is carried out after the reaction is finished, then 300.0 +/-0.1 mL of KOH solution with the mass fraction of 15 percent is added for pretreatment for 2 hours at 90 ℃, and solid-liquid separation is carried out after the reaction is finished. And (4) washing the solid part to be neutral by using tap water, and carrying out an enzymolysis saccharification experiment. The enzymatic hydrolysis reaction volume was 20mL, 0.4g of pretreated bagasse (absolute dry basis) was added at a 2 wt.% solids loading, 0.05mol/L citric acid buffer solution was added to adjust the pH of the enzymatic hydrolysis to 4.8, 0.2mL of sodium azide solution (0.02g/mL) was added, followed by cellulase enzyme at 20 FPU/g-substrate, and the reaction was carried out at 50 ℃ for 72h at 120 rpm. Then, the glucose concentration in the enzymatic hydrolysate was measured by HPLC. The result shows that the concentration of glucose in the enzymolysis liquid is 9.7g/L, the concentration of xylose is 4.3g/L, and the final obtained total sugar yield is 74.2%.
Example 8
30.0 +/-0.1 g of bagasse is weighed, 300.0 +/-0.1 g of DES (choline chloride: lactic acid (1: 2)) is added for pretreatment for 1 hour at 90 ℃, solid-liquid separation is carried out after the reaction is finished, then 300.0 +/-0.1 mL of KOH solution with the mass fraction of 15 percent is added for pretreatment for 2 hours at 90 ℃, and solid-liquid separation is carried out after the reaction is finished. And (4) washing the solid part to be neutral by using tap water, and carrying out an enzymolysis saccharification experiment. The enzymatic hydrolysis reaction volume was 20mL, 2g of pretreated bagasse (absolute dry basis) was added at 10 wt.% solids loading, the pH of the enzymatic hydrolysis was adjusted to 4.8 by the addition of 0.05mol/L citric acid buffer solution, 0.2mL of sodium azide solution (0.02g/mL) was added, followed by cellulase enzyme at 20 FPU/g-substrate and reaction at 50 ℃ for 72h at 120 rpm. Then, the glucose concentration in the enzymatic hydrolysate was measured by HPLC. The result shows that the concentration of glucose in the enzymolysis liquid is 47.6g/L, the concentration of xylose is 21.6g/L, and the final obtained total sugar yield is 75.6%.
Example 9
Example 9 the same procedure as in example 2, except that: by adopting KOH solution with the mass fraction of 20 percent, the yield of the obtained glucose is 76.2 percent.
Example 10
Example 10 is the same as example 2, except that: by adopting KOH solution with the mass fraction of 40 percent, the yield of the obtained glucose is 72.8 percent.
Example 11
Example 11 is the same as example 2, except that: DES consists of choline chloride: lactic acid was added in a molar ratio of 1:1, preparing; the time of DES pretreatment of bagasse is 6h, the pretreatment solid-to-liquid ratio is 1:8g/mL, and the pretreatment temperature is 60 ℃; the pretreatment conditions of the KOH solution are as follows: the treatment time is 30min, the treatment solid-to-liquid ratio is 1:6g/mL, and the treatment temperature is 100 ℃; the enzymatic hydrolysis reaction volume was 20mL, with 4g of pretreated bagasse (absolute dry) added at a solids loading of 20 wt.%; cellulase was added at 3 FPU/g-substrate, solids loading at 2 wt.%, and enzymatic hydrolysis at pH 4.8 and temperature 50 ℃ for 2 h.
Example 12
Example 12 the same procedure as in example 2, except that: DES consists of choline chloride: lactic acid was added in a molar ratio of 1: 5, preparing the product; the time of DES pretreatment of bagasse is 0.5h, the pretreatment solid-to-liquid ratio is 1:30g/mL, and the pretreatment temperature is 130 ℃; the pretreatment conditions of the KOH solution are as follows: the treatment time is 180min, the treatment solid-to-liquid ratio is 1:18g/mL, and the treatment temperature is 50 ℃; cellulase was added at 30 FPU/g-substrate, solid loading was 20 wt.%, and enzymatic hydrolysis was carried out at pH 4.8 and temperature 50 ℃ for 96 h.
Example 13
Example 13 is a comparative example, and example 13 is prepared identically to example 2, except that: the enzymatic hydrolysis reaction volume was 20mL, 1g of untreated bagasse raw material (in absolute dry basis) was added at a solids loading of 5 wt.%, 0.05mol/L citric acid buffer solution was added to adjust the pH to 4.8, 0.2mL of sodium azide solution (0.02g/mL) was added, followed by cellulase enzyme at 20 FPU/g-substrate, and the reaction was carried out at 50 ℃ for 72h at 120 rpm. Then, the glucose concentration in the enzymatic hydrolysate was measured by HPLC. The results show that the concentration of glucose in the enzymolysis liquid is 3.4g/L, the concentration of xylose in the enzymolysis liquid is 1.0g/L, and the final obtained total sugar yield is 14.4 percent.
Comparing example 2 with optimal conditions it can be seen that example 13 is the same as example 2 in terms of enzymatic hydrolysis of the same bagasse feedstock, but example 13 directly performs enzymatic saccharification on the non-pretreated bagasse feedstock, whereas example 2 pre-treats the bagasse before enzymatic hydrolysis with DES and 15 wt.% KOH at 90 ℃ for 2h with a solid to liquid ratio of 1:10, the final enzyme hydrolysate has a glucose concentration of 21g/L higher than example 13, a xylose concentration of 10g/L higher than example 2, and the final total sugar yield of example 2 is 66.2% higher than example 13.
Observing fig. 1, fig. 2 and fig. 3, it can be seen that the bagasse raw material after pretreatment has increased crystallinity and increased porosity and porosity compared to the bagasse raw material without pretreatment, thereby increasing the accessibility of cellulase to cellulose and improving the efficiency of enzymatic saccharification. Example 2 is the optimum treatment condition of the present invention, examples 1-2 are the KOH dosage is gradually increased, the same enzyme hydrolysis condition is adopted to prove the influence of the KOH dosage on the sugar yield and the sugar concentration, the mass fraction of the KOH solution is 5-30 wt.% and has good effect, but the cost is consumed when the KOH dosage is too large, the over hydrolysis is caused when the dosage is too large, and the pretreatment effect of 15 wt.% KOH is the best; examples 2-4 compare the effect of temperature on sugar yield and sugar concentration, with KOH solution pretreatment at 90 ℃ for 2h giving the highest sugar yield; examples 2 and 5 compare the effect of different times on sugar yield and sugar concentration, and examples 2 and 6 compare the effect of different enzyme dosages on sugar yield and sugar concentration; examples 2, 7, 8 compare the effect of enzymatic hydrolysis on sugar yield and sugar concentration at different solids contents.
The method adopts DES and KOH combined treatment, can remove lignin to improve the sugar yield after enzymolysis and saccharification by two methods, but the removal rate of the lignin is not high and the effect is not obvious when the DES is carried out independently, the effect of the KOH pretreatment is when the DES is carried out independently, the using amount of the KOH needed is larger and the reaction condition is violent, the two methods are combined to have obvious synergistic effect, the DES is a green solvent, parts of lignin are removed by the DES treatment, and then the KOH treatment is added, so that the using amount of the KOH can be reduced to achieve the same effect.
Example 14
The liquid fraction collected by the solid-liquid separation in steps (1) and (2) of example 4 was used as a raw material for the preparation of a potassium fertilizer, a lignin-based fertilizer or a compound fertilizer.

