Culture medium and preparation method thereof

Abstract

The present invention provides a culture medium and a preparation method thereof, and relates to the technical field of microbial culture. The preparation method of the culture medium comprises the following steps: mixing the raw materials of the culture medium according to the formula ratio, placing in a closed container, oscillating at a constant temperature, adjusting the pH value, sterilizing, and packaging to obtain the culture medium. In the technical scheme of the present invention, by adding tannic acid to the culture medium, the antioxidant performance of the culture medium can be improved, and the generation of miscellaneous bacteria can be inhibited to a certain extent, and the quorum sensing system of subtropical saccharopolyspora can be participated in to indirectly affect the toxin production of subtropical saccharopolyspora, thereby promoting the subtropical saccharopolyspora to produce anti-Staphylococcus aureus substances when culturing subtropical saccharopolyspora, and then being used to inhibit the activity of Staphylococcus aureus.

Description

Technical Field

[0001] The present invention relates to the technical field of microorganism cultivation, and in particular to a culture medium and a configuration method thereof. Background Art

[0002] Antibiotics need to be used before, during and after the surgery to implant an artificial lens to prevent bacterial eye infection. Bacterial eye infection is caused by a variety of bacteria and is commonly treated with drugs such as levofloxacin hydrochloride eye drops, tobramycin eye drops, and neomycin sulfate eye drops. However, the bacteria gradually develop resistance to commonly used antibiotics, prolonging the treatment cycle and worsening the treatment effect.

[0003] In the prior art, antibiotic drugs are produced by culturing microorganisms. However, the initial cultivation of microorganisms is time-consuming and prone to problems such as culture medium leakage or contamination, which ultimately leads to a decrease in the survival rate of the microorganisms and a low therapeutic effect on bacterial infections. Summary of the invention

[0004] The main purpose of the present invention is to provide a culture medium and a preparation method thereof, aiming to provide a culture medium for culturing subtropical Saccharopolyspora to reduce the occurrence of bacterial infection in the eye.

[0005] The present invention provides a method for preparing a culture medium for culturing subtropical Saccharopolyspora, wherein the method comprises the following steps: Provide glucose, tannic acid, plant protein hydrolyzate, corn steep liquor, cottonseed meal, magnesium sulfate, lecithin, agar and water, mix them and place them in a sealed container, oscillate at a constant temperature, adjust the pH value, sterilize, and package them to obtain a culture medium.

[0006] In one embodiment, in the step of mixing and placing in a sealed container, the concentration of glucose is 30-50 g / L, the concentration of tannic acid is 10-20 mg / L, the concentration of plant protein hydrolysate is 10-20 g / L, the concentration of corn steep liquor is 10-20 g / L, the concentration of cottonseed powder is 10-30 g / L, the concentration of magnesium sulfate is 1-3 g / L, the concentration of lecithin is 20-100 mg / L, and the concentration of agar is 10-20 g / L.

[0007] In one embodiment, in the step of mixing and placing in a sealed container, the concentration of glucose is 40 g / L, the concentration of tannic acid is 15 mg / L, the concentration of plant protein hydrolysate is 15 g / L, the concentration of corn steep liquor is 15 g / L, the concentration of cottonseed powder is 20 g / L, the concentration of magnesium sulfate is 2 g / L, the concentration of lecithin is 50 mg / L, and the concentration of agar is 15 g / L.

[0008] In one embodiment, the temperature of the constant temperature oscillation is 60-90°C; The speed of constant temperature oscillation is 250~300 rpm; The constant temperature oscillation time is 3 to 5 minutes.

[0009] In one embodiment, the pH value is adjusted by using an acid solution or an alkali solution; in the step of adjusting the pH value, the pH value is adjusted to 7.0-7.5.

[0010] In one embodiment, the acid solution includes HCl solution, and the alkali solution includes NaOH.

[0011] In one embodiment, during the sterilization step: The culture medium raw materials are sterilized in a closed container at a temperature of 121~124 ℃ and a time of 15~30 min.

[0012] In one embodiment, in the packaging step, the sterilized culture medium is placed on an ultra-clean workbench for packaging.

[0013] In one embodiment, the culture medium is a solid culture medium.

[0014] The present invention also provides a culture medium, which is prepared according to the culture medium preparation method as described above.

[0015] The present invention also provides an application of a culture medium in the preparation of an antibacterial preparation, wherein the culture medium includes the culture medium as described above, and the antibacterial preparation is used to prevent eye infection.

[0016] In the technical scheme of the present invention, by adding tannic acid to the culture medium, the antioxidant performance of the culture medium can be improved, and the generation of miscellaneous bacteria can be inhibited to a certain extent. It can also participate in the quorum sensing system of subtropical Saccharopolyspora to indirectly affect the toxin production of subtropical Saccharopolyspora, thereby promoting the subtropical Saccharopolyspora to produce anti-Staphylococcus aureus substances when culturing subtropical Saccharopolyspora, and then used to inhibit the activity of Staphylococcus aureus; by adopting constant temperature oscillation, the dissolution of the culture medium raw materials can be accelerated. Since oscillation accelerates dissolution, the temperature setting during constant temperature oscillation is relatively low, and the pH value can be adjusted without cooling, which further accelerates the configuration process of the culture medium; by sterilizing first and then packaging, the problem of leakage or contamination of the culture medium caused by high temperature and high pressure sterilization during sterilization after packaging can be reduced. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without paying creative work.

[0018] Figure 1 It is a schematic diagram of the configuration process of the culture medium in the prior art; Figure 2 A schematic diagram of a process for configuring a culture medium provided by the present invention in one embodiment; Figure 3 It is a comparison chart of the relationship between the culture medium preparation time and the preparation batch corresponding to Example 1 of the present invention and Comparative Example 1.

[0019] The purpose, features and advantages of the present invention will be further described with reference to the accompanying drawings in conjunction with the embodiments. DETAILED DESCRIPTION

[0020] To make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them.

[0021] It should be noted that, in the embodiments, those without specifying specific conditions are carried out according to normal conditions or conditions recommended by the manufacturer. Those without specifying the manufacturer of reagents or instruments used are conventional products that can be purchased commercially. In addition, the meaning of "and / or" appearing in the full text includes three parallel schemes, and "A and / or B" is taken as an example, including schemes A, B, or A and B that meet the same time. In addition, the technical schemes between the various embodiments can be combined with each other, but must be based on the ability of ordinary technicians in the field to achieve. When the combination of the technical schemes is contradictory or cannot be achieved, it should be considered that the combination of such technical schemes does not exist, and is not within the scope of protection required by the present invention. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in the field without making creative work, all belong to the scope of protection of the present invention.

[0022] Antibiotics need to be used before, during and after the surgery to implant an artificial lens to prevent bacterial eye infection. Bacterial eye infection is caused by a variety of bacteria and is commonly treated with drugs such as levofloxacin hydrochloride eye drops, tobramycin eye drops, and neomycin sulfate eye drops. However, the bacteria gradually develop resistance to commonly used antibiotics, prolonging the treatment cycle and worsening the treatment effect.

[0023] In the prior art, antibiotic drugs are produced by culturing microorganisms. However, the initial cultivation of microorganisms is time-consuming and prone to problems such as leakage or contamination of the culture medium, which ultimately leads to a decrease in the survival rate of the microorganisms and a decrease in the therapeutic effect on bacterial infections.

[0024] In view of this, the present invention proposes a method for preparing a culture medium for culturing subtropical Saccharopolyspora. The method for preparing the culture medium comprises the following steps: providing glucose, tannic acid, plant protein hydrolyzate, corn steep liquor, cottonseed meal, magnesium sulfate, lecithin, agar and water, mixing them and placing them in a sealed container, oscillating at a constant temperature, adjusting the pH value, sterilizing, and packaging to obtain the culture medium.

[0025] In the technical scheme of the present invention, by adding tannic acid to the culture medium, the antioxidant performance of the culture medium can be improved, and the generation of miscellaneous bacteria can be inhibited to a certain extent. It can also participate in the quorum sensing system of subtropical Saccharopolyspora to indirectly affect the toxin production of subtropical Saccharopolyspora, thereby promoting the subtropical Saccharopolyspora to produce anti-Staphylococcus aureus substances when culturing subtropical Saccharopolyspora, and then used to inhibit the activity of Staphylococcus aureus; by adopting constant temperature oscillation, the dissolution of the culture medium raw materials can be accelerated. Since oscillation accelerates dissolution, the temperature setting during constant temperature oscillation is relatively low, and the pH value can be adjusted without cooling, which further accelerates the configuration process of the culture medium; by sterilizing first and then packaging, the problem of leakage or contamination of the culture medium caused by high temperature and high pressure sterilization during sterilization after packaging can be reduced.

[0026] In one embodiment, in the step of mixing and placing in a sealed container, the concentration of glucose is 30-50 g / L, the concentration of tannic acid is 10-20 mg / L, the concentration of plant protein hydrolysate is 10-20 g / L, the concentration of corn steep liquor is 10-20 g / L, the concentration of cottonseed powder is 10-30 g / L, the concentration of magnesium sulfate is 1-3 g / L, the concentration of lecithin is 20-100 mg / L, and the concentration of agar is 10-20 g / L.

[0027] It should be noted that high concentrations of tannic acid have antibacterial effects and have a certain degree of influence on the survival rate of subtropical Saccharopolyspora, while lower concentrations of tannic acid have no obvious antibacterial effect, but still have a certain antioxidant effect. In one embodiment, in the step of mixing and placing in a sealed container, the concentration of glucose is 40 g / L, the concentration of tannic acid is 15 mg / L, the concentration of plant protein hydrolysate is 15 g / L, the concentration of corn steep liquor is 15 g / L, the concentration of cottonseed powder is 20 g / L, the concentration of magnesium sulfate is 2 g / L, the concentration of lecithin is 50 mg / L, and the concentration of agar is 15 g / L.

[0028] In one embodiment, the temperature of the constant temperature oscillation is 60-90°C; The speed of constant temperature oscillation is 250~300 rpm; The constant temperature oscillation time is 3 to 5 minutes.

[0029] In the technical solution of the present invention, by adjusting the temperature and oscillation speed of the constant temperature oscillation, the dissolution of the culture medium raw materials can be accelerated, and the temperature can be controlled below 100°C, thereby reducing precipitation or condensation caused by local overheating during the dissolution process and shortening the cooling rate; by adjusting the oscillation speed to 250-300 rpm, all components are fully mixed to ensure that the final prepared culture medium has good uniformity and consistency between batches; by adjusting the oscillation time to 3-5 min, most of the components of the culture medium can be completely dissolved, reducing the time required for dissolution.

[0030] It should be noted that when adjusting the pH after constant temperature oscillation, it is not necessary to cool the culture medium before adjusting the pH. This is because the pH meter generally changes with temperature by 0.003 °C. -1 However, oscillating at 60-90 °C for 3-5 min is not enough to change the temperature of the culture medium in the container too much, so the measurement error of the pH meter can be controlled within 0.1 when adjusting the pH.

[0031] In one embodiment, the pH value is adjusted by using an acid solution or an alkali solution; in the step of adjusting the pH value, the pH value is adjusted to 7.0-7.5.

[0032] In the technical scheme of the present invention, acid or alkali solution can be used as different types of pH regulators to adjust the pH value according to different types of culture media, so that accurate adjustment of the pH value can be achieved; since the process of adjusting the pH value by acid and alkali solution is relatively direct and easy to measure, the pH value change can be monitored in real time by a pH meter or other online monitoring equipment, and automatic adjustment can be achieved in combination with an automatic control system, which not only improves work efficiency, but also reduces the possibility of human errors, ensuring the safety and accuracy of the operation; by adjusting the pH value to 7.0-7.5, the optimal pH of subtropical Saccharopolyspora can be achieved, the growth and development of subtropical Saccharopolyspora can be promoted, and the survival rate of subtropical Saccharopolyspora can be improved.

[0033] In one embodiment, the acid solution includes HCl solution, and the alkali solution includes NaOH.

[0034] In the technical solution of the present invention, HCl solution is used as the acid solution and NaOH is used as the alkali solution. Both of them are strong electrolytes and can quickly provide a large number of hydrogen ions or hydroxide ions to quickly adjust the pH. Chloride ions and sodium ions have little effect on the biological activity of microorganisms, so as to reduce the toxicity to the microbial samples and ensure the survival rate of the microbial samples.

[0035] In one embodiment, during the sterilization step: The culture medium raw materials are sterilized in a closