CN115927510A - Method for improving bagasse enzymolysis efficiency - Google Patents
Method for improving bagasse enzymolysis efficiency Download PDFInfo
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- CN115927510A CN115927510A CN202211685645.4A CN202211685645A CN115927510A CN 115927510 A CN115927510 A CN 115927510A CN 202211685645 A CN202211685645 A CN 202211685645A CN 115927510 A CN115927510 A CN 115927510A
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- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
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Abstract
The invention provides a method for improving bagasse enzymolysis efficiency, which comprises the steps of firstly, uniformly mixing bagasse, an organic solution and a metal salt, then carrying out heat preservation treatment, and then sequentially adding pretreated bagasse obtained by separation, polyethylene glycol 8000 and cellulase into a buffer solution for enzymolysis; wherein the metal salt is CaCl 2 、MgCl 2 、MnCl 2 、ZnCl 2 、FeCl 2 、CuCl 2 、AlCl 3 、FeCl 3 One or more of them. According to the invention, after bagasse is pretreated by adopting a specific metal salt and organic solvent coupling mode, the specific surfactant polyethylene glycol 8000 is added in an enzymolysis stage, and the enzymolysis efficiency of the bagasse is improved by cooperation with the metal salt, so that the yield of glucose in 24h of the bagasse reaches 87.8%, and the biological resource utilization of the bagasse is greatly promoted.
Description
Technical Field
The invention belongs to the technical field of biomass conversion and utilization. More particularly relates to a method for improving the enzymolysis efficiency of bagasse.
Background
Cellulose energy plants have high biomass yield, have high cellulose and hemicellulose content, are environment-friendly, are one of the most promising biomass energy sources at present, and are renewable resources which cannot be compared with petroleum.
Bagasse is a fibrous substrate of sugarcane stalks after crushing and extracting sugarcane juice from sugarcane, and is a very potential biomass refining resource. Bagasse hydrolysis can yield hexoses and pentoses, which can be fermented to ethanol, which can be added to gasoline as a fuel. However, cellulose and hemicellulose in bagasse are tightly connected with lignin, and the structure is stable, so that the cellulose is difficult to be effectively separated, and further utilization of the cellulose is influenced. Therefore, when biomass resources are converted, the biomass resources need to be pretreated to destroy the compact structure of the biomass resources and remove lignin in the biomass resources, so that hemicellulose and cellulose can be smoothly hydrolyzed into monosaccharides such as xylose and glucose in the enzymolysis process, and then ethanol is produced by further fermentation, so that the cellulose conversion rate is improved.
The conventional common bagasse pretreatment methods are generally physical methods, chemical methods and biological methods, and the chemical methods are the most probably industrialized production and most studied methods, and mainly comprise acid pretreatment, alkali pretreatment and organic solvent pretreatment. The organic solvent pretreatment is not usually enough to degrade lignin and hemicellulose in the lignocellulose biomass, and an acidic or alkaline catalyst is added to remove most of hemicellulose and lignin, break the original compact structure, improve the accessibility of cellulase and facilitate the subsequent utilization of cellulose, for example, the prior art provides a method for improving the bagasse enzymolysis efficiency by metal salt pretreatment and an additive, but the method takes Tween 80 as an additive and has a limited effect on improving the enzymolysis rate.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a method for improving the enzymolysis efficiency of bagasse, which takes a specific surfactant, namely polyethylene glycol 8000, as an additive to assist specific types of metal salts, and can effectively improve the enzymolysis rate of the bagasse.
The above purpose of the invention is realized by the following technical scheme:
the invention provides a method for improving bagasse enzymolysis efficiency, which comprises the following steps:
s1, uniformly mixing bagasse, an organic solution and metal salt, performing heat preservation treatment, and separating to obtain pretreated bagasse;
s2, sequentially adding the pretreated bagasse obtained in the step S1, polyethylene glycol 8000 and cellulase into a buffer solution for enzymolysis;
wherein the metal salt is CaCl 2 、MgCl 2 、MnCl 2 、ZnCl 2 、FeCl 2 、CuCl 2 、AlCl 3 、FeCl 3 One or more of them.