Claims (8)

1, method for improving enzymolysis saccharification effect by bagasse pretreatment, which is characterized by comprising the following steps:
(1) weighing bagasse, adding DES (data encryption standard) for pretreatment, performing solid-liquid separation after the reaction is finished, and respectively collecting solid and liquid parts;
(2) adding KOH solution into the solid separated in the step (1) for pretreatment, carrying out solid-liquid separation on the solid after the reaction is finished, and respectively collecting the solid part and the liquid part;
(3) and (3) washing the solid part separated in the step (2) to be neutral to obtain pretreated bagasse, carrying out enzymolysis saccharification, and determining the concentration of glucose in the enzymolysis liquid.
2. The method for improving enzymolysis saccharification effect by bagasse pretreatment according to claim 1, wherein the DES is prepared by mixing choline chloride: lactic acid is preferably used in a molar ratio of 1: 1-5.
3. The method for improving the enzymolysis saccharification effect by bagasse pretreatment according to claim 1, wherein the conditions of the DES pretreatment of bagasse are as follows: the pretreatment time is 0.5-6h, the pretreatment solid-to-liquid ratio is 1:8-1:30g/mL, and the pretreatment temperature is 60-130 ℃.
4. The method for improving enzymatic saccharification effect by bagasse pretreatment according to claim 1, wherein the mass fraction of the KOH solution is 5-40 wt.%.
5. The method for improving enzymatic saccharification effect by bagasse pretreatment according to claim 1, wherein the pretreatment conditions of the KOH solution are as follows: the treatment time is 0.5-5h, the treatment solid-liquid ratio is 1:6-1:15g/mL, and the treatment temperature is 50-100 ℃.
6. The method for improving enzymatic saccharification effect through bagasse pretreatment according to claim 1, wherein the enzymatic saccharification is implemented by adding citric acid buffer solution to the pretreated bagasse to adjust the pH value, adding sodium azide solution, then adding cellulase, and measuring the glucose concentration in the enzymatic hydrolysate by using HPLC after reaction.
7. The method for improving enzymatic saccharification effect by bagasse pretreatment according to claim 6, wherein the cellulase is added in an amount of 3-30 FPU/g-substrate, the solid loading is 2-20 wt.%, and the enzymatic hydrolysis is performed at pH 4.8 and temperature of 50 ℃ for 2-96 h.
Use of liquid produced in the method for improving enzymatic saccharification effect by bagasse pretreatment as described in claim 1 for the production of potash fertilizer, lignin-based fertilizer and compound fertilizer.
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CN111763695A (en) * 2020-07-28 2020-10-13 暨南大学 Method for preparing ethanol fuel by pretreating bagasse with sodium phosphate
CN113603899A (en) * 2021-07-12 2021-11-05 华南理工大学 Method for pretreating lignocellulose by using Lewis base to assist neutral eutectic solvent
CN113882176A (en) * 2021-09-17 2022-01-04 华南理工大学 Method for preparing chemi-biomechanical pulp by promoting DES solution and bagasse treatment by biological enzyme by using biosurfactant
CN114045315A (en) * 2021-11-16 2022-02-15 中国科学院广州能源研究所 Method for improving enzymolysis efficiency of licorice residue

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111763695A (en) * 2020-07-28 2020-10-13 暨南大学 Method for preparing ethanol fuel by pretreating bagasse with sodium phosphate
CN113603899A (en) * 2021-07-12 2021-11-05 华南理工大学 Method for pretreating lignocellulose by using Lewis base to assist neutral eutectic solvent
CN113882176A (en) * 2021-09-17 2022-01-04 华南理工大学 Method for preparing chemi-biomechanical pulp by promoting DES solution and bagasse treatment by biological enzyme by using biosurfactant
CN113882176B (en) * 2021-09-17 2022-12-13 华南理工大学 Method for preparing chemi-bio-mechanical pulp
CN114045315A (en) * 2021-11-16 2022-02-15 中国科学院广州能源研究所 Method for improving enzymolysis efficiency of licorice residue

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