CN114540430A - Processing method of empty palm fruit clusters - Google Patents
Processing method of empty palm fruit clusters Download PDFInfo
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/16—Butanols
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- C12P19/00—Preparation of compounds containing saccharide radicals
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- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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- C12P2201/00—Pretreatment of cellulosic or lignocellulosic material for subsequent enzymatic treatment or hydrolysis
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P2203/00—Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source
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- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/30—Fuel from waste, e.g. synthetic alcohol or diesel
Abstract
The invention provides a processing method of empty palm fruit clusters, which comprises the following steps: mixing the empty palm fruit clusters with a mixed acid solution, and pretreating to obtain a first mixture; and adjusting the pH value of the first mixture, mixing the first mixture with cellulase, and performing enzymolysis to obtain a mixed hydrolysate. Therefore, the palm empty fruit clusters are treated by the mixed acid solution, the saccharification efficiency can be improved, meanwhile, the corrosion to equipment is reduced, enzymolysis is directly carried out after the mixed acid treatment, a large amount of water is not needed for cleaning, the water consumption is saved, the treatment cost is reduced, and the utilization rate of the palm empty fruit clusters is improved.
Description
Technical Field
The invention relates to the technical field of biochemical engineering, in particular to a method for processing empty palm fruit clusters.
Background
The palm Empty Fruit Bunch (EFB) is not only the most major waste in the palm oil industry, but also a renewable biomass resource abundant in nature. At present, only a small amount of EFB is used for preparing fibers and fertilizers, and most of EFB is directly burnt, so that the environment is polluted, and the resource waste is caused. The EFB contains a large amount of cellulose (35-50%), hemicellulose (15-30%) and lignin (10-20%), wherein the cellulose and the hemicellulose can be converted into reducing sugar under certain conditions and used as a carbon source for microbial fermentation. Therefore, EFB can be used as a cheap raw material to prepare high value-added products through microbial fermentation, for example, EFB is used as a raw material to obtain cellulose and hemicellulose hydrolysate through pretreatment, biological butanol is produced through microbial fermentation, and then biological aviation kerosene is further prepared through catalytic conversion. Due to the complex structure of EFB, wherein hemicellulose and lignin are intertwined to form a complex, cellulose is wrapped to form a complex compact structure, and the hydrolysis of cellulase is hindered. The commonly used pretreatment methods include dilute acid method, steam explosion method, ammonia fiber explosion, alkali method, biological method, organic solution method and the like, but most pretreatment methods are not beneficial to large-scale application due to higher cost or large wastewater discharge amount, wherein the dilute acid pretreatment cost is relatively lower, and the method is considered to have the most industrialized prospect at present. However, the conventional dilute acid pretreatment has many disadvantages: taking sulfuric acid as an example, the corrosion to equipment is serious; the low solid-to-liquid ratio results in large use amounts of acid and water; the hydrolysate has more inhibitors and high salt ion concentration, and has larger influence on the subsequent microbial fermentation. The invention provides a method for processing palm empty fruit clusters based on EFB as a raw material to produce biological aviation kerosene, which aims to solve the problems and provide a theoretical basis for the biological butanol production through fermentation after the EFB is pretreated and further the biological aviation kerosene is prepared.
Disclosure of Invention
The present invention is directed to solving, at least to some extent, one of the technical problems in the related art. Therefore, the invention aims to provide a pretreatment method of empty palm fruit bunches, which improves the saccharification efficiency, reduces the corrosion to equipment by using a small amount of acid, does not need to use a large amount of water in the treatment process, and has low treatment cost.
In one aspect of the invention, a processing method of empty palm fruit clusters is provided, which comprises the following steps: mixing empty palm fruit clusters with a mixed acid solution, and pretreating to obtain a first mixture; and adjusting the pH value of the first mixture, mixing the first mixture with cellulase, and carrying out enzymolysis to obtain a mixed hydrolysate. Therefore, the palm empty fruit clusters are treated by the mixed acid solution to replace sulfuric acid, the saccharification efficiency can be improved, meanwhile, corrosion to equipment is reduced, enzymolysis is directly carried out after the mixed acid treatment, a large amount of water is not needed for cleaning, the water consumption is saved, and the treatment cost is reduced.
In another aspect of the present invention, a method for processing empty palm fruit clusters is provided, which comprises the following steps: mixing empty palm fruit clusters with a mixed acid solution, and pretreating to obtain a first mixture; filtering the first mixture to obtain hemicellulose hydrolysate and filter residue; mixing the filter residue with water to obtain a second mixture, adjusting the pH value of the second mixture, adding cellulase for enzymolysis, and filtering to obtain cellulose hydrolysate. Therefore, the empty palm fruit clusters are treated by the mixed acid solution instead of sulfuric acid, so that the saccharification efficiency can be improved, and the corrosion to equipment is reduced.
According to some embodiments of the invention, the temperature of the pretreatment is 130 to 170 ℃, and the time of the pretreatment is 30 to 60 min.
According to some embodiments of the invention, the mass ratio of the empty palm fruit clusters to the mixed acid solution is 1 (2-6).
According to some embodiments of the invention, the mixed acid solution comprises sulfuric acid and phosphoric acid.
According to some embodiments of the invention, the mass ratio of the sulfuric acid to the phosphoric acid is 1: (0.5 to 1.5).
According to some embodiments of the invention, the concentration of the mixed acid in the mixed acid solution is 0.5% to 2% by mass.
According to some embodiments of the invention, the pH of the first mixture after pH adjustment and the second mixture after pH adjustment are each independently 4.8 to 5.2.
According to some embodiments of the invention, the cellulase is used in an amount of 15 to 25FPU per gram of empty palm fruit bunch.
According to some embodiments of the invention, the mass ratio of the filter residue to the water is 1: 4-1: 6.
According to some embodiments of the invention, the temperature of the enzymolysis is 45-50 ℃, the rotation speed is 130-160 rpm, and the time is 68-75 h.
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The above and/or additional aspects and advantages of the present invention will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:
FIG. 1 shows a schematic diagram of a method for pre-processing empty palm fruit bunches according to an embodiment of the present invention;
fig. 2 shows a schematic diagram of a method for pre-processing empty palm fruit clusters according to another embodiment of the invention.
Detailed Description
The following describes embodiments of the present invention in detail. The following examples are illustrative only and are not to be construed as limiting the invention. The examples, where specific techniques or conditions are not indicated, are to be construed according to the techniques or conditions described in the literature in the art or according to the product specifications. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.
In one aspect of the invention, a processing method of empty palm fruit clusters is provided, which comprises the following steps: mixing empty palm fruit clusters with a mixed acid solution, and pretreating to obtain a first mixture; and adjusting the pH value of the first mixture, mixing the first mixture with cellulase, and performing enzymolysis to obtain a mixed hydrolysate. Therefore, the palm empty fruit clusters are treated by using the mixed acid solution, the saccharification efficiency can be improved, meanwhile, the corrosion to equipment is reduced, enzymolysis is directly carried out after the mixed acid treatment, a large amount of water is not needed for cleaning, the water consumption is saved, the treatment cost is reduced, and the utilization rate of the palm empty fruit clusters is improved.
For convenience of understanding, the following detailed description is provided for the principle that the method can achieve the above beneficial effects:
as mentioned above, the treatment of the empty palm fruit bunch at present mainly uses sulfuric acid, which has serious corrosion to equipment, short service life of equipment, low solid-liquid ratio and large using amount of acid and water, and the sulfuric acid is used for pretreatment, so that a plurality of byproducts are generated in the hydrolysis process, and if the hydrolysate is used for microbial fermentation, the microbial fermentation process can be inhibited, and the sulfuric acid is used for pretreatment, so that the hydrolysate has strong acidity, when the hydrolysate is used for microbial fermentation, sodium hydroxide is needed to adjust the pH value of the hydrolysate to be neutral, the needed concentration of salt ions is high, and the subsequent microbial fermentation is greatly influenced. The method has the advantages that the crystallinity of cellulose is reduced, hemicellulose in the pretreated first mixture is hydrolyzed into reducing sugar, and after the cellulase is added into the first mixture, the cellulase can be in direct contact with the cellulose, the accessibility of the cellulase in the enzymolysis process is increased, the enzymolysis efficiency is improved, the saccharification efficiency is improved, the empty palm fruit clusters are converted into the reducing sugar, the empty palm fruit clusters can be used as a carbon source for microbial fermentation, and the utilization rate of the empty palm fruit clusters is improved.
In the following, the steps of the method are explained in detail according to an embodiment of the invention, and with reference to fig. 1, the method comprises:
s100: pretreatment of
In this step, the palm empty fruit bunches are mixed with a mixed acid solution, pre-treated to obtain a first mixture.
According to some embodiments of the invention, in order to make the empty palm fruit bunches and the mixed acid fully contact, the empty palm fruit bunches may be sieved before being mixed with the mixed acid, and the particle size distribution of the processed empty palm fruit bunches is more uniform and the processed empty palm fruit bunches and the mixed acid are more uniform, which is beneficial for hydrolysis. Specifically, the mesh number of the sieve is not particularly limited, and those skilled in the art can select the mesh number according to the original state of the empty palm fruit bunch.
According to some embodiments of the present invention, the kind of the palm empty fruit bunch is not particularly limited, and may be, for example, a palm Empty Fruit Bunch (EFB), a Wet palm empty fruit bunch (Wet-EFB), and a Deoiled palm empty fruit bunch (Deoiled-FEB).
According to some embodiments of the present invention, the mixing manner of the empty palm fruit bunch and the mixed acid is not particularly limited, for example, the acid components in the mixed acid may be sequentially added to the empty palm fruit bunch, and in order to mix the empty palm fruit bunch and the various acids in the mixed acid uniformly, the various acid components may be mixed first, and after mixing uniformly, the empty palm fruit bunch may be added to the mixed acid, or the mixed acid may be added to the empty palm fruit bunch.
According to some embodiments of the present invention, the mass ratio of the palm empty fruit bunches to the mixed acid solution is not particularly limited as long as hemicellulose in the palm empty fruit bunches can be hydrolyzed. Specifically, the mass ratio of the palm empty fruit bunch to the mixed acid solution is 1 (1-6), specifically, 1:2, 1:3, 1:4 and 1:5, preferably, the mass ratio of the empty palm fruit clusters to the mixed acid solution is 1 (4-5), so that the hydrolysis efficiency of the hemicellulose can be improved, the generation of hydrolysis inhibitors can be reduced, and the hemicellulose hydrolysate can be used for subsequent microbial fermentation. The inventor finds that if the mass ratio of the empty palm fruit clusters to the mixed acid solution is too large, the hydrolysis effect on hemicellulose is poor; if the mass ratio of the oil palm empty fruit clusters to the mixed acid solution is too small, although the hydrolysis efficiency of hemicellulose can be improved to a certain extent, the generated hydrolysis inhibitors are more, if the oil palm empty fruit clusters are used for microbial fermentation, the fermentation is not facilitated, and the water resource is wasted due to the fact that the water consumption is too large.
According to some embodiments of the present invention, the content of the mixed acid in the mixed acid solution is not particularly limited, and those skilled in the art can select the mixed acid according to the type of the acid, the performance of the empty palm fruit bunch, and the specific process of hydrolysis, and specifically, according to the present invention, the mass percentage concentration of the mixed acid in the mixed acid solution is 0.5% to 2%, so that the hydrolysis efficiency of hemicellulose can be improved, and at the same time, the equipment cannot be corroded. The inventor finds that if the content of the mixed acid in the mixed acid solution is too low, the hydrolysis efficiency of the hemicellulose is low, the concentration of reducing sugar in the hydrolysate is low, namely the saccharification rate is low; if the content of the mixed acid in the mixed acid solution is too high, hydrolysis inhibitors are large, and the equipment is corroded, so that the service life of the equipment is shortened.
According to some embodiments of the invention, the type of the mixed acid is not particularly limited, and when the mixed hydrolysate is used for microbial fermentation, the mixed acid can be selected from sulfuric acid and phosphoric acid, so that not only can the hydrolysis effect be ensured, but also phosphate radicals in the mixed hydrolysate can be used as a phosphorus source for subsequent microbial fermentation, and phosphorus does not need to be supplemented separately in the microbial fermentation process, thereby saving the cost to a certain extent.
According to some embodiments of the present invention, when sulfuric acid and phosphoric acid are included in the mixed acid, the mass ratio of sulfuric acid to phosphoric acid is 1: (0.5 to 1.5), whereby the saccharification rate is high, the corrosion to equipment is small, and the hydrolysis inhibitor is small in the obtained mixed hydrolysate. The inventor finds that the hydrolysis effect of sulfuric acid is better in the hydrolysis process of the empty palm fruit clusters, but if the sulfuric acid is too much, the content of phosphoric acid is too little, the phosphorus content in the final mixed hydrolysate is less, the requirement of microbial fermentation on phosphorus cannot be met, and the hydrolysis inhibitor is more and the equipment corrosion is higher due to too much sulfuric acid; if the phosphoric acid is too much and the content of sulfuric acid is low, more mixed acid solution needs to be added into the empty palm fruit strings in order to achieve the same hydrolysis effect.
According to some embodiments of the present invention, in order to enable the hydrolysis process to be performed more completely, after mixing the empty palm fruit clusters with a certain proportion of mixed acid solution, the empty palm fruit clusters may be placed in a pretreatment device for pretreatment for 30 to 60min, specifically, for 40min, 45min, 50min, 55min, and the like, and further, the first mixture may be heated during the pretreatment process, and the heating temperature may be set to 130 to 170 ℃, specifically, for 140 ℃, 145 ℃, 150 ℃, 155 ℃, 160 ℃, 165 ℃, and the like, so that the saccharification rate is high and the hydrolysis inhibitor is less. The inventors found that if the heating temperature is low, the hydrolysis efficiency is low and the saccharification rate is low; if the heating temperature is too high, hemicellulose is converted into aldehydes, and hydrolysis inhibitors are increased. It should be noted that the pretreatment device referred to herein may be a reaction vessel or an autoclave.
S200: enzymolysis
In this step, the pH of the first mixture is adjusted, and the first mixture is mixed with cellulase enzymes for enzymatic hydrolysis to obtain a mixed hydrolysate. According to some embodiments of the present invention, after the pretreatment with the mixed acid, the hemicellulose in the first mixture is hydrolyzed into reducing sugars, and in order to hydrolyze the cellulose in the first mixture into reducing sugars, cellulase is added to the first mixture for enzymolysis to obtain a mixed hydrolysate, wherein the mixed hydrolysate includes various reducing sugars that can be used as a carbon source for subsequent microbial fermentation.
According to some embodiments of the present invention, in order to make the enzymatic hydrolysis efficiency of the cellulase higher, the pH of the first mixture may be adjusted before adding the cellulase, specifically, the pH of the first mixture may be adjusted to 4.8 to 5.2, and when the pH is in this range, the enzymatic hydrolysis efficiency of the cellulase is highest, and the cellulose in the first mixture may be rapidly hydrolyzed into reducing sugar. Here, the type of the pH adjuster is not particularly limited, and may be, for example, calcium carbonate.
According to some embodiments of the invention, the cellulase can be added according to the amount of the empty palm fruit clusters, and particularly, according to the invention, the cellulase is used in an amount of 15-25 FPU/g of the empty palm fruit clusters, so that the cellulase can hydrolyze cellulose in the empty palm fruit clusters, and the saccharification rate is improved. If the dosage of the cellulase is too low, the empty palm fruit clusters cannot be completely hydrolyzed, the saccharification rate is low, the content of the cellulase is too high, and after cellulose is hydrolyzed, the cost is increased by excessive cellulase, so that waste is caused.
According to some embodiments of the present invention, in order to improve the efficiency of the enzymatic hydrolysis reaction, after adding the cellulase, the enzymatic hydrolysis reaction can be performed more completely when the first mixture containing the cellulase is placed on a shaker, specifically, the temperature of the enzymatic hydrolysis can be set to 45 ℃ to 50 ℃, the rotation speed is 130rpm to 160rpm, the time is 68h to 75h, and after the reaction is finished, the mixture is filtered, and the obtained filtrate is the mixed hydrolysate.
In another aspect of the present invention, a method for pre-processing empty palm fruit clusters is provided, which comprises the following steps: mixing empty palm fruit clusters with a mixed acid solution, and pretreating to obtain a first mixture; filtering the first mixture to obtain hemicellulose hydrolysate and filter residue; mixing the filter residue with water to obtain a second mixture, adjusting the pH value of the second mixture, adding cellulase for enzymolysis, and filtering to obtain cellulose hydrolysate. Therefore, the empty palm fruit clusters are treated by the mixed acid solution instead of sulfuric acid, so that the saccharification efficiency can be improved, and the corrosion to equipment is reduced.
In the following, the steps of the method are explained in detail according to an embodiment of the invention, with reference to fig. 2, the method comprising:
s100: enzymolysis
In this step, the empty palm fruit clusters are mixed with the mixed acid solution for pretreatment to obtain a first mixture, which is the same as the pretreatment process described above and will not be described herein again.
S200: filtration
In this step, the first mixture is filtered to obtain a hemicellulose hydrolysate and a residue. Specifically, after pretreatment of the empty palm fruit bunch by the mixed acid solution, hemicellulose is hydrolyzed, and after filtration, the hydrolyzed reducing sugar exists in the filtrate, so that hemicellulose hydrolysate can be directly obtained, and unhydrolyzed cellulose exists in filter residue, and then the next step of treatment is carried out.
According to some embodiments of the invention, the filtration process can be cleaned by distilled water to clean the reducing sugar in the filter residue into the filtrate, and in order to dissolve the reducing sugar in the water as much as possible, the filtration process can be cleaned by distilled water at 60-85 ℃ to quickly clean the reducing sugar in the filter residue and save water. According to some embodiments of the present invention, suction filtration may be performed using a buchner funnel.
S300: enzymolysis
In the step, the filter residue is mixed with water to obtain a second mixture, the pH value of the second mixture is adjusted, cellulase is added for enzymolysis, and the cellulose hydrolysate is obtained after filtration.
According to some embodiments of the invention, the water may be distilled water.
According to some embodiments of the invention, the mass ratio of the filter residue to the water can be 1: 4-1: 6, the filter residue and the water are mixed more uniformly, and the enzymolysis efficiency is higher; the inventors found that if the amount of water is too small, even if water is added to the residue, the second mixture has poor fluidity and is not uniformly mixed, which is not favorable for the enzymatic hydrolysis; if the amount of water is too much, the concentration of sugar in the finally obtained hydrolysate is too low, and the requirement of the carbon source during microbial fermentation cannot be met.
According to some embodiments of the present invention, in order to make the enzymatic hydrolysis efficiency of the cellulase higher, the pH of the second mixture may be adjusted before adding the cellulase, specifically, the pH of the second mixture may be adjusted to 4.8 to 5.2, and when the pH is in this range, the enzymatic hydrolysis efficiency of the cellulase is highest, and the cellulose in the second mixture may be rapidly hydrolyzed into reducing sugar. Here, the pH adjustment method is not particularly limited, and for example, the second mixture may be washed with water to be neutral, and then the pH may be adjusted to 4.8 to 5.2 with hydrochloric acid, or the second mixture may be directly adjusted to pH 4.8 to 5.2 with calcium hydroxide after being mixed with water in a certain ratio without washing the second mixture to be neutral, and a person skilled in the art may select the method.
According to some embodiments of the present invention, the dosage of the cellulase and the enzymatic process are the same as the aforementioned enzymatic process, and are not described herein again.
Example 1
Weighing 20g of EFB into a 500mL conical flask, adding 80mL of mixed acid (1% sulfuric acid + 1% phosphoric acid) with the total acid concentration of 2%, fully mixing, placing in a pretreatment device, keeping at 140 ℃ for 30min, transferring the material in the conical flask into a Buchner funnel for suction filtration, washing with 20mL of distilled water at 80 ℃ to obtain filtrate which is hemicellulose hydrolysate, and drying solid residues in an oven at 50 ℃ for later use. The content of each component in the hydrolysate was determined by High Performance Liquid Chromatography (HPLC), and the concentration of each reducing sugar in the hydrolysate is shown in Table 1.
Example 2
Weighing 20g of Wet-EFB into a 500mL conical flask, adding 80mL of mixed acid (1% sulfuric acid + 1% phosphoric acid) with the total acid concentration of 2%, fully mixing, placing in a pretreatment device, keeping at 140 ℃ for 30min, transferring the material in the conical flask to a Buchner funnel for suction filtration, washing with 20mL of distilled water at 80 ℃ to obtain filtrate which is hemicellulose hydrolysate, and drying solid residues in an oven at 50 ℃ for later use. The content of each component in the hydrolysate was determined by High Performance Liquid Chromatography (HPLC), and the concentration of each reducing sugar in the hydrolysate is shown in Table 1.
Example 3
Weighing 20g of Deoiled-FEB into a 500mL conical flask, adding 80mL of mixed acid (1% sulfuric acid and 1% phosphoric acid) with the total acid concentration of 2%, fully mixing, placing in a pretreatment device, keeping at 140 ℃ for 30min, transferring the material in the conical flask to a Buchner funnel for suction filtration, washing with 20mL of distilled water at 80 ℃ to obtain filtrate which is hemicellulose hydrolysate, and drying solid residues in an oven at 50 ℃ for later use. The content of each component in the hydrolysate was determined by High Performance Liquid Chromatography (HPLC), and the concentration of each reducing sugar in the hydrolysate is shown in Table 1.
TABLE 1
Example 4
Weighing 1g of the solid residue in example 1 into a 50mL conical flask, adding 5mL of distilled water, adjusting the pH to 5 with hydrochloric acid, adding 0.2g of cellulase, placing the mixture in a shaking table at 45 ℃ and 150rpm for enzymolysis for 72h, filtering the obtained product, wherein the obtained filtrate is cellulose hydrolysate, and determining the content of each component in the hydrolysate by High Performance Liquid Chromatography (HPLC), wherein the concentration of each reducing sugar is shown in Table 2.
Example 5
Weighing 1g of the solid residue obtained in the example 2 into a 50mL conical flask, adding 5mL of distilled water, adjusting the pH to 5 with hydrochloric acid, adding 0.2g of cellulase, placing the mixture in a shaking table at 45 ℃ and 150rpm for enzymolysis for 72h, filtering the obtained product, wherein the obtained filtrate is cellulose hydrolysate, and determining the content of each component in the hydrolysate by using High Performance Liquid Chromatography (HPLC), wherein the concentration of each reducing sugar is shown in Table 2.
Example 6
Weighing 1g of the solid residue obtained in example 3 into a 50mL conical flask, adding 5mL of distilled water, adjusting the pH to 5 with hydrochloric acid, adding 0.2g of cellulase, placing the mixture in a shaking table for enzymolysis at 45 ℃ and 150rpm for 72 hours, filtering the obtained product, obtaining filtrate which is cellulose hydrolysate, and measuring the content of each component in the hydrolysate by using High Performance Liquid Chromatography (HPLC), wherein the concentration of each reducing sugar is shown in Table 2.
TABLE 2
Example 7
1g of EFB was weighed into a 50mL Erlenmeyer flask, and 4mL of a mixed acid (1% sulfuric acid + 1% phosphoric acid) having a total acid concentration of 2% was added thereto, mixed well, and then placed in a pretreatment apparatus, and kept at 140 ℃ for 30 min. After pretreatment, 0.1g of calcium carbonate, 1mL of distilled water and 0.2g of cellulase are added, after full mixing, the mixture is placed in a shaking table for enzymolysis for 72h at 45 ℃ and 150rpm, the obtained product is filtered, the obtained liquid is a mixed hydrolysate, the content of each component in the mixed hydrolysate is determined by High Performance Liquid Chromatography (HPLC), and the concentration of each reducing sugar is shown in Table 3.
Example 8
Weigh 1g of Wet-EFB into a 50mL Erlenmeyer flask, add 4mL of a 2% total acid mixed acid (1% sulfuric acid + 1% phosphoric acid), mix well and place in a pretreatment apparatus, and hold at 140 ℃ for 30 min. After pretreatment, 0.1g of calcium carbonate, 1mL of distilled water and 0.2g of cellulase are added, after full mixing, the mixture is placed in a shaking table for enzymolysis for 72h at 45 ℃ and 150rpm, the obtained product is filtered, the obtained liquid is a mixed hydrolysate, the content of each component in the mixed hydrolysate is determined by High Performance Liquid Chromatography (HPLC), and the concentration of each reducing sugar is shown in Table 3.
Example 9
1g of Deoiled-FEB was weighed into a 50mL Erlenmeyer flask, 4mL of a mixed acid (1% sulfuric acid + 1% phosphoric acid) with a total acid concentration of 2% was added, mixed well and placed in a pretreatment apparatus, and kept at 140 ℃ for 30 min. After pretreatment, 0.1g of calcium carbonate, 1mL of distilled water and 0.2g of cellulase are added, after full mixing, the mixture is placed in a shaking table for enzymolysis for 72h at 45 ℃ and 150rpm, the obtained product is filtered, the obtained liquid is a mixed hydrolysate, the content of each component in the mixed hydrolysate is determined by High Performance Liquid Chromatography (HPLC), and the concentration of each reducing sugar is shown in Table 3.
TABLE 3
Example 10
1g of EFB raw material was weighed into a 50mL Erlenmeyer flask, 4mL of a mixed acid (1% sulfuric acid + 1% phosphoric acid) having a total acid concentration of 2% was added, and after thorough mixing, the mixture was placed in a pretreatment apparatus and kept at 150 ℃ for 30 min. After pretreatment, 0.1g of calcium carbonate, 2mL of distilled water and 0.2g of cellulase are added, after full mixing, the mixture is placed in a shaking table for enzymolysis for 72h at 50 ℃ and 150rpm, the obtained product is filtered, the obtained liquid is a mixed hydrolysate, the content of each component in the hydrolysate is determined by High Performance Liquid Chromatography (HPLC), and the concentration of each reducing sugar is shown in table 4.
Example 11
Weigh 1g of Wet-EFB into a 50mL Erlenmeyer flask, add 4mL of mixed acid (1% sulfuric acid + 1% phosphoric acid) with a total acid concentration of 2%, mix well and place in a pretreatment apparatus, hold at 150 ℃ for 30 min. After pretreatment, 0.1g of calcium carbonate, 1mL of distilled water and 0.2g of cellulase are added, after full mixing, the mixture is placed in a shaking table for enzymolysis for 72h at 50 ℃ and 150rpm, the obtained product is filtered, the obtained liquid is a mixed hydrolysate, the content of each component in the hydrolysate is determined by High Performance Liquid Chromatography (HPLC), and the concentration of each reducing sugar is shown in table 4.
Example 12
1g of Deoiled-FEB was weighed into a 50mL Erlenmeyer flask, 4mL of a mixed acid (1% sulfuric acid + 1% phosphoric acid) having a total acid concentration of 2% was added, and after thorough mixing, the mixture was placed in a pretreatment apparatus and kept at 150 ℃ for 30 min. After pretreatment, 0.1g of calcium carbonate, 1mL of distilled water and 0.2g of cellulase are added, after full mixing, the mixture is placed in a shaking table for enzymolysis for 72h at 50 ℃ and 150rpm, the obtained product is filtered, the obtained liquid is a mixed hydrolysate, the content of each component in the hydrolysate is determined by High Performance Liquid Chromatography (HPLC), and the concentration of each reducing sugar is shown in table 4.
TABLE 4
Example 13
1g of EFB starting material was weighed into a 50mL Erlenmeyer flask, 3mL of a mixed acid (0.975% sulfuric acid + 0.975% phosphoric acid) with a total acid concentration of 1.95% was added, mixed well and placed in a pretreatment apparatus for 60min at 148 ℃. After pretreatment, 0.1g of calcium carbonate, 2mL of distilled water and 0.2g of cellulase are added, after full mixing, the mixture is placed in a shaking table for enzymolysis for 72h at 50 ℃ and 150rpm, the obtained product is filtered, the obtained liquid is a mixed hydrolysate, the content of each component in the hydrolysate is determined by High Performance Liquid Chromatography (HPLC), and the concentrations of glucose, xylose and arabinose are respectively 38.36g/L, 19.37g/L and 4.13 g/L.
As can be seen from examples 1 to 6, the empty palm fruit clusters are hydrolyzed by the mixed acid solution, and then cellulase is added to the mixture, so that the empty palm fruit clusters can be hydrolyzed into reducing sugar; as can be seen from examples 7 to 9, the empty palm fruit clusters are pretreated by the mixed acid solution to obtain a first mixture, and then the first mixture is filtered and subjected to enzymolysis to obtain a hemicellulose hydrolysate and a cellulose hydrolysate, which can also hydrolyze the empty palm fruit clusters into reducing sugars; examples 10 to 12 it can be seen that in order to increase the concentration of total sugars in the hydrolysate, the solid to liquid ratio during pretreatment was 1: preferably, the solid-to-liquid ratio in the pretreatment process is 1:5 when Wet-EFB and Deoiled-FEB are treated; examples 10 to 12 are compared with examples 1 to 9, and it can be seen that the higher the temperature in the pretreatment process, the higher the saccharification rate after mixing the empty palm fruit clusters with the mixed acid solution; example 13 is an optimal experimental parameter obtained by optimizing a plurality of experimental parameters such as acid concentration, solid-to-liquid ratio, pretreatment temperature, and the like in the palm empty fruit bunch treatment process, according to which the palm empty fruit bunch treatment has the minimum corrosion to equipment, the minimum water usage, the maximum saccharification efficiency, and the minimum hydrolysis inhibitor content.
In the description of the present invention, it is to be understood that the terms "central," "longitudinal," "lateral," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," "counterclockwise," "axial," "radial," "circumferential," and the like are used in the orientations and positional relationships indicated in the drawings for convenience in describing the invention and to simplify the description, and are not intended to indicate or imply that the referenced device or element must have a particular orientation, be constructed and operated in a particular orientation, and are not to be considered limiting of the invention.
Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless specifically defined otherwise.
In the present invention, unless otherwise expressly specified or limited, the terms "mounted," "connected," "secured," and the like are to be construed broadly, as for example: can be fixedly connected, can also be detachably connected or integrated; can be mechanically or electrically connected; they may be directly connected or indirectly connected through intervening media, or may be connected through the use of two elements or the interaction of two elements. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.
In the present invention, unless expressly stated or limited otherwise, the first feature "on" or "under" the second feature may be directly contacting the second feature or the first and second features may be indirectly contacting each other through intervening media. Also, a first feature "on," "over," and "above" a second feature may be directly or diagonally above the second feature, or may simply indicate that the first feature is at a higher level than the second feature. A first feature being "under," "below," and "beneath" a second feature may be directly under or obliquely under the first feature, or may simply mean that the first feature is at a lesser elevation than the second feature.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Moreover, various embodiments or examples and features of various embodiments or examples described in this specification can be combined and mixed by those skilled in the art without contradiction.
Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.
Claims (10)
1. A processing method of palm empty fruit clusters is characterized by comprising the following steps:
mixing the empty palm fruit clusters with a mixed acid solution, and pretreating to obtain a first mixture;
and adjusting the pH value of the first mixture, mixing the first mixture with cellulase, and carrying out enzymolysis to obtain a mixed hydrolysate.
2. A processing method of empty palm fruit clusters is characterized by comprising the following steps:
mixing the empty palm fruit clusters with a mixed acid solution, and pretreating to obtain a first mixture;
filtering the first mixture to obtain hemicellulose hydrolysate and filter residue;
and mixing the filter residue with water to obtain a second mixture, adjusting the pH value of the second mixture, adding cellulase for enzymolysis, and filtering to obtain cellulose hydrolysate.
3. The processing method of the palm empty fruit bunch according to claim 1 or 2, wherein the temperature of the pretreatment is 130-170 ℃, and the time of the pretreatment is 30-60 min.
4. The method for processing the empty palm fruit bunches according to claim 1 or 2, wherein the mass ratio of the empty palm fruit bunches to the mixed acid solution is 1 (2-6).
5. The method for processing palm empty fruit bunches according to claim 1 or 2, wherein said mixed acid solution comprises sulfuric acid and phosphoric acid;
optionally, the mass ratio of the sulfuric acid to the phosphoric acid is 1: (0.5 to 1.5).
6. The method for processing empty palm fruit bunches according to claim 5, wherein the mass percentage concentration of the mixed acid in the mixed acid solution is 0.5-2%.
7. The method for processing palm empty fruit bunches according to claim 1 or 2, wherein the pH of the first mixture after pH adjustment and the pH of the second mixture after pH adjustment are each independently 4.8 to 5.2.
8. The method for processing the empty palm fruit bunches according to claim 1 or 2, wherein the dosage of the cellulase is 15-25 FPU/g of the empty palm fruit bunches.
9. The processing method of the palm empty fruit bunch according to claim 2, wherein the mass ratio of the filter residue to the water is 1: 4-1: 6.
10. The processing method of the palm empty fruit bunch according to claim 1 or 2, wherein the enzymolysis temperature is 45-50 ℃, the rotation speed is 130-160 rpm, and the time is 68-75 h.
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| US4378432A (en) * | 1980-09-25 | 1983-03-29 | Institut Francais Du Petrole | Process for manufacturing sweetened liquors and derivatives thereof from cellulose-containing vegetable substrates |
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| WO2020004473A1 (en) * | 2018-06-28 | 2020-01-02 | 関西化学機械製作株式会社 | Method for producing cellulase agent, and method for producing saccharification/fermentation product in which said cellulase agent is used |
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| US4378432A (en) * | 1980-09-25 | 1983-03-29 | Institut Francais Du Petrole | Process for manufacturing sweetened liquors and derivatives thereof from cellulose-containing vegetable substrates |
| WO2012049531A1 (en) * | 2010-10-12 | 2012-04-19 | Артер Текнолоджи Лимитед | Process for producing sugars by enzymatic hydrolysis of pretreated efb oil palm waste |
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