The method has mild pretreatment conditions, hemicellulose and lignin in the wood fiber raw material are effectively removed through organic solvent pretreatment, the addition of the metal salt further promotes the removal of the hemicellulose and the lignin, the compact structure of the bagasse raw material is broken down, more cellulose is exposed, the accessibility of enzyme to the cellulose is increased, the combination of the two pretreatment means more effectively removes the hemicellulose and the lignin in the bagasse, the influence of the hemicellulose and the lignin on subsequent enzymolysis is reduced, the cellulose is effectively retained, and the enzymolysis efficiency is preliminarily improved; in addition, the addition of the surfactant can reduce ineffective adsorption of lignin on the cellulase, stabilize the enzyme activity of cellulose and increase the accessibility of the cellulase on the cellulose, thereby effectively improving the yield of glucose in a short time.
Preferably, the metal salt is CuCl 2 、AlCl 3 、FeCl 3 One or more of them.
Preferably, the bagasse in S1 is air-dried, thread-rolled and crushed bagasse. The quality of the bagasse and the pretreated bagasse is calculated by absolute dry weight.
Preferably, the mass-to-volume ratio of the bagasse in S1 to the organic solution is 1g:8 to 15mL, most preferably 1g:10mL.
Preferably, the organic solution of S1 is an ethanol solution.
Preferably, the concentration of the organic solution of S1 is 50-70% (v/v).
Preferably, the final concentration of the metal salt of S1 in the organic solution is 0.02 to 0.03mol/L, more preferably 0.025mol/L.
Preferably, the rotation speed of the heat preservation treatment of S1 is 200-300 rpm.
Preferably, the temperature of the heat preservation treatment of S1 is 160-180 ℃.
Preferably, the time of the heat preservation treatment in S1 is 10-30 min.
Preferably, the heat preservation treatment of S1 is carried out in a closed environment, such as a reaction kettle and the like.
Preferably, the separation of S1 is vacuum filtration or centrifugation.
Preferably, the separation of S1 is further washed, and more preferably washed to neutrality by using an ethanol solution and deionized water in sequence.
Further preferably, the concentration of the ethanol solution is 55% to 65% (v/v).
Preferably, the dosage ratio of the pretreated bagasse to the cellulase in S2 is 1g:10 to 20FPU, most preferably 1g:10FPU.
Preferably, the mass ratio of the pretreated bagasse in S2 to the polyethylene glycol 8000 is 1:0.01 to 0.06, most preferably 1:0.025.
preferably, the pH of the buffer solution of S2 is 4.5-5.0.
Preferably, the buffer solution of S2 is acetic acid-sodium acetate buffer solution.
Preferably, the mass-to-volume ratio of the pretreated bagasse to the buffer solution in S2 is 1g:40 to 100mL, most preferably 1g:50mL.
Preferably, the temperature of the enzymolysis of S2 is 40-55 ℃, and most preferably 50 ℃.
Preferably, the rotation speed of the enzymolysis of S2 is 100-250 rpm, and most preferably 150rpm.
Preferably, the enzymolysis time of S2 is 6-72 h, and most preferably 24h.
The invention has the following beneficial effects:
according to the invention, after bagasse is pretreated by adopting a specific metal salt and organic solvent coupling mode, the specific surfactant polyethylene glycol 8000 is added in an enzymolysis stage, and the enzymolysis efficiency of the bagasse is improved by cooperation with the metal salt, so that the yield of glucose in 24h of the bagasse reaches 87.8%, and the biological resource utilization of the bagasse is greatly promoted.
Detailed Description
The present invention is further illustrated by the following specific examples, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.
Unless otherwise indicated, reagents and materials used in the following examples are commercially available.
The bagasse raw materials used in the invention are all bagasse which is air-dried, thread-rolled and crushed to the grain size of less than 1mm, and contain 40.2% of cellulose, 21.5% of hemicellulose, 25.2% of lignin and other components.
Cellulase enzymes (Seiland generation) were purchased from Novoxil (China) Biotechnology Ltd.
Example 1 method for improving bagasse enzymolysis efficiency
S1, air-drying 15g of bagasse after being subjected to thread rolling and crushing, 150mL of 60% (v/v) ethanol solution and 0.00375mol of CuCl 2 Uniformly mixing in a reaction kettle, carrying out heat preservation treatment at 280rpm and 160 ℃ for 10min, sequentially washing with 60% (v/v) ethanol solution and deionized water to be neutral, and carrying out vacuum filtration to obtain pretreated bagasse;
s2, sequentially adding 2g of the pretreated bagasse obtained in the S1, 0.05g of polyethylene glycol 8000 and 20FPU of cellulase into 100mL of acetic acid-sodium acetate buffer solution with the pH value of 4.8, and carrying out enzymolysis for 72h at 50 ℃ and 150rpm.
Example 2 method for improving bagasse enzymolysis efficiency
S1, 15g of air-dried, thread-rolled and crushed bagasse, 120mL of 50% (v/v) ethanol solution and 0.0024mol of CuCl 2 Uniformly mixing in a reaction kettle, carrying out heat preservation treatment at 200rpm and 180 ℃ for 10min, sequentially washing with 65% (v/v) ethanol solution and deionized water to be neutral, and carrying out vacuum filtration to obtain pretreated bagasse;
s2, sequentially adding 2g of the pretreated bagasse obtained in the S1, 0.02g of polyethylene glycol 8000 and 20FPU of cellulase into 80mL of acetic acid-sodium acetate buffer solution with the pH value of 4.5, and carrying out enzymolysis for 72 hours at 40 ℃ and 250 rpm.
Example 3 method for improving bagasse enzymolysis efficiency
S1, 15g of air-dried, thread-rolled and crushed bagasse, 225mL of 70% (v/v) ethanol solution and 0.00675mol of CuCl 2 Uniformly mixing in a reaction kettle, carrying out heat preservation treatment at 300rpm and 160 ℃ for 30min, sequentially washing with 55% (v/v) ethanol solution and deionized water to be neutral, and carrying out vacuum filtration to obtain pretreated bagasse;
s2, sequentially adding 2g of the pretreated bagasse obtained in the S1, 0.12g of polyethylene glycol 8000 and 40FPU of cellulase into 200mL of acetic acid-sodium acetate buffer solution with the pH value of 5.0, and carrying out enzymolysis for 72 hours at 55 ℃ and 100 rpm.
Example 4 method for improving bagasse enzymolysis efficiency
The same as example 1 except that CuCl was added 2 Replacement with CaCl 2 。
Example 5 method for improving bagasse enzymolysis efficiency
The same as example 1 except that CuCl was added 2 Substitution to MgCl 2 。
Example 6 method for improving bagasse enzymolysis efficiency
The same as example 1, except that CuCl is added 2 Replacement by MnCl 2 。
Example 7 method for improving bagasse enzymolysis efficiency
The same as example 1, except that CuCl is added 2 Substituted by ZnCl 2 。
Embodiment 8 method for improving bagasse enzymolysis efficiency
The same as example 1 except that CuCl was added 2 Replacement by FeCl 2 。
Example 9 method for improving bagasse enzymolysis efficiency
The same as example 1, except that CuCl is added 2 Replaced by AlCl 3 。
Example 10 method for improving bagasse enzymolysis efficiency
The same as example 1 except that CuCl was added 2 Replacement with FeCl 3 。
Comparative example 1
Same as example 1 except thatIs prepared by mixing CuCl 2 Replacement was with LiCl.
Comparative example 2
The same as example 1, except that CuCl is added 2 NaCl was substituted.
Comparative example 3
The same as example 4 except that polyethylene glycol 8000 was replaced with tween 80.
Comparative example 4
The same as example 5 except that polyethylene glycol 8000 was replaced with tween 80.
Comparative example 5
The difference from example 4 is that no polyethylene glycol 8000 is added.
Comparative example 6
The difference from example 5 is that polyethylene glycol 8000 was not added.
Comparative example 7
The difference from example 6 is that no polyethylene glycol 8000 is added.
Comparative example 8
The difference from example 1 is that CuCl is not added 2 。
Experimental example 1
The glucose concentrations of examples 1 to 10 and comparative examples 1 to 8 were measured by high performance liquid chromatography at 24 hours and 72 hours of enzymolysis, respectively, and the yields were calculated, and the results are shown in table 1.
TABLE 1 glucose yield of examples 1 to 10 and comparative examples 1 to 8 (unit:%)
As can be seen from the table:
(1) The glucose yield of examples 1-10 is significantly better than that of comparative examples 1-8, which shows that the method of the invention can effectively improve the enzymolysis efficiency of bagasse.
(2) The glucose yield of examples 1-10 is significantly better than that of comparative examples 1-2, which indicates that the method of the present invention selects specific metal salts, effectively removes hemicellulose and lignin, effectively retains cellulose, and has little damage to cellulose, so that the high enzymolysis efficiency can be realized, while LiCl and NaCl are difficult to effectively remove hemicellulose and lignin.
(3) The glucose yield of example 4 is significantly better than that of comparative example 3, and the glucose yield of example 5 is significantly better than that of comparative example 4, indicating that the specific surfactant polyethylene glycol 8000 is selected by the method of the present invention to achieve such high enzymolysis efficiency.
(4) Compared with the comparative example 5, the 24h glucose yield of the example 4 is increased by 29.82%, and the 72h glucose yield is increased by 11.43%; compared with the comparative example 6, the 24h glucose yield of the example 5 is increased by 22.02 percent, and the 72h glucose yield is increased by 6.49 percent; compared with the comparative example 7, the 24h glucose yield of the example 6 is increased by 26.02%, and the 72h glucose yield is increased by 7.92%. It is shown that the method of the invention can realize the high enzymolysis efficiency only by adding the surfactant polyethylene glycol 8000.
(8) The glucose yield of example 1 is significantly better than that of comparative example 8, which shows that the pretreatment method of metal salt reinforced organic solvent effectively improves the enzymolysis efficiency of glucose in bagasse.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. A method for improving bagasse enzymolysis efficiency is characterized by comprising the following steps:
s1, uniformly mixing bagasse, an organic solution and metal salt, performing heat preservation treatment, and separating to obtain pretreated bagasse;
s2, sequentially adding the pretreated bagasse obtained in the step S1, polyethylene glycol 8000 and cellulase into a buffer solution for enzymolysis;
wherein the metal salt is CaCl 2 、MgCl 2 、MnCl 2 、ZnCl 2 、FeCl 2 、CuCl 2 、AlCl 3 、FeCl 3 One or more of them.
2. The method according to claim 1, wherein the organic solution S1 is an ethanol solution.
3. The method of claim 1, wherein the final concentration of the metal salt of S1 in the organic solution is 0.02 to 0.03mol/L.
4. The method according to claim 1, wherein the rotation speed of the heat preservation treatment of S1 is 200-300 rpm; the temperature is 160-180 ℃; the time is 10-30 min.
5. The method according to claim 1, wherein the dosage ratio of the pretreated bagasse to the cellulase in S2 is 1g:10 to 20FPU.
6. The method according to claim 1, wherein the mass ratio of the pretreated bagasse to polyethylene glycol 8000 in S2 is 1:0.01 to 0.06 percent.
7. The method according to claim 1, wherein the buffer solution S2 is acetic acid-sodium acetate buffer solution.
8. The method according to claim 1, wherein the mass-to-volume ratio of the pretreated bagasse to the buffer solution is 1g: 40-100 mL.
9. The method according to claim 1, wherein the temperature of the S2 enzymolysis is 40-55 ℃; the time is 6 to 72 hours.
10. The method of claim 1, wherein the metal salt is CuCl 2 、AlCl 3 、FeCl 3 One or more of them.
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