CN114807250A - Method for producing succinic acid by fermentation - Google Patents
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- CN114807250A CN114807250A CN202210644070.5A CN202210644070A CN114807250A CN 114807250 A CN114807250 A CN 114807250A CN 202210644070 A CN202210644070 A CN 202210644070A CN 114807250 A CN114807250 A CN 114807250A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/44—Polycarboxylic acids
- C12P7/46—Dicarboxylic acids having four or less carbon atoms, e.g. fumaric acid, maleic acid
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P2201/00—Pretreatment of cellulosic or lignocellulosic material for subsequent enzymatic treatment or hydrolysis
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
Abstract
The application provides a method for producing succinic acid by fermentation, and relates to the technical field of succinic acid preparation. A method for producing succinic acid by fermentation comprises the following steps: adding water into the Chinese medicinal residue, performing ultrasonic treatment and heating and decocting, and performing solid-liquid separation to obtain first residue; drying the first filter residue, mixing with ammonia water, soaking, sealing, heating, and performing solid-liquid separation to obtain a second filter residue; carrying out enzymolysis on the second filter residue, and carrying out solid-liquid separation to obtain an enzymolysis liquid; adjusting the pH value of the enzymolysis liquid, sterilizing and inoculating zymocyte, and fermenting to produce the succinic acid. According to the method, the traditional Chinese medicine residues are used as raw materials for producing the succinic acid through fermentation, a large amount of cellulose and starch are fully utilized, the effect of resource recycling is achieved, the traditional Chinese medicine residues are subjected to enzymolysis pretreatment, the contact area of the cellulose and cellulase molecules is increased, the enzymolysis rate and the fermentation effect are improved, and the concentration of the succinic acid in the final fermentation liquid reaches 78-86 g/L.
Description
Technical Field
The application relates to the technical field of succinic acid preparation, in particular to a method for producing succinic acid by fermentation.
Background
Succinic acid, also known as succinic acid, is an important organic chemical raw material and intermediate, and is mainly used in the pharmaceutical industry, the food industry, the chemical industry, analytical reagents, food iron fortifier, preparation of electroplating liquid medicine and PCB (printed Circuit Board) liquid medicine, cleaning additives and the like.
The expansion of the application range of succinic acid promotes the research of the production method of succinic acid, and the production method of succinic acid mainly comprises a chemical synthesis method, a biological conversion method and a microbial fermentation method at present. The traditional succinic acid production method is mainly characterized in that a petrochemical method is applied to produce from butane through cis-butene dianhydride, the existing chemical preparation succinic acid production process is relatively mature, but the defects of high energy consumption, high pollution, low yield and purity, dependence on petrochemical resources and the like exist, and the development potential of the succinic acid as a bulk chemical is inhibited.
Only succinic acid used in food industry is produced by fermentation method, the succinic acid produced by fermentation method uses renewable sugar source (such as glucose) and carbon dioxide as main raw materials, it opens up a new way for utilizing greenhouse gas carbon dioxide, and has low cost, low energy consumption, little pollution and environmental protection, and is a novel 'green chemical industry' production process with high efficiency and low consumption. In recent years, due to the dual pressure of petroleum crisis and environmental pollution, the microbial fermentation method for producing succinic acid has attracted much attention because of its advantages of saving a large amount of petroleum resources and reducing pollution caused by petrochemical processes.
Disclosure of Invention
The application aims to provide a method for producing succinic acid by fermentation, which has the advantage of high succinic acid yield.
The technical problem to be solved by the application is solved by adopting the following technical scheme.
The embodiment of the application provides a method for producing succinic acid by fermentation, which comprises the following steps:
adding water into the Chinese medicinal residue, performing ultrasonic treatment and heating and decocting, and performing solid-liquid separation to obtain first residue;
drying the first filter residue, mixing with ammonia water, soaking, sealing, heating, and performing solid-liquid separation to obtain a second filter residue;
carrying out enzymolysis on the second filter residue, and carrying out solid-liquid separation to obtain an enzymolysis liquid;
adjusting the pH of the enzymolysis liquid to be neutral, sterilizing, inoculating zymocyte, and fermenting to produce the succinic acid.
Compared with the prior art, the embodiment of the application has at least the following advantages or beneficial effects:
the method adopts the traditional Chinese medicine residues as a carbon source for producing succinic acid by fermentation, firstly carries out ultrasonic treatment and heating boiling on the traditional Chinese medicine residues, can destroy the structure of lignocellulose in the traditional Chinese medicine residues under ultrasonic and high temperature, breaks the connection of partial cellulose and hemicellulose, and destroys an inner layer structure containing a large amount of lignin; on the basis, ammonia water is added to fully soak the lignocellulose, the lignocellulose is sealed and heated, the ammonia water can effectively soak the lignocellulose in the environment of high-temperature alkali liquor, chemical bonding between the lignin and the hemicellulose is completely destroyed, the polymerization degree of the lignin and the hemicellulose is reduced, most of the lignin and the hemicellulose surrounding the cellulose are removed, a cellulose crystallization area is fully exposed, the contact area between the cellulose and cellulase molecules is increased, the subsequent enzymolysis part is facilitated to carry out full enzymolysis, the fermentation effect is improved, the conversion rate of succinic acid is improved, and the concentration of succinic acid in the final fermentation liquor reaches 78-86 g/L.
This application adopts aqueous ammonia solution as alkali liquor heat treatment's raw materials, can improve the enzymolysis effect on the one hand, and on the other hand aqueous ammonia easily removes, has reduced subsequent sewage treatment process.
The method adopts the Chinese medicine residues as the raw materials for producing the succinic acid by fermentation, makes full use of a large amount of cellulose and starch in the raw materials, and plays a role in resource recycling.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions of the embodiments of the present application will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict. The present application will be described in detail below with reference to specific examples.
A method for producing succinic acid by fermentation comprises the following steps:
adding water into the Chinese medicinal residue, performing ultrasonic treatment and heating and decocting, and performing solid-liquid separation to obtain first residue;
drying the first filter residue, mixing with ammonia water, soaking, sealing, heating, and performing solid-liquid separation to obtain a second filter residue;
carrying out enzymolysis on the second filter residue, and carrying out solid-liquid separation to obtain an enzymolysis liquid;
adjusting the pH of the enzymolysis liquid to be neutral, sterilizing, inoculating zymocyte, and fermenting to produce the succinic acid.
The method adopts the traditional Chinese medicine residues as a carbon source for producing succinic acid by fermentation, firstly carries out ultrasonic treatment and heating boiling on the traditional Chinese medicine residues, can destroy the structure of lignocellulose in the traditional Chinese medicine residues under ultrasonic and high temperature, breaks the connection of partial cellulose and hemicellulose, and destroys an inner layer structure containing a large amount of lignin; on the basis, ammonia water is added to fully soak the lignocellulose, the lignocellulose can be sealed and heated, the ammonia water can effectively soak the lignocellulose in the environment of high-temperature alkali liquor, chemical bonding between the lignin and the hemicellulose is completely destroyed, the polymerization degree of the lignin and the hemicellulose is reduced, most of the lignin and the hemicellulose surrounding the cellulose are removed, a cellulose crystallization area is fully exposed, the contact area between the cellulose and cellulase molecules is increased, the subsequent enzymolysis part is fully hydrolyzed, the fermentation effect is improved, the concentration of succinic acid in the final fermented product reaches 78-86 g/L, and the conversion rate is 90-95%.
This application adopts aqueous ammonia solution as alkali liquor heat treatment's raw materials, can improve the enzymolysis effect on the one hand, and on the other hand aqueous ammonia easily removes, has reduced subsequent sewage treatment process.
The method adopts the Chinese medicine residues as the raw materials for producing the succinic acid by fermentation, makes full use of a large amount of cellulose and starch in the raw materials, and plays a role in resource recycling.
In some embodiments of the present application, the mass ratio of the herb residue to water is 1: (8-12), the ultrasonic intensity is 5-8W/m 2 And the ultrasonic time is 10-30 min.
In some embodiments of the present application, the heating and boiling temperature is 130-150 ℃ and the time is 20-40 min.
In some embodiments of the present application, the first filter residue is dried at 50 to 70 ℃ for 8 to 15 min.
In some embodiments of the present application, a solid-liquid mass ratio of the dried first filter residue to ammonia water is 1: (6-10), wherein the concentration of the ammonia water is 25-35%.
In some embodiments of the present application, the soaking time is 1 to 3 hours, the sealing and heating temperature is 100 to 120 ℃, and the heating time is 10 to 20 hours.
In some embodiments of the present application, the step of performing enzymolysis specifically includes: and adjusting the pH value of the second filter residue to 5.5-6.6, adding cellulase, cellobiase and alpha-amylase, and carrying out enzymolysis for 16-28 h at 35-40 ℃.
In some embodiments of the present application, the cellulase is added in an amount of 15 to 25IU/g, cellobiase is added in an amount of 5 to 10IU/g, and alpha-amylase is added in an amount of 5 to 10 IU/g.
In some embodiments of the present application, the pH is adjusted to 6.5 to 7.8 by using magnesium carbonate, the sterilization temperature is 120 to 125 ℃, and the sterilization time is 20 to 25 min.
In some embodiments of the present application, the fermentation bacteria is actinobacillus succinogenes F3-21, the inoculation amount is 4-8%, and 100% CO is introduced during the fermentation process 2 The fermentation temperature is 35-38 ℃.
The features and properties of the present application are described in further detail below with reference to examples.
Example 1
A method for producing succinic acid by fermentation comprises the following steps:
drying the residue (containing equal mass of calculus bovis, rhizoma corydalis, radix Angelicae sinensis, pericarpium Citri Tangerinae, fructus crataegi, Atractylodis rhizoma and Raphani semen) in oven at 50 deg.C to constant weight, mixing 500g with 5kg purified water, adding into ultrasonic instrument, and stirring at 6W/m 2 Performing ultrasonic treatment for 20min at the intensity of (1), taking out, putting into a reaction kettle, heating to 135 ℃, decocting for 15min, cooling to room temperature, filtering, and taking filter residue for later use;
drying the decocted filter residue in a 60 ℃ oven for 10min, placing the dried filter residue in a reaction kettle, adding 10 times of ammonia water (the concentration is 25%) by mass, sealing and soaking for 2h, heating to 110 ℃, reacting for 15h, cooling to room temperature, opening the reaction kettle, filtering, and taking the filter residue for later use;
adjusting the pH of the filter residue after the alkali liquor reaction to 5.8 by using dilute sulfuric acid, adding cellulase, cellobiase and alpha-amylase (the adding proportions are respectively 20IU/g, 8IU/g and 7IU/g), placing the mixture in an environment at 37 ℃ for reacting for 24 hours, and filtering to obtain an enzymatic hydrolysate;
adjusting pH of the enzymatic hydrolysate to 7.2 with magnesium carbonate solid, adding 35mg/L vitamin C, sterilizing at 125 deg.C for 21min, inoculating 5% Actinobacillus succinogenes F3-21 at 37 deg.C, and introducing 100% CO continuously during fermentation 2 And carrying out anaerobic fermentation for 50h, and finally measuring that the concentration of the succinic acid in the fermentation product reaches 84.6 g/L.
Example 2
A method for producing succinic acid by fermentation comprises the following steps:
drying the residue (containing equal mass of calculus bovis, rhizoma corydalis, radix Angelicae sinensis, pericarpium Citri Tangerinae, fructus crataegi, Atractylodis rhizoma and Raphani semen) in oven at 55 deg.C to constant weight, mixing 500g with 6kg purified water, adding into ultrasonic instrument, and stirring at 8W/m 2 Ultrasonic treating for 20min, taking out, placing into a reaction kettle, heating to 135 deg.CDecocting for 20min, cooling to room temperature, filtering, and collecting the residue;
drying the decocted filter residue in a 55 ℃ oven for 15min, placing the dried filter residue in a reaction kettle, adding 6 times of ammonia water (the concentration is 35%) by mass, sealing and soaking for 2h, heating to 110 ℃, reacting for 20h, cooling to room temperature, opening the reaction kettle, filtering, and taking the filter residue for later use;
adjusting the pH of the filter residue after the alkali liquor reaction to 6.2 by using dilute sulfuric acid, adding cellulase, cellobiase and alpha-amylase (the adding proportions are 22IU/g, 5IU/g and 5IU/g respectively), placing the mixture in an environment at 40 ℃ for reaction for 22 hours, and filtering the mixture to obtain an enzymatic hydrolysate;
adjusting pH of the enzymatic hydrolysate to 7.0 with magnesium carbonate solid, adding 40mg/L vitamin C, sterilizing at 121 deg.C for 25min, inoculating 8% Actinobacillus succinogenes F3-21, fermenting at 38 deg.C, and continuously introducing 100% CO during fermentation 2 And carrying out anaerobic fermentation for 55h, and finally measuring that the concentration of the succinic acid in the fermentation product reaches 85.9 g/L.
Example 3
A method for producing succinic acid by fermentation comprises the following steps:
drying the residue (containing equal mass of calculus bovis, rhizoma corydalis, radix Angelicae sinensis, pericarpium Citri Tangerinae, fructus crataegi, Atractylodis rhizoma and Raphani semen) in oven at 55 deg.C to constant weight, mixing 500g with 4kg purified water, adding into ultrasonic instrument, and stirring at 5W/m 2 Performing ultrasonic treatment for 25min at the intensity of (1), taking out, putting into a reaction kettle, heating to 150 ℃, decocting for 15min, cooling to room temperature, filtering, and taking filter residue for later use;
drying the decocted filter residue in a 70 ℃ oven for 8min, placing the dried filter residue in a reaction kettle, adding 8 times of ammonia water (the concentration is 30%) by mass, sealing and soaking for 2.5h, heating to 120 ℃, reacting for 18h, cooling to room temperature, opening the reaction kettle, filtering, and taking the filter residue for later use;
adjusting the pH of the filter residue after the alkali liquor reaction to 6.6 by using dilute sulfuric acid, adding cellulase, cellobiase and alpha-amylase (the adding proportions are respectively 25IU/g, 7IU/g and 5IU/g), placing the mixture in an environment at 35 ℃ for reacting for 28 hours, and filtering to obtain an enzymatic hydrolysate;
adjusting pH of the enzymatic hydrolysate to 7.8 with magnesium carbonate solid, adding 40mg/L vitamin C, sterilizing at 124 deg.C for 22min, inoculating 4% Actinobacillus succinogenes F3-21, fermenting at 35 deg.C, and continuously introducing 100% CO during fermentation 2 And carrying out anaerobic fermentation for 60 hours, and finally measuring that the concentration of the succinic acid in the fermentation product reaches 84.6 g/L.
Example 4
A method for producing succinic acid by fermentation comprises the following steps:
drying the residue (containing equal mass of calculus bovis, rhizoma corydalis, radix Angelicae sinensis, pericarpium Citri Tangerinae, fructus crataegi, Atractylodis rhizoma and Raphani semen) in oven at 50 deg.C to constant weight, mixing 500g with 5.8kg purified water, adding into ultrasonic instrument, and stirring at 8W/m 2 Performing ultrasonic treatment for 25min at the intensity of (1), taking out, putting into a reaction kettle, heating to 135 ℃, decocting for 25min, cooling to room temperature, filtering, and taking filter residue for later use;
drying the decocted filter residue in a 65 ℃ oven for 10min, placing the dried filter residue in a reaction kettle, adding 7 times of ammonia water (the concentration is 35%) by mass, sealing and soaking for 1h, heating to 120 ℃, reacting for 20h, cooling to room temperature, opening the reaction kettle, filtering, and taking the filter residue for later use;
adjusting the pH of the filter residue after the alkali liquor reaction to 5.5 by using dilute sulfuric acid, adding cellulase, cellobiase and alpha-amylase (the adding proportions are 15IU/g, 6IU/g and 10IU/g respectively), placing the mixture in an environment at 40 ℃ for reaction for 20 hours, and filtering the mixture to obtain an enzymatic hydrolysate;
adjusting pH of the enzymatic hydrolysate to 7.0 with magnesium carbonate, adding 40mg/L vitamin C, sterilizing at 125 deg.C for 20min, inoculating 7% Actinobacillus succinogenes F3-21, fermenting at 38 deg.C, and introducing 100% CO continuously during fermentation 2 And carrying out anaerobic fermentation for 55h, and finally measuring that the concentration of the succinic acid in the fermentation product reaches 82.7 g/L.
Example 5
A method for producing succinic acid by fermentation comprises the following steps:
decocting the Chinese medicinal residues (the residue contains equal amount of calculus bovis, rhizoma corydalis, radix Angelicae sinensis, pericarpium Citri Tangerinae, fructus crataegi, Atractylodis rhizoma and Raphani semen)Drying in oven at 55 deg.C to constant weight, collecting 500g, adding 4.8kg purified water, mixing, placing into ultrasonic instrument, and drying at 8W/m 2 Performing ultrasonic treatment for 30min at the intensity of (1), taking out, putting into a reaction kettle, heating to 130 ℃, decocting for 25min, cooling to room temperature, filtering, and taking filter residue for later use;
drying the decocted filter residue in a 55 ℃ oven for 15min, placing the dried filter residue in a reaction kettle, adding 7 times of ammonia water (the concentration is 30%) by mass, sealing and soaking for 2h, heating to 115 ℃, reacting for 20h, cooling to room temperature, opening the reaction kettle, filtering, and taking the filter residue for later use;
adjusting the pH of the filter residue after the alkali liquor reaction to 6.0 by using dilute sulfuric acid, adding cellulase, cellobiase and alpha-amylase (the adding proportions are respectively 18IU/g, 7IU/g and 9IU/g), placing the mixture in an environment at 40 ℃ for reaction for 22 hours, and filtering the mixture to obtain an enzymatic hydrolysate;
adjusting pH of the enzymatic hydrolysate to 7.8 with magnesium carbonate, adding 45mg/L vitamin C, sterilizing at 120 deg.C for 24min, inoculating 8% Actinobacillus succinogenes F3-21, fermenting at 35 deg.C, and continuously introducing 100% CO during fermentation 2 And carrying out anaerobic fermentation for 55h, and finally measuring that the concentration of the succinic acid in the fermentation product reaches 78.6 g/L.
Example 6
A method for producing succinic acid by fermentation comprises the following steps:
drying the residue (containing equal mass of calculus bovis, rhizoma corydalis, radix Angelicae sinensis, pericarpium Citri Tangerinae, fructus crataegi, Atractylodis rhizoma and Raphani semen) in oven at 65 deg.C to constant weight, mixing 500g with 5.5kg purified water, adding into ultrasonic instrument, and stirring at 6W/m 2 Performing ultrasonic treatment for 20min at the intensity of (1), taking out, putting into a reaction kettle, heating to 135 ℃, decocting for 25min, cooling to room temperature, filtering, and taking filter residue for later use;
drying the decocted filter residue in a 65 ℃ oven for 20min, placing the dried filter residue in a reaction kettle, adding 6 times of ammonia water (the concentration is 35%) by mass, sealing and soaking for 2h, heating to 110 ℃, reacting for 20h, cooling to room temperature, opening the reaction kettle, filtering, and taking the filter residue for later use;
adjusting the pH of the filter residue after the alkali liquor reaction to 6.1 by using dilute sulfuric acid, adding cellulase, cellobiase and alpha-amylase (the adding proportions are 22IU/g, 10IU/g and 5IU/g respectively), placing the mixture in an environment at 40 ℃ for reaction for 22 hours, and filtering the mixture to obtain an enzymatic hydrolysate;
adjusting pH of the enzymatic hydrolysate to 7.0 with magnesium carbonate, adding 40mg/L vitamin C, sterilizing at 125 deg.C for 25min, inoculating 7% Actinobacillus succinogenes F3-21, fermenting at 38 deg.C, and introducing 100% CO continuously during the fermentation process 2 And carrying out anaerobic fermentation for 58h, and finally measuring that the concentration of the succinic acid in the fermentation product reaches 80.8 g/L.
Examples of the experiments
This example explores the effect of different enzymatic pretreatments on fermentation yield.
This example was set up with 5 experimental groups, which were treated substantially as in example 1, with the following differences: the experiment group 1 does not carry out ultrasonic treatment on the traditional Chinese medicine residues; the experimental group 2 does not heat and boil the traditional Chinese medicine residues at high temperature; the experimental group 3 omits the step of treating the first filter residue by ammonia water, and directly carries out enzymolysis on the filter residue after heating and boiling; in the experimental group 4, the filter residue after heating and boiling is soaked, sealed and heated by ammonium sulfate; experimental group 5 used sodium hydroxide solution instead of ammonia solution.
The results of calculating the yield of succinic acid at 60 th fermentation time by subjecting the enzymatic hydrolysates obtained in 5 experimental groups to the same fermentation treatment are shown in table 1.
TABLE 1
| Experimental group | Treatment of | Succinic acid yield |
| 1 | Without ultrasonic treatment | 54.6g/L |
| 2 | Is not subjected to heating and decocting treatment | 52.3g/L |
| 3 | Without ammonia water heating treatment | 48.9g/L |
| 4 | Replacing ammonia water with ammonium sulfate solution | 53.2g/L |
| 5 | Replacing ammonia water with sodium hydroxide solution | 54.5g/L |
As can be seen from the comparison between table 1 and example 1, the lack of partial enzymolysis pretreatment in experimental groups 1 to 3 has a great influence on the final yield of the fermented succinic acid, and the final yield of succinic acid is reduced compared to example 1 because the acid solution is used to replace the alkali solution in experimental group 4 and the sodium hydroxide solution is used to replace the ammonia solution in experimental group 5.
In summary, the method for producing succinic acid by fermentation according to the embodiment of the present application. The method adopts the traditional Chinese medicine residues as a carbon source for producing succinic acid by fermentation, firstly carries out ultrasonic treatment and heating boiling on the traditional Chinese medicine residues, can destroy the structure of lignocellulose in the traditional Chinese medicine residues under ultrasonic and high temperature, breaks the connection of partial cellulose and hemicellulose, and destroys an inner layer structure containing a large amount of lignin; on the basis, ammonia water is added to fully soak the lignocellulose, the lignocellulose is sealed and heated, the ammonia water can effectively soak the lignocellulose in the environment of high-temperature alkali liquor, chemical bonding between the lignin and the hemicellulose is completely destroyed, the polymerization degree of the lignin and the hemicellulose is reduced, most of the lignin and the hemicellulose surrounding the cellulose are removed, a cellulose crystallization area is fully exposed, the contact area between the cellulose and cellulase molecules is increased, the subsequent enzymolysis part is facilitated to carry out full enzymolysis, the fermentation effect is improved, the conversion rate of succinic acid is improved, and the yield of the final succinic acid is over 56%. This application adopts aqueous ammonia solution as alkali liquor heat treatment's raw materials, can improve the enzymolysis effect on the one hand, and on the other hand aqueous ammonia easily removes, has reduced subsequent sewage treatment process. The method adopts the Chinese medicine residues as the raw materials for producing the succinic acid by fermentation, makes full use of a large amount of cellulose and starch in the raw materials, and plays a role in resource recycling.
The embodiments described above are some, but not all embodiments of the present application. The detailed description of the embodiments of the present application is not intended to limit the scope of the claimed application, but is merely representative of selected embodiments of the application. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.
Claims (10)
1. The method for producing the succinic acid by fermentation is characterized by comprising the following steps of:
adding water into the Chinese medicinal residue, performing ultrasonic treatment and heating and decocting, and performing solid-liquid separation to obtain first residue;
drying the first filter residue, mixing with ammonia water, soaking, sealing, heating, and performing solid-liquid separation to obtain a second filter residue;
carrying out enzymolysis on the second filter residue, and carrying out solid-liquid separation to obtain an enzymolysis liquid;
adjusting the pH of the enzymolysis liquid to be neutral, sterilizing, inoculating zymocyte, and fermenting to produce the succinic acid.
2. The method for producing succinic acid by fermentation according to claim 1, wherein the mass ratio of the herb residue to water is 1: (8-12), the ultrasonic intensity is 5-8W/m 2 And the ultrasonic time is 10-30 min.
3. The method for producing succinic acid by fermentation according to claim 2, wherein the heating and boiling temperature is 130 to 150 ℃ for 20 to 40 min.
4. The method for producing succinic acid by fermentation according to claim 1, wherein the first filter residue is dried at 50-70 ℃ for 8-15 min.
5. The method for producing succinic acid by fermentation according to claim 4, wherein the solid-liquid mass ratio of the dried first residue to the ammonia water is 1: (6-10), wherein the concentration of the ammonia water is 25-35%.
6. The method for producing succinic acid by fermentation according to claim 5, wherein the soaking time is 1-3 hours, the sealing and heating temperature is 100-120 ℃, and the heating time is 10-20 hours.
7. The method for producing succinic acid by fermentation according to claim 1, wherein the step of enzymatic hydrolysis specifically comprises: and adjusting the pH value of the second filter residue to 5.5-6.6, adding cellulase, cellobiase and alpha-amylase, and carrying out enzymolysis for 16-28 h at 35-40 ℃.
8. The method for producing succinic acid by fermentation according to claim 7, wherein the cellulase is added in an amount of 15 to 25IU/g, cellobiase is added in an amount of 5 to 10IU/g, and alpha-amylase is added in an amount of 5 to 10 IU/g.
9. The method for producing succinic acid by fermentation according to claim 1, wherein the pH is adjusted to 6.5-7.8 by using magnesium carbonate, the temperature for sterilization is 120-125 ℃, and the time for sterilization is 20-25 min.
10. The method of claim 9The method for producing succinic acid by fermentation is characterized in that the fermentation bacteria are actinobacillus succinogenes F3-21, the inoculation amount is 4-8%, and 100% CO is introduced in the fermentation process 2 And the fermentation temperature is 35-38 ℃.
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