CN111363708A - Method for improving butyric acid fermentation level - Google Patents
Method for improving butyric acid fermentation level Download PDFInfo
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- CN111363708A CN111363708A CN202010338530.2A CN202010338530A CN111363708A CN 111363708 A CN111363708 A CN 111363708A CN 202010338530 A CN202010338530 A CN 202010338530A CN 111363708 A CN111363708 A CN 111363708A
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- 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
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- 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
- C12P7/52—Propionic acid; Butyric acids
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Abstract
The invention provides a method for improving the butyric acid fermentation level, and relates to the technical field of butyric acid fermentation. According to the invention, a proper amount of osmotic pressure protective agent is added into a butyric acid fermentation system, so that the osmotic pressure of the thallus cells in high yield and high metal ion concentration is protected, the thallus activity is maintained, the concentration and yield of butyric acid are improved, the energy consumption of fermentation unit is reduced, and the production efficiency is improved. The results show that mannitol is used as a protective agent, the fermentation is improved when the concentration is 3g/L, the effect is not obvious when trehalose is used, and the fermentation efficiency is slightly improved when betaine is used at the concentration of 5 g/L.
Description
Technical Field
The invention belongs to the technical field of butyric acid fermentation, and particularly relates to a method for improving the butyric acid fermentation level.
Background
The butyric acid producing bacteria are anaerobic culture, and the bacteria cannot grow or even die in the presence of oxygen in a fermentation environment; in addition, the cell body is strictly required to have an optimum pH for growth, and if the pH deviates from the optimum pH, the target product may not be obtained. The common butyric acid bacteria fermentation is three-stage fermentation, and the fermentation process at each stage comprises the following steps: the first-stage seed culture process comprises the following steps: the temperature is 37 ℃, the anaerobic reaction is carried out, the rotating speed is 200rpm, the pH is natural, and the culture time is 20-24 h; secondary seed culture: the temperature is 37 ℃, the rotating speed is 200rpm, the culture period is 10-24 h, orp is controlled to be-300 to-500, and the pH is controlled to be 5.90; fermentation culture: the temperature is 37 ℃, the rotating speed is 150-200 rpm, the culture period is 40-50 h, orp is controlled to be-400 to-500, and the pH is controlled to be 5.90; and (4) detecting the content of butyric acid after fermentation is finished, and adding a proper amount of sodium salt or calcium salt according to the concentration of the butyric acid to obtain the butyrate.
The existing fermentation level still has the problems of low fermentation efficiency and high cost, the industrialization level is not high, the product concentration is low, the number of byproducts is large, the product concentration is slowly increased after 40 hours of fermentation, and a substrate culture medium is not completely exhausted.
Disclosure of Invention
In view of the above, the present invention aims to provide a method for improving butyric acid fermentation level, which comprises adding a proper amount of osmotic pressure protective agent into a butyric acid fermentation system, protecting the osmotic pressure of bacterial cells in high yield and high metal ion concentration, maintaining the activity of bacterial cells, improving the concentration and yield of butyric acid, reducing the energy consumption of fermentation unit, and improving the production efficiency.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a method for improving butyric acid fermentation level, wherein an osmotic pressure protective agent is added into a fermentation medium in a butyric acid fermentation process, and the addition amount of the osmotic pressure protective agent is 1-5 g/L.
Preferably, the species of the osmoprotectant includes mannitol, trehalose, or betaine.
Preferably, when mannitol is the osmoprotectant, the addition amount of the osmoprotectant to the fermentation medium is 3 g/L.
Preferably, when betaine is the osmoprotectant, the addition amount of the osmoprotectant to the fermentation medium is 5 g/L.
The invention provides a method for improving butyric acid fermentation level, which is characterized in that a proper amount of osmotic pressure protective agent is added into a butyric acid fermentation system to protect the osmotic pressure of thallus cells in high yield and high metal ion concentration, maintain thallus activity, realize the improvement of butyric acid concentration and yield, reduce the energy consumption of fermentation unit and improve the production efficiency. The osmotic pressure protective agent is a low molecular weight organic compound, is mainly used for reaction to osmotic pressure, can eliminate ROS in a body besides regulating the osmotic pressure, and helps metabolism and detoxification. The osmotic pressure protective agent of the invention is generally present in cytoplasm, inhibits the osmotic imbalance of cells by reducing the osmotic potential of the cells, maintains the turgor pressure of the cells, protects or replaces a water membrane on the surface of the protein under the environment with low water potential and high ionic pressure, and stabilizes the structure of the protein and the membrane. In the embodiment of the invention, mannitol is used as a protective agent, and the concentration of mannitol is 3g/L, so that the mannitol has a promoting effect on fermentation; the fermentation efficiency is slightly improved when the betaine concentration is 5 g/L.
Detailed Description
The invention provides a method for improving butyric acid fermentation level, wherein an osmotic pressure protective agent is added into a fermentation medium in a butyric acid fermentation process, and the addition amount of the osmotic pressure protective agent is 1-5 g/L.
The method for butyric acid fermentation is not particularly limited, and preferably three-stage fermentation, including test tube culture, triangular flask submerged culture, seeding tank expanded culture (two-stage expansion) and fermentation. The culture medium used in the butyric acid fermentation process is not particularly limited, and the conventional butyric acid fermentation culture medium in the field can be used.
The method of the invention only improves the fermentation reaction in the fermentation tank, namely, after the secondary seed solution is inoculated into the fermentation tank, an osmotic pressure protective agent is added into the fermentation tank, and the osmotic pressure protective agent preferably comprises mannitol, trehalose or betaine. In the invention, when mannitol is the osmotic pressure protective agent, the addition amount of the osmotic pressure protective agent to the fermentation medium is preferably 3 g/L; when betaine is the osmotic pressure protective agent, the addition amount of the osmotic pressure protective agent to the fermentation medium is preferably 5 g/L.
In the invention, in a butyric acid fermentation system, a high-concentration product and a high-concentration metal ion are generated along with the continuous fermentation, so that the osmotic pressure is changed, and the osmotic pressure protective agent is added into the butyric acid fermentation system, so that the osmotic pressure of a thallus cell at the time of high product and high metal ion concentration can be protected, the thallus activity is maintained, the concentration and the yield of butyric acid are improved, the energy consumption of a fermentation unit is reduced, and the production efficiency is improved; the osmotic pressure protective agent is a low molecular weight organic compound, is mainly used for the reaction to osmotic pressure, can eliminate ROS in the body besides the regulation to the osmotic pressure, and helps metabolism and detoxification; the osmotic pressure protective agent can be present in cytoplasm, inhibit cell osmotic imbalance by reducing cell osmotic potential, and maintain cell oncotic pressure, protect or replace water membrane on protein surface under low water potential and high ion pressure environment, and stabilize protein and membrane structure.
The method for increasing the fermentation level of butyric acid provided by the present invention is described in detail with reference to the following examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Carrying out test tube culture, triangular flask submerged culture and seeding tank expanded culture on the butyric acid bacteria: inoculating clostridium butyricum in a test tube into 500mL (200 mL) of seed culture medium, and culturing for 10-20 h under an anaerobic condition at the temperature of 37 ℃; putting the butyric acid bacteria cultured in the triangular flask into a 15L seeding tank according to the inoculation amount of 1% for amplification culture; and culturing the secondary seeds for 10-20 hours, transferring the seeds and fermenting.
Shake flask seed culture medium (mass percent): 1% of yeast extract powder, 0.5% of peptone, 0.1% of soluble starch, 0.5% of glucose, 0.01% of manganese chloride, 0.05% of sodium chloride, 0.3% of sodium acetate and the balance of water.
Primary and secondary seed culture media (mass percent): 1% of yeast extract powder, 0.5% of peptone, 0.5% of soluble starch, 1% of glucose and 0.05% of sodium chloride, and the balance of water.
Fermentation medium (mass percent): 2% of glucose, 0.5% of starch, 0.5% of corn starch, 0.1% of ammonium sulfate, 0.5% of peptone, 0.2% of yeast powder, 0.2% of monopotassium phosphate, 0.2% of calcium carbonate, 0.07% of magnesium sulfate, 0.02% of manganese sulfate, 0.1% of defoaming agent and the balance of water.
Under the condition that the original formula of the fermentation basal medium is not changed, the initial osmotic pressure of mannitol fermentation liquor added with an osmotic pressure protective agent is 230mosm/Kg, and different concentrations are compared, and the results are shown in table 1.
TABLE 1 Effect of mannitol on fermentation
Example 2
The conditions were the same as in example 1 except that the osmoprotectant was different, and trehalose was added as the osmoprotectant in this example, and the results are shown in Table 2.
TABLE 2 Effect of trehalose on fermentation
Example 3
The conditions were the same as in example 1 except that the osmoprotectant was different, and betaine was used as the osmoprotectant added in this example, and the results are shown in Table 3.
TABLE 3 influence of betaine on fermentation
In conclusion, the osmotic pressure protective agent provided by the invention has a certain promotion effect on fermentation.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (4)
1. The method for improving the butyric acid fermentation level is characterized in that an osmotic pressure protective agent is added into a fermentation medium in the butyric acid fermentation process, and the addition amount of the osmotic pressure protective agent is 1-5 g/L.
2. The method of claim 1, wherein the species of osmoprotectant comprises mannitol, trehalose, or betaine.
3. The method according to claim 2, wherein when mannitol is the osmoprotectant, the osmoprotectant is added to the fermentation medium in an amount of 3 g/L.
4. The method as claimed in claim 2, wherein when betaine is the osmoprotectant, the addition amount of the osmoprotectant to the fermentation medium is 5 g/L.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114540434A (en) * | 2022-04-27 | 2022-05-27 | 驻马店华中正大有限公司 | Method for improving yield of butyric acid fermentation product |
| CN114540435A (en) * | 2022-04-27 | 2022-05-27 | 驻马店华中正大有限公司 | Method for improving yield of butyric acid by using metabolic enzyme |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101955978A (en) * | 2010-08-31 | 2011-01-26 | 南京工业大学 | Method for enhancing concentration and production intensity of succinic acid by adding osmoprotectant |
| KR20140023721A (en) * | 2012-08-17 | 2014-02-27 | 삼성전자주식회사 | Method of produciton for 1,3-propandiol by adding osmoprotective substance and medium for produciton of 1,3-propandiol |
| CN107201383A (en) * | 2016-03-18 | 2017-09-26 | 中国石化扬子石油化工有限公司 | It is a kind of to improve the D-ALPHA-Hydroxypropionic acid production method that D-ALPHA-Hydroxypropionic acid produces intensity |
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2020
- 2020-04-26 CN CN202010338530.2A patent/CN111363708B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101955978A (en) * | 2010-08-31 | 2011-01-26 | 南京工业大学 | Method for enhancing concentration and production intensity of succinic acid by adding osmoprotectant |
| KR20140023721A (en) * | 2012-08-17 | 2014-02-27 | 삼성전자주식회사 | Method of produciton for 1,3-propandiol by adding osmoprotective substance and medium for produciton of 1,3-propandiol |
| CN107201383A (en) * | 2016-03-18 | 2017-09-26 | 中国石化扬子石油化工有限公司 | It is a kind of to improve the D-ALPHA-Hydroxypropionic acid production method that D-ALPHA-Hydroxypropionic acid produces intensity |
Non-Patent Citations (3)
| Title |
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| DARIA SZYMANOWSKA-POWALOWSKA: ""The effect of high concentrations of glycerol on the growth, metabolism and adaptation capacity of Clostridium butyricum DSP1"", 《ELECTRONIC JOURNAL OF BIOTECHNOLOGY》 * |
| 吴金鹏 等: "《食品微生物学》", 31 October 1998, 农业出版社 * |
| 王友玲 等: ""丁酸梭菌与酪丁酸梭菌发酵特性比较"", 《食品与机械》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114540434A (en) * | 2022-04-27 | 2022-05-27 | 驻马店华中正大有限公司 | Method for improving yield of butyric acid fermentation product |
| CN114540435A (en) * | 2022-04-27 | 2022-05-27 | 驻马店华中正大有限公司 | Method for improving yield of butyric acid by using metabolic enzyme |
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