CN110904169A - Efficient green production process of amino acid - Google Patents
Efficient green production process of amino acid Download PDFInfo
- Publication number
- CN110904169A CN110904169A CN201911317193.2A CN201911317193A CN110904169A CN 110904169 A CN110904169 A CN 110904169A CN 201911317193 A CN201911317193 A CN 201911317193A CN 110904169 A CN110904169 A CN 110904169A
- Authority
- CN
- China
- Prior art keywords
- fermentation
- fermentation medium
- raw materials
- medium
- taking
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- 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
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/04—Alpha- or beta- amino acids
- C12P13/14—Glutamic acid; Glutamine
- C12P13/18—Glutamic acid; Glutamine using biotin or its derivatives
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
Abstract
The invention belongs to the technical field of amino acid production, and discloses an efficient green manufacturing process of amino acid, which comprises the following steps: step 1) preparing an optimized fermentation medium, step 2) fermenting process, and step 3) extracting and separating process. The invention has high fermentation efficiency, improves the separation and extraction process, improves the product recovery rate and purity, reduces energy consumption and sewage discharge, and meets the requirement of green manufacturing.
Description
Technical Field
The invention belongs to the technical field of amino acid production, and particularly relates to an efficient green manufacturing process of amino acid.
Background
Amino acids play an increasingly widespread role in the food industry, medicine, agriculture, animal husbandry, and in the human health, health care, cosmetic industry, etc. Statistically, the variety of amino acids and their derivatives has been developed from about 50 in the 60's of the 20 th century to over 1000 at present. At present, the world has over 600 million tons of amino acids, and the production is expanded and transferred to developing countries. The international amino acid science society promulgates survey reports that the asia-pacific region has become the largest amino acid market worldwide. China is a large country for producing and consuming amino acids, is in the front of the world in terms of total industrial yield and annual output value, and plays an important role in national economic development. However, compared with other industries or foreign amino acid industries, the amino acid industry in China has the problems of few innovative products, unreasonable product structure, backward main production technical indexes, large energy consumption, serious environmental pollution, higher production cost and the like, and particularly, the requirements of international standards cannot be met in the aspects of hardware such as production equipment and the like. Therefore, although China is a large country for producing and consuming amino acids, China is difficult to be called a real strong country of amino acids.
The amino acid production method includes extraction, chemical synthesis, enzyme method and microbial fermentation. The fermentation method has the advantages of low raw material cost, mild reaction conditions, easy realization of large-scale production and the like. At present, in China, except for L-histidine, partial L-leucine and partial L-cysteine are produced by an extraction method, L-serine and partial L-cysteine are produced by an enzyme method, and other amino acid products are produced by a fermentation method. Compared with the foreign amino acid industry, the main production technical indexes of the amino acid fermentation industry in China are relatively lagged behind, and the distance from the truly strong fermentation country is not small.
Although the amino acid fermentation technology in China has made remarkable progress. But the technological innovation capability is still weak, and the technical level needs to be improved. Although the research and development rate is obviously increased in recent years, the method is still relatively insufficient. Technology is an important limiting factor in the development of the amino acid industry behind other industries. China only depends on energy and has low production cost advantage and is difficult to compete with the large country for producing amino acid. Therefore, key technologies must be broken through, autonomous innovation capability must be enhanced, and an effective technical support system and a quality assurance system must be established.
Glutamic acid is the amino acid with the largest yield in amino acids, is mainly prepared by microbial fermentation, and can improve the thallus multiplication rate and the yield of the amino acids by optimizing a fermentation culture medium. In the prior art, a great deal of research is carried out on the optimization of a glutamic acid fermentation medium, for example, in document 1, "the fermentation medium of corynebacterium glutamicum CNl021 is optimized based on a PB test and a response surface analysis method, and in 2014, the fermentation medium composition is optimized by a steepest-grade test and a response surface analysis method to obtain an optimal fermentation medium group of corynebacterium glutamicum, and the acid yield of the fermentation medium is increased by 22.75% compared with that of a fermentation medium of an unoptimized medium. Document 2 "influence of mixed rare earth nitrates on glutamic acid-producing bacteria, amino acid journal" found that addition of a certain amount of rare earth elements to a fermentation medium can increase the yield of glutamic acid. The prior patent technology of the applicant, namely 'a preparation method of a glutamic acid fermentation culture medium', is improved on the basis of a conventional culture medium, and a yeast extract nitrogen source is replaced by adding a mycoprotein extract, so that the cost is saved, the amino acid fermentation yield can be improved, and two purposes are achieved.
The extraction process of the glutamic acid mainly has the following disadvantages: 1. the prior domestic amino acid production still adopts the traditional technical means of plate frame, ion exchange, decoloration, drying and the like, the product has low purity, large acid and alkali dosage and serious environmental pollution, and becomes a key factor for restricting the green development of the biological fermentation industry. 2. The traditional flocculating agent causes the technical problems of high baume degree and viscosity of waste liquid and high difficulty in waste water treatment. 3. In the drying process, the feed liquid is easy to generate coking phenomenon, the quality of the product is influenced, and bad smell is generated.
Disclosure of Invention
The invention discloses a method for producing α -type glutamic acid by fermentation, which improves the fermentation acid production efficiency by optimizing a fermentation process, and on the basis, the applicant continuously improves a separation and extraction process to achieve the technical aims of improving the yield, reducing the energy consumption and reducing the sewage discharge.
The invention is realized by the following technical scheme.
An efficient green process for the manufacture of amino acids comprising the steps of:
step 1) preparing an optimized fermentation medium, step 2) fermenting process, and step 3) extracting and separating process.
Further, the optimized fermentation medium comprises a fermentation medium A and a fermentation medium B which are used separately;
the fermentation medium A is prepared by the following method: taking the following raw materials: glucose, Yeast extract, K2HPO4，MgSO4·7H2O, 2-hydroxyethylamine, CeCl3，MnSO4·H2O，FeSO4·7H2O，VB1Biotin; stirring the raw materials uniformly, adjusting the pH value, and sterilizing to obtain a fermentation medium A;
the fermentation medium B is prepared by the following method:
taking the following raw materials: succinic acid, urea, chitosan; and (3) uniformly stirring all the raw materials, adjusting the pH value, and sterilizing to obtain a fermentation medium B.
Further, the fermentation process in the step 2) specifically comprises the following steps: inoculating the seed solution of Brevibacterium flavum for producing glutamic acid into a 100L fermentation tank filled with 60L fermentation medium A according to the inoculation amount of 8-10% for fermentation culture for 24h, then adding 10L fermentation medium B, continuing the fermentation culture for 24h, and collecting the fermentation liquor; in the whole fermentation culture process, the fermentation temperature is controlled to be 30-36 ℃, the ventilation ratio is 1: 0.7-0.9, the stirring speed is 200-.
Further, the extraction and separation process in the step 3) specifically comprises the following steps: taking fermentation liquor, centrifuging by adopting a high-speed disc centrifuge, and collecting filtrate and mycoprotein precipitation; adding 0.5-2% of flocculating agent into the filtrate, standing for 6-12h, filtering with a plate frame, and collecting liquid; then filtering the mixture by a ceramic membrane, and collecting filtrate; introducing the filtrate into a decolorizing tank with the addition of active carbon of 0.5-1% for 30-120min, filtering, and collecting decolorized solution; carrying out secondary decolorization on the decolorized solution through a decolorizing membrane, and collecting decolorized clear liquid; carrying out chromatographic separation on the decolorized clear liquid through a sequential simulated moving bed to obtain an extracting solution; and (3) carrying out four-effect concentration, centrifugation and fluidized bed spray drying on the extracting solution to obtain the compound.
Preferably, the preparation method of the fermentation medium A comprises the following steps: taking the following raw materials: glucose 50-100g/L, yeast extract 10-30g/L, K2HPO41-5g/L，MgSO4·7H2O20-200 mg/L, 2-hydroxyethylamine 10-50mg/L, CeCl31-20mg/L，MnSO4·H2O 1-10mg/L，FeSO4·7H2O 1-10mg/L，VB15-50mg/L, biotin 1-10 mug/L; stirring the raw materials uniformly, adjusting pH to 6-7, sterilizing at 121 deg.C, and naturally cooling to obtain fermentation culture medium A.
Preferably, the preparation method of the fermentation medium A comprises the following steps:
taking the following raw materials: 80g/L glucose, 20g/L yeast extract, K2HPO42g/L，MgSO4·7H2O50 mg/L, 2-hydroxyethylamine 40mg/L, CeCl310mg/L，MnSO4·H2O 3mg/L，FeSO4·7H2O 3mg/L，VB110mg/L, biotin 7 mu g/L; stirring the raw materials uniformly, adjusting pH to 6.5, sterilizing at 121 deg.C for 15min, and naturally cooling to obtain fermentation culture medium A.
Preferably, the preparation method of the fermentation medium B comprises the following steps:
taking the following raw materials: 1-10g/L of succinic acid, 1-5g/L of urea and 20-100mg/L of chitosan; stirring the raw materials uniformly, adjusting the pH value, and sterilizing to obtain a fermentation medium B;
preferably, the preparation method of the fermentation medium B comprises the following steps:
taking the following raw materials: 5g/L of succinic acid, 2g/L of urea and 80mg/L of chitosan; stirring the raw materials uniformly, adjusting pH to 6.5, sterilizing at 121 deg.C for 15min, and naturally cooling to obtain fermentation medium B.
Preferably, the flocculant is prepared by mixing chitosan and sodium alginate according to the mass ratio of 2: 1.
Compared with the prior art, the invention has the advantages that the following aspects are mainly included but not limited:
the fermentation medium of the invention consists of two parts, wherein the fermentation medium A emphasizes on the improvement of the proliferation of strains, and the fermentation medium B emphasizes on the synthesis and the secretion of glutamic acid;
during early cell proliferation, 2-hydroxyethylamine can promote synthesis of components of the cell wall of phosphatidylethanolamine, so that the proliferation rate of the strain is increased, the later strain proliferation is slow, acid production is mainly performed, and 2-hydroxyethylamine can also be used as a cationic surfactant, so that the cell wall is loosened, the cell permeability is improved, and glutamic acid is promoted to be released to the outside of the cell;
CeCl3the rare earth salt can promote the proliferation of strains, improve the activity of the related synthetase of the glutamic acid and improve the yield of the glutamic acid; however, the excessive concentration can cause the strains to proliferate and die, and the yield of the glutamic acid is correspondingly reduced;
in the middle and later period of fermentation, the proliferation speed of the strain is slowed down, acid production is taken as the main part, amino on chitosan is combined with teichoic acid or lipopolysaccharide with negative charges in the bacterial cell wall, and metal cations are chelated, so that the permeability of the cell wall is changed, and the glutamic acid is promoted to be secreted out of cells.
Succinic acid is added into a fermentation medium, so that the tricarboxylic acid cycle has a certain promotion effect, and the glyoxylate cycle pathway is inhibited, so that the intermediate metabolite flows to the tricarboxylic acid cycle pathway more, and the increase of the glutamic acid yield is promoted.
The method takes the final aims of obtaining the high-purity and high-cost-performance amino acid product, reducing the environmental pollution and improving the greening level, adopts a comprehensive green separation and extraction technology taking a chromatography and a multistage membrane coupling separation and purification technology as the core, greatly reduces the acid-base usage amount and the water consumption in the extraction process, reduces the energy consumption in the extraction process, and reduces the generation amount of high ammonia-nitrogen wastewater in the extraction process; the fine filtration of the amino acid fermentation liquor is realized by a ceramic membrane and decolorizing membrane filtration technology, so that the precision requirement of the simulated moving bed chromatography is met; the novel chitin and sodium alginate flocculant is green and efficient, replaces the traditional polyacrylamide, obviously improves the protein yield, reduces the baume degree and viscosity of waste liquid, and realizes the breakthrough of greening of industrial flocculant.
Drawings
FIG. 1: CeCl3Influence of rare earth salts on the concentration of the bacteria;
FIG. 2: CeCl3Influence of rare earth salts on glutamic acid content;
FIG. 3: influence of 2-hydroxyethylamine on the concentration of the bacteria;
FIG. 4: the effect of 2-hydroxyethylamine on glutamic acid content;
FIG. 5: influence of 2-hydroxyethylamine on the conversion of sugar acids.
Detailed Description
In order to make those skilled in the art better understand the technical solutions in the present application, the technical solutions in the present application will be clearly and completely described below with reference to specific embodiments of the present application, and it is obvious that the described embodiments are only a part of the embodiments of the present application, and not all embodiments. 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 invention.
Example 1
An efficient green process for the manufacture of amino acids comprising the steps of:
1) an optimized glutamic acid fermentation medium which comprises a fermentation medium A and a fermentation medium B; the fermentation medium A was added first and then the fermentation medium B was added at 24h intervals.
The preparation method of the fermentation medium A comprises the following steps: taking the following raw materials: 80g/L glucose, 20g/L yeast extract, K2HPO42g/L，MgSO4·7H2O50 mg/L, 2-hydroxyethylamine 40mg/L, CeCl310mg/L，MnSO4·H2O 3mg/L，FeSO4·7H2O 3mg/L，VB110mg/L, biotin 7 mu g/L; stirring the raw materials uniformly, adjusting pH to 6.5, sterilizing at 121 deg.C for 15min, and naturally cooling to obtain fermentation culture medium A;
the preparation method of the fermentation medium B comprises the following steps: taking the following raw materials: 5g/L of succinic acid, 2g/L of urea and 80mg/L of chitosan; stirring the raw materials uniformly, adjusting pH to 6.5, sterilizing at 121 deg.C for 15min, and naturally cooling to obtain fermentation medium B;
2) the fermentation process comprises the following steps: mixing GDK-9 seed solution (OD) of Brevibacterium flavum600nm13.5) inoculating the mixture into a 100L fermentation tank filled with 60L of fermentation medium A according to the inoculation amount of 8 percent for fermentation culture for 24 hours, then adding 10L of fermentation medium B, continuing the fermentation culture for 24 hours, and collecting fermentation liquor; in the whole fermentation culture process, the fermentation temperature is controlled to be 35 ℃, the ventilation ratio is 1: 0.7, the stirring speed is 300r/min, the dissolved oxygen is maintained at 20 percent, glucose with the fed-batch mass percentage concentration of 20 percent is fed-batch to maintain the residual sugar to be not less than 1.0 percent, and the fed-batch defoaming agent is fed-batch for defoaming;
3) the extraction and separation process comprises the following steps:
taking fermentation liquor, centrifuging for 5min at 4000rpm by adopting a high-speed disc centrifuge, and collecting filtrate and mycoprotein precipitation; adding 1% (mass volume ratio, adding 1g flocculant in 100ml filtrate) of flocculant (chitosan and sodium alginate mixed according to the mass ratio of 2: 1) into the filtrate, mixing, standing for 12h, filtering with a plate frame, and collecting liquid; then filtering the mixture by a ceramic membrane, and collecting filtrate; introducing the filtrate into a decolorizing tank with active carbon addition of 0.5% (mass to volume ratio, 0.5g added into 100ml filtrate) for 60min, filtering, and collecting decolorized solution; carrying out secondary decolorization on the decolorized solution through a decolorizing membrane, and collecting decolorized clear liquid; carrying out chromatographic separation on the decolorized clear liquid through a sequential simulated moving bed to obtain an extracting solution; and (3) carrying out four-effect concentration, centrifugation and fluidized bed spray drying on the extracting solution to obtain the compound.
Compared with the conventional ion-exchange process, the extraction yield of the product is improved by more than 10%, the product purity is more than 99%, the acid and alkali usage amount in the extraction process is reduced by more than 40%, the water consumption is greatly reduced by more than 20%, and the high-ammonia-nitrogen wastewater production amount in the extraction process is reduced by more than 50%; the annual discharge of amino acid wastewater of enterprises is reduced by 25 ten thousand meters3The reuse rate of the waste water reaches more than 70 percent, the energy is saved, the emission is reduced, and the pressure of environmental pollution is reduced.
Example 2
An efficient green process for the manufacture of amino acids comprising the steps of:
1) an optimized glutamic acid fermentation medium which comprises a fermentation medium A and a fermentation medium B; the fermentation medium A was added first and then the fermentation medium B was added at 24h intervals.
The preparation method of the fermentation medium A comprises the following steps: taking the following raw materials: 100g/L glucose, 25g/L yeast extract, K2HPO41g/L，MgSO4·7H2O70 mg/L, 2-hydroxyethylamine 20mg/L, CeCl35mg/L，MnSO4·H2O 2mg/L，FeSO4·7H2O 2mg/L，VB15mg/L, biotin 5 mu g/L; stirring the raw materials uniformly, adjusting pH to 6.5, sterilizing at 121 deg.C for 15min, and naturally cooling to obtain fermentation culture medium A;
the preparation method of the fermentation medium B comprises the following steps: taking the following raw materials: 7g/L of succinic acid, 2g/L of urea and 50mg/L of chitosan; stirring the raw materials uniformly, adjusting pH to 6.5, sterilizing at 121 deg.C for 15min, and naturally cooling to obtain fermentation medium B;
2) the fermentation process comprises the following steps: inoculating Brevibacterium flavum GDK-9 with 8% of inoculum size to obtain seed solution (OD)600nm13.5) inoculating into a 100L fermentation tank filled with 60L fermentation medium A for fermentation culture for 24h, then adding 10L fermentation medium B, continuing fermentation culture for 24h, and collecting fermentation liquor; in the whole fermentation culture process, the fermentation temperature is controlled to be 35 ℃, the ventilation ratio is 1: 0.7, the stirring speed is 300r/min, the dissolved oxygen is maintained at 20 percent, glucose with the fed-batch mass percentage concentration of 20 percent is fed-batch to maintain the residual sugar to be not less than 1.0 percent, and the fed-batch defoaming agent is fed-batch for defoaming;
3) the extraction and separation process comprises the following steps:
taking fermentation liquor, centrifuging for 4min at 5000rpm by adopting a high-speed disc centrifuge, and collecting filtrate and mycoprotein precipitation; adding 1.5% (mass volume ratio, adding 1.5g flocculant in 100ml filtrate) of flocculant (prepared by mixing chitosan and sodium alginate according to the mass ratio of 3: 2) into the filtrate, mixing, standing for 9h, filtering with a plate frame, and collecting liquid; then filtering the mixture by a ceramic membrane, and collecting filtrate; introducing the filtrate into a decolorizing tank with the addition of active carbon of 0.5% (mass volume ratio, adding 0.5g into 100ml filtrate), decolorizing for 90min, filtering and collecting decolorized solution; carrying out secondary decolorization on the decolorized solution through a decolorizing membrane, and collecting decolorized clear liquid; carrying out chromatographic separation on the decolorized clear liquid through a sequential simulated moving bed to obtain an extracting solution; and (3) carrying out four-effect concentration, centrifugation and fluidized bed spray drying on the extracting solution to obtain the compound.
Example 3
Mono, CeCl3Influence of rare earth salts on thallus concentration, glutamic acid content and saccharic acid conversion rate.
The fermentation medium is as follows: 80g/L glucose, 20g/L yeast extract, K2HPO42g/L，MgSO4·7H2O 50mg/L，CeCl30-40mg/L，MnSO4·H2O 3mg/L，FeSO4·7H2O 3mg/L，VB110mg/L, biotin 7 mu g/L;
the fermentation process was the same as in example 1, with 70L of fermentation medium in a 100L fermentor.
Setting up CeCl3Is added at a concentration of 0, 2.5,5,10,20,40mg/L, as shown in FIGS. 1-2, with CeCl3The addition amount is increased, the thallus concentration and the glutamic acid content are both improved, when the addition amount is 10mg/L, the thallus concentration and the glutamic acid content reach peak values, then a descending trend appears, but the saccharic acid conversion rate is not obviously changed in the whole process (not shown in the attached drawing); description of CeCl3The rare earth salt can promote the strain proliferation, improve the activity of the related synthetase of the glutamic acid and improve the yield of the glutamic acid, but the over-high concentration can cause the strain proliferation to be slow and die, and the yield of the glutamic acid is correspondingly reduced.
Second, the determination of CeCl by the above experiment3The effect of 2-hydroxyethylamine on the cell concentration, glutamic acid content and conversion rate of saccharic acid was investigated on the basis of the addition amount of 10 mg/L. Setting the concentration of 2-hydroxyethylamine to be 2.5,5,10,20,40, 80 and 160mg/L, as shown in fig. 3-4, increasing the thallus concentration with the increase of the addition amount of 2-hydroxyethylamine, correspondingly increasing the glutamic acid content and the saccharic acid conversion rate, when the addition amount is 40mg/L, the thallus concentration and the glutamic acid content reach peak values, continuously increasing the concentration of 2-hydroxyethylamine, generating obvious bacteriostasis effect and obviously reducing the density of the strain; the reason is that 2-hydroxyethylamine may promote phosphatidylethanolamine cell wall groupsThe bacterial strain proliferation rate is improved, but the bacterial strain proliferation phenomenon can be caused due to the excessive concentration, the later bacterial strain proliferation is slowed down, the acid production is taken as the main part, and the 2-hydroxyethylamine can also be taken as a cationic surfactant, so that the cell wall is loosened, the cell permeability is improved, the glutamic acid is promoted to be released to the fermentation liquor, and the glutamic acid yield and the sugar acid conversion rate are improved.
Thirdly, the determination of CeCl by the above experiment3The influence of the fermentation medium B on the cell concentration, glutamic acid content and sugar-acid conversion rate was investigated on the basis of 10mg/L and 40mg/L of 2-hydroxyethylamine.
The control group 1 was fermented with fermentation medium A without fermentation medium B, and a 100L fermenter contained 70L fermentation medium A; the fermentation process is referred to example 1.
Control group 2: the fermentation medium B was the same as in example 1 except that succinic acid was not added.
Control group 3: the fermentation medium B was the same as in example 1 except that chitosan was not added.
Control group 4: 5g/L succinic acid was added to the fermentation medium A, and the remainder was the same as in control 1.
The experimental group is example 1.
Specific results are shown in table 1.
TABLE 1
| Group of | Bacterial concentration OD600nm | Glutamic acid output g/L | Conversion rate of sugar and acid% |
| Control group 1 | 53.9 | 137.4 | 59.1 |
| Control group 2 | 54.2 | 141.2 | 61.8 |
| Control group 3 | 54.5 | 145.8 | 62.6 |
| Control group 4 | 53.8 | 137.9 | 59.2 |
| Experimental group | 55.3 | 150.7 | 64.1 |
And (4) conclusion: the control group 1 adopts a single fermentation medium for fermentation, the yield of glutamic acid and the conversion rate of saccharic acid are obviously lower than those of the experimental group, and the concentration difference of thalli is not large; the control group 4 is added with succinic acid on the basis of the control group 1, so that the concentration of thalli is not influenced, and the yield of glutamic acid and the conversion rate of saccharic acid are not obviously different, probably because the thalli proliferation is taken as the main part in the early stage of fermentation, the acid production is less, and the succinic acid has no obvious stimulation effect on the thalli proliferation; the experimental group and the control group 3 adopt succinic acid added in the middle of fermentation, at the moment, the proliferation of thalli is slow, acid production is mainly performed, succinic acid has positive promotion effect on tricarboxylic acid cycle and has inhibition effect on glyoxylic acid cycle, so that the yield of glutamic acid is increased; gradient tests show that the addition amount of succinic acid is too large (larger than 10 g/L), the yield of glutamic acid cannot be further improved, comprehensive cost is considered, and the addition amount of succinic acid lower than 10g/L is more suitable. The chitosan is added in the middle and later fermentation stages of the control group 2 and the experimental group, so that the permeability of cell walls can be changed, and the secretion of glutamic acid to the outside of cells is promoted, thereby improving the yield of the glutamic acid and the conversion rate of saccharic acid; however, the excessive addition amount (more than 100 mg/L) of the chitosan can cause the occurrence of bacteriostasis, thereby causing the death of the strain.
Although the invention has been described in detail with respect to the general description and the specific embodiments, it will be apparent to those skilled in the art that modifications and improvements can be made to the invention or the method can be practiced without the specific embodiments. Accordingly, it is intended that all such modifications, improvements and extensions that do not depart from the spirit of the invention, be considered within the scope of the invention as claimed.
Claims (10)
1. An efficient green process for the manufacture of amino acids comprising the steps of:
step 1) preparing an optimized fermentation medium, step 2) fermenting process, and step 3) extracting and separating process.
2. The manufacturing process of claim 1, wherein the optimized fermentation medium comprises fermentation medium a and fermentation medium B used separately;
the fermentation medium A is prepared by the following method: taking the following raw materials: glucose, Yeast extract, K2HPO4，MgSO4·7H2O, 2-hydroxyethylamine, CeCl3，MnSO4·H2O，FeSO4·7H2O，VB1Biotin; stirring the raw materials uniformly, adjusting the pH value, and sterilizing to obtain a fermentation medium A;
the fermentation medium B is prepared by the following method:
taking succinic acid, urea and chitosan; and (3) uniformly stirring all the raw materials, adjusting the pH value, and sterilizing to obtain a fermentation medium B.
3. The manufacturing process according to claims 1-2, wherein the step 2) fermentation process specifically comprises: inoculating the seed solution of Brevibacterium flavum for producing glutamic acid into a 100L fermentation tank filled with 60L fermentation medium A according to the inoculation amount of 8-10% for fermentation culture for 24h, then adding 10L fermentation medium B, continuing the fermentation culture for 24h, and collecting the fermentation liquor; in the whole fermentation culture process, the fermentation temperature is controlled to be 30-36 ℃, the ventilation ratio is 1: 0.7-0.9, the stirring speed is 200-.
4. The manufacturing process according to claims 1 to 3, wherein the step 3) of extraction and separation comprises in particular: taking fermentation liquor, centrifuging by adopting a high-speed disc centrifuge, and collecting filtrate and mycoprotein precipitation; adding 0.5-2% of flocculating agent into the filtrate, standing for 6-12h, filtering with a plate frame, and collecting liquid; then filtering the mixture by a ceramic membrane, and collecting filtrate; introducing the filtrate into a decolorizing tank with the addition of active carbon of 0.5-1% for 30-120min, filtering, and collecting decolorized solution; carrying out secondary decolorization on the decolorized solution through a decolorizing membrane, and collecting decolorized clear liquid; carrying out chromatographic separation on the decolorized clear liquid through a sequential simulated moving bed to obtain an extracting solution; and (3) carrying out four-effect concentration, centrifugation and fluidized bed spray drying on the extracting solution to obtain the compound.
5. The process according to claims 2-3, wherein fermentation medium A is prepared by: taking the following raw materials: glucose 50-100g/L, yeast extract 10-30g/L, K2HPO41-5g/L，MgSO4·7H2O20-200 mg/L, 2-hydroxyethylamine 10-50mg/L, CeCl31-20mg/L，MnSO4·H2O 1-10mg/L，FeSO4·7H2O 1-10mg/L，VB15-50mg/L, biotin 1-10 mug/L; stirring the raw materials uniformly, adjusting pH to 6-7, sterilizing at 121 deg.C, and naturally cooling to obtain fermentation culture medium A.
6. The process of claim 5, wherein the fermentation medium A is prepared by:
taking the following raw materials: 80g/L glucose, 20g/L yeast extract, K2HPO42g/L，MgSO4·7H2O50 mg/L, 2-hydroxyethylamine 40mg/L, CeCl310mg/L，MnSO4·H2O 3mg/L，FeSO4·7H2O 3mg/L，VB110mg/L, biotin 7 mu g/L; stirring the raw materials uniformly, adjusting pH to 6.5, sterilizing at 121 deg.C for 15min, and naturally cooling to obtain fermentation culture medium A.
7. The process according to claims 2-3, wherein fermentation medium B is prepared by:
taking the following raw materials: 1-10g/L of succinic acid, 1-5g/L of urea and 20-100mg/L of chitosan; and (3) uniformly stirring all the raw materials, adjusting the pH value, and sterilizing to obtain a fermentation medium B.
8. The process of claim 7, wherein fermentation medium B is prepared by:
taking the following raw materials: 5g/L of succinic acid, 2g/L of urea and 80mg/L of chitosan; stirring the raw materials uniformly, adjusting pH to 6.5, sterilizing at 121 deg.C for 15min, and naturally cooling to obtain fermentation medium B.
9. The manufacturing process according to claim 4, wherein the flocculant is prepared by mixing chitosan and sodium alginate according to a mass ratio of 2: 1.
10. Amino acid product obtainable by the manufacturing process according to claims 1-9.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911317193.2A CN110904169B (en) | 2019-12-19 | 2019-12-19 | Efficient green production process of amino acid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911317193.2A CN110904169B (en) | 2019-12-19 | 2019-12-19 | Efficient green production process of amino acid |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110904169A true CN110904169A (en) | 2020-03-24 |
| CN110904169B CN110904169B (en) | 2022-07-15 |
Family
ID=69826547
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911317193.2A Active CN110904169B (en) | 2019-12-19 | 2019-12-19 | Efficient green production process of amino acid |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110904169B (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1903741A (en) * | 2006-08-02 | 2007-01-31 | 袁毅桦 | Preparation method of chitin-alginic acid copolymer flocculant |
| CN101417958A (en) * | 2008-11-26 | 2009-04-29 | 江南大学 | Method for extracting high quality, low material consumption and low waste water glutamic acid |
| CN105063159A (en) * | 2015-09-19 | 2015-11-18 | 内蒙古阜丰生物科技有限公司 | Novel process for extracting glutamic acid through concentration-continuous isoelectric treatment |
| EP2949660A1 (en) * | 2004-12-28 | 2015-12-02 | Ajinomoto Co., Inc. | L-glutamic acid-producing microorganism and a method for producing l-glutamic acid |
| CN109504719A (en) * | 2018-12-19 | 2019-03-22 | 呼伦贝尔东北阜丰生物科技有限公司 | A method of improving glutamic acid acid production rate and recovery rate |
| CN109678235A (en) * | 2019-01-08 | 2019-04-26 | 广东海洋大学 | A kind of natural zwitterion compound sewage flocculant for treating and its application method |
-
2019
- 2019-12-19 CN CN201911317193.2A patent/CN110904169B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2949660A1 (en) * | 2004-12-28 | 2015-12-02 | Ajinomoto Co., Inc. | L-glutamic acid-producing microorganism and a method for producing l-glutamic acid |
| CN1903741A (en) * | 2006-08-02 | 2007-01-31 | 袁毅桦 | Preparation method of chitin-alginic acid copolymer flocculant |
| CN101417958A (en) * | 2008-11-26 | 2009-04-29 | 江南大学 | Method for extracting high quality, low material consumption and low waste water glutamic acid |
| CN105063159A (en) * | 2015-09-19 | 2015-11-18 | 内蒙古阜丰生物科技有限公司 | Novel process for extracting glutamic acid through concentration-continuous isoelectric treatment |
| CN109504719A (en) * | 2018-12-19 | 2019-03-22 | 呼伦贝尔东北阜丰生物科技有限公司 | A method of improving glutamic acid acid production rate and recovery rate |
| CN109678235A (en) * | 2019-01-08 | 2019-04-26 | 广东海洋大学 | A kind of natural zwitterion compound sewage flocculant for treating and its application method |
Non-Patent Citations (3)
| Title |
|---|
| 宋香: "稀土元素对谷氨酸棒杆菌生长及酶活性的影响", 《无锡轻工大学学报》 * |
| 户红通: "谷氨酸清洁发酵工艺研究", 《中国酿造》 * |
| 韩文清，程立坤，孙霞: "谷氨酸的发酵生产实训", 《发酵生产原理及实训技术》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110904169B (en) | 2022-07-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109486876B (en) | Method for fermenting, extracting and purifying threonine | |
| CN107058416A (en) | A kind of zymotechnique of refined glutamic acid | |
| CN101555503A (en) | Method for separating and extracting L-arabinose from waste wood sugar mother liquid from wood sugar production | |
| CN108285911B (en) | Process for extracting L-isoleucine by fermentation | |
| CN104059169B (en) | A kind of hyaluronic purifying technique | |
| CN110904167A (en) | Optimization method of L-threonine fermentation process | |
| CN111004822A (en) | Production process of high-purity threonine | |
| WO2020134688A1 (en) | Method for preparing high-purity hericium erinaceus polysaccharide by fermenting hericium erinaceus, and fermentation medium thereof | |
| CN113321580B (en) | Method for producing malic acid | |
| CN108285914B (en) | Fermentation process of L-tryptophan | |
| CN104232692A (en) | Method for pretreating molasses and application of molasses obtained through method | |
| CN109628518A (en) | A method of production and extraction L-Glutamine | |
| CN113773963A (en) | High-density high-yield porphyridium culture method | |
| CN101823953B (en) | Method for separating lactic acid from fermentation liquor by flocculation and carrying out flocculate comprehensive utilization | |
| CN111139273B (en) | Method for preparing, separating and extracting L-tryptophan | |
| CN103693779B (en) | A kind for the treatment of process of lysine fermentation liquor Waste water concentrating liquid and the method for fermentative production Methionin | |
| CN111057727B (en) | Method for producing, separating and extracting L-glutamine | |
| CN105219804A (en) | A kind of molasses fermented method preparing microbial flocculant | |
| CN110904169B (en) | Efficient green production process of amino acid | |
| CN110885866B (en) | Novel glutamic acid fermentation and monosodium glutamate production method | |
| CN1765897A (en) | Lactochrome fermentation liquor extraction process | |
| CN114958631A (en) | Method for producing single-cell protein by using heavy-phase lactic acid | |
| CN112501077B (en) | Method for culturing clostridium butyricum | |
| CN110592154B (en) | Process for producing and extracting tryptophan | |
| CN103992964A (en) | High pH value tolerant bacterial strain and novel fermentation method for producing lysine |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20220524 Address after: 162650 Kaichuang street, Lingdong Industrial Development Zone (Zhalantun), Hulunbuir City, Inner Mongolia Autonomous Region Applicant after: HULUNBEIER NORTHEAST FUFENG BIOTECHNOLOGIES Co.,Ltd. Address before: 021000 Kaichuang street, Lingdong Industrial Development Zone (Zalantun), Hulunbuir, Inner Mongolia Autonomous Region Hulunbuir northeast Fufeng Biotechnology Co., Ltd Applicant before: Zhao Lankun |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |