CN111019862A - Symbiotic bacteria culture medium and symbiotic bacteria culture method - Google Patents
Symbiotic bacteria culture medium and symbiotic bacteria culture method Download PDFInfo
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- CN111019862A CN111019862A CN201911357903.4A CN201911357903A CN111019862A CN 111019862 A CN111019862 A CN 111019862A CN 201911357903 A CN201911357903 A CN 201911357903A CN 111019862 A CN111019862 A CN 111019862A
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Abstract
The embodiment of the invention discloses a symbiotic bacteria culture medium, which comprises the following raw materials in percentage by mass: 2 to 10 percent of molasses, 0.3 to 2 percent of yeast extract, 0.1 to 1 percent of ammonium sulfate, 0.1 to 1 percent of dipotassium hydrogen phosphate, 0.2 to 1 percent of sodium chloride and the balance of water. According to the symbiotic bacteria culture medium and the symbiotic bacteria culture method, lactic acid bacteria, saccharomycetes, bacillus subtilis and photosynthetic bacteria have the advantages of good symbiosis, good stability and high activity in the growth process of the same reaction container.
Description
Technical Field
The embodiment of the invention relates to the technical field of microbial culture, and particularly relates to a symbiotic bacteria culture medium and a symbiotic bacteria culture method.
Background
The mixed culture of various microorganisms has wide application prospect, the mixed system maintains a stable state by original cooperation, mutual assistance and biased cooperation, and antagonism and mutual assistance existing in microbial flora are the traditional Chinese medicine self-stable state of the flora. There is a lack of effective methods and theoretical guidance for the screening and combination of strains with synergistic relationships. The ecological relationship of the complex microbial inoculum is unclear, and the bacteria grow unevenly during mixed culture.
At present, various bacteria are separately cultured and then compounded together in the process for producing the composite microbial preparation, and the composite microbial preparation produced by the method has the problems of poor symbiosis, poor stability, short storage time, low activity and the like. Further improvements are needed.
Disclosure of Invention
Therefore, the embodiment of the invention provides a symbiotic bacteria culture medium and a symbiotic bacteria culture method, which aim to solve the problems of poor symbiosis and poor stability of symbiotic bacteria in the prior art.
In order to achieve the above object, the embodiments of the present invention provide the following technical solutions:
a symbiotic bacteria culture medium comprises the following raw materials in percentage by mass: 2 to 10 percent of molasses, 0.3 to 2 percent of yeast extract, 0.1 to 1 percent of ammonium sulfate, 0.1 to 1 percent of dipotassium hydrogen phosphate, 0.2 to 1 percent of sodium chloride and the balance of water.
Preferably, the symbiotic bacteria culture medium comprises the following raw materials in percentage by mass: 2.2 to 9.8 percent of molasses, 0.5 to 1.8 percent of yeast extract, 0.2 to 0.8 percent of ammonium sulfate, 0.2 to 0.8 percent of dipotassium hydrogen phosphate, 0.4 to 0.8 percent of sodium chloride and the balance of water.
Preferably, the symbiotic bacteria culture medium comprises the following raw materials in percentage by mass: 6% of molasses, 1.2% of yeast extract, 0.5% of ammonium sulfate, 0.4% of dipotassium phosphate, 0.6% of sodium chloride and the balance of water.
Preferably, the pH value of the symbiotic bacteria culture medium is 7.
The embodiment of the invention also provides a symbiotic bacteria culture method, which utilizes the symbiotic bacteria culture medium to culture symbiotic bacteria.
Preferably, the symbiotic bacteria include lactic acid bacteria, bacillus subtilis, yeast and photosynthetic bacteria.
Preferably, the method comprises the steps of:
sterilizing the symbiotic bacteria culture medium at 121 ℃ for 10-20 min;
respectively adding the lactic acid bacteria, the bacillus subtilis, the saccharomycetes and the photosynthetic bacteria into the symbiotic bacteria culture medium;
keeping the temperature of the symbiotic bacteria culture medium at 28-32 ℃, firstly carrying out aerobic fermentation culture for 5-8h, then carrying out anaerobic fermentation culture for 5-8h, repeating the steps, and carrying out culture for 48-80h to obtain a symbiotic culture product.
Preferably, the aerobic fermentation culture conditions are as follows: in the symbiotic bacteria culture medium, sterile air is intermittently introduced according to the aeration ratio of 0.3-0.5, and the stirring speed is 100-150 rpm.
Preferably, the anaerobic fermentation culture conditions are as follows: the symbiotic culture medium was stopped from stirring and the aeration of air was stopped.
The embodiment of the invention has the following advantages:
in the symbiotic bacteria culture medium provided by the embodiment of the invention, the yeast, the bacillus subtilis and the photosynthetic bacteria can quickly grow and reproduce under the aerobic culture condition; under the anaerobic condition, lactic acid bacteria can propagate in a large amount, and photosynthetic bacteria and saccharomycetes can also grow and propagate in a small amount. Wherein, the saccharomycetes utilizes amino acid, sugar and other organic matters to generate fermentation power and generate active substances capable of promoting cell division, thereby creating favorable conditions for the growth and metabolism of photosynthetic bacteria, bacillus subtilis and lactic acid bacteria; lactic acid bacteria can take in substances such as saccharides and amino acids generated by photosynthetic bacteria and yeasts for growth and metabolism. Therefore, the lactobacillus, the saccharomycetes, the bacillus subtilis and the photosynthetic bacteria have good symbiosis, good stability, high activity and the like in the growth process of the same reaction vessel.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. It should be apparent that the drawings in the following description are merely exemplary, and that other embodiments can be derived from the drawings provided by those of ordinary skill in the art without inventive effort.
FIG. 1 is a graph showing the growth of symbiotic bacteria cultured by the symbiotic bacteria culturing method according to the embodiment of the present invention;
Detailed Description
The present invention is described in terms of particular embodiments, other advantages and features of the invention will become apparent to those skilled in the art from the following disclosure, and it is to be understood that the described embodiments are merely exemplary of the invention and that it is not intended to limit the invention to the particular embodiments disclosed. 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.
In the embodiment of the invention, the adopted lactic acid bacteria, saccharomycetes, bacillus subtilis and photosynthetic bacteria are all common commercial strains sold in the market.
Example 1
The embodiment of the invention provides a symbiotic bacteria culture medium, which comprises the following raw materials in percentage by mass: 2% of molasses, 0.3% of yeast extract, 0.1% of ammonium sulfate, 0.1% of dipotassium phosphate, 0.2% of sodium chloride and the balance of water, wherein the pH value of the symbiotic bacteria culture medium is 7.
The embodiment of the invention provides a symbiotic bacteria culture method, which utilizes the symbiotic bacteria culture medium to culture symbiotic bacteria. The symbiotic bacteria adopted in the embodiment of the invention comprise lactic acid bacteria, bacillus subtilis, saccharomycetes and photosynthetic bacteria. The specific culture method of the symbiotic bacteria comprises the following steps: sterilizing the symbiotic bacteria culture medium at 121 ℃ for 10min, and then cooling to normal temperature; then, respectively adding lactic acid bacteria, bacillus subtilis, saccharomycetes and photosynthetic bacteria into a fermentation tank containing a symbiotic bacteria culture medium; keeping the temperature of a fermentation tank containing a symbiotic bacteria culture medium at 28 ℃, and performing aerobic fermentation culture for 5 hours, wherein the aerobic fermentation culture conditions are as follows: introducing sterile air intermittently in the symbiotic bacteria culture medium according to the aeration ratio of 0.3-0, and stirring at the rotation speed of 100 rpm. And then carrying out anaerobic fermentation culture for 5h, wherein the anaerobic fermentation culture conditions are as follows: the symbiotic culture medium was stopped from stirring and the aeration of air was stopped. Repeating the steps for 50h to obtain symbiotic culture product.
Example 2
The embodiment of the invention provides a symbiotic bacteria culture medium, which comprises the following raw materials in percentage by mass: 6% of molasses, 1.2% of yeast extract, 0.5% of ammonium sulfate, 0.4% of dipotassium phosphate, 0.6% of sodium chloride and the balance of water, wherein the pH value of the symbiotic bacteria culture medium is 7.
The embodiment of the invention provides a symbiotic bacteria culture method, which utilizes the symbiotic bacteria culture medium to culture symbiotic bacteria. The symbiotic bacteria adopted in the embodiment of the invention comprise lactic acid bacteria, bacillus subtilis, saccharomycetes and photosynthetic bacteria. The specific culture method of the symbiotic bacteria comprises the following steps: sterilizing a symbiotic bacteria culture medium at 121 ℃ for 15min, and then cooling to normal temperature; then, respectively adding lactic acid bacteria, bacillus subtilis, saccharomycetes and photosynthetic bacteria into a fermentation tank containing a symbiotic bacteria culture medium; keeping the temperature of a fermentation tank containing a symbiotic bacteria culture medium at 30 ℃, and performing aerobic fermentation culture for 6 hours, wherein the aerobic fermentation culture conditions are as follows: sterile air is intermittently introduced into the symbiotic bacteria culture medium according to the aeration ratio of 0.4, and the stirring speed is 125 revolutions per minute. And then carrying out anaerobic fermentation culture for 6h, wherein the anaerobic fermentation culture conditions are as follows: the symbiotic culture medium was stopped from stirring and the aeration of air was stopped. Repeating the steps for 60 hours to obtain a symbiotic culture product.
Example 3
The embodiment of the invention provides a symbiotic bacteria culture medium, which comprises the following raw materials in percentage by mass: 10% of molasses, 2% of yeast extract, 1% of ammonium sulfate, 1% of dipotassium phosphate, 1% of sodium chloride and the balance of water, wherein the pH value of the symbiotic bacteria culture medium is 6.5.
The embodiment of the invention provides a symbiotic bacteria culture method, which utilizes the symbiotic bacteria culture medium to culture symbiotic bacteria. The symbiotic bacteria adopted in the embodiment of the invention comprise lactic acid bacteria, bacillus subtilis, saccharomycetes and photosynthetic bacteria. The specific culture method of the symbiotic bacteria comprises the following steps: sterilizing a symbiotic bacteria culture medium at 121 ℃ for 20min, and then cooling to normal temperature; then, respectively adding lactic acid bacteria, bacillus subtilis, saccharomycetes and photosynthetic bacteria into a fermentation tank containing a symbiotic bacteria culture medium; keeping the temperature of a fermentation tank containing the symbiotic bacteria culture medium at 32 ℃, and firstly carrying out aerobic fermentation culture for 8 hours, wherein the aerobic fermentation culture conditions are as follows: sterile air is intermittently introduced into the symbiotic bacteria culture medium according to the aeration ratio of 0.5, and the stirring speed is 150 rpm. And then carrying out anaerobic fermentation culture for 8h, wherein the anaerobic fermentation culture conditions are as follows: the symbiotic culture medium was stopped from stirring and the aeration of air was stopped. Repeating the steps for 80h to obtain symbiotic culture product.
Test example 1
As shown in fig. 1, colonies of the symbiotic culture prepared by the preparation method of example 1 of the present invention, cultured symbiotic culture without symbiotic medium at different time points were counted to obtain the colony counting results of lactic acid bacteria, bacillus subtilis, yeast and photosynthetic bacteria, as shown in table 1. As can be seen from the combination of Table 1 and FIG. 1, under aerobic culture conditions, yeasts, Bacillus subtilis and photosynthetic bacteria can rapidly grow and propagate; under the anaerobic condition, lactic acid bacteria can propagate in a large amount, photosynthetic bacteria and saccharomycetes can also grow and propagate in a small amount, and finally the numerical values of various bacteria reach a stable state.
The symbiotic culture medium and the symbiotic bacteria culture method of the embodiment of the invention can ensure that the bacteria grow well and the relative content of the bacteria can be kept unchanged. The symbiotic bacteria culture medium and the culture method provided by the embodiment of the invention can be well adapted to the growth of the symbiotic bacteria.
TABLE 1
Although the invention has been described in detail above with reference to a general description and specific examples, it will be apparent to one skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (9)
1. The symbiotic bacteria culture medium is characterized by comprising the following raw materials in percentage by mass: 2 to 10 percent of molasses, 0.3 to 2 percent of yeast extract, 0.1 to 1 percent of ammonium sulfate, 0.1 to 1 percent of dipotassium hydrogen phosphate, 0.2 to 1 percent of sodium chloride and the balance of water.
2. The symbiotic bacteria culture medium of claim 1, wherein,
the symbiotic bacteria culture medium comprises the following raw materials in percentage by mass: 2.2 to 9.8 percent of molasses, 0.5 to 1.8 percent of yeast extract, 0.2 to 0.8 percent of ammonium sulfate, 0.2 to 0.8 percent of dipotassium hydrogen phosphate, 0.4 to 0.8 percent of sodium chloride and the balance of water.
3. The symbiotic bacteria culture medium of claim 1, wherein,
the symbiotic bacteria culture medium comprises the following raw materials in percentage by mass: 6% of molasses, 1.2% of yeast extract, 0.5% of ammonium sulfate, 0.4% of dipotassium phosphate, 0.6% of sodium chloride and the balance of water.
4. The symbiotic bacteria culture medium according to any one of claims 1 to 3,
the pH value of the symbiotic bacteria culture medium is 7.
5. A method for culturing a symbiont comprising culturing the symbiont on the culture medium according to claim 1.
6. The method for culturing symbiotic bacteria according to claim 5, wherein the culture medium comprises a culture medium,
the symbiotic bacteria include lactic acid bacteria, bacillus subtilis, yeast and photosynthetic bacteria.
7. The method for culturing symbiotic bacteria according to claim 6,
the method comprises the following steps:
sterilizing the symbiotic bacteria culture medium at 121 ℃ for 10-20 min;
respectively adding the lactic acid bacteria, the bacillus subtilis, the saccharomycetes and the photosynthetic bacteria into the symbiotic bacteria culture medium;
keeping the temperature of the symbiotic bacteria culture medium at 28-32 ℃, firstly carrying out aerobic fermentation culture for 5-8h, then carrying out anaerobic fermentation culture for 5-8h, repeating the steps, and carrying out culture for 48-80h to obtain a symbiotic culture product.
8. The method for culturing symbiotic bacteria according to claim 7,
the aerobic fermentation culture conditions are as follows: in the symbiotic bacteria culture medium, sterile air is intermittently introduced according to the aeration ratio of 0.3-0.5, and the stirring speed is 100-150 rpm.
9. The method for culturing symbiotic bacteria according to claim 7,
the anaerobic fermentation culture conditions are as follows: the symbiotic culture medium was stopped from stirring and the aeration of air was stopped.
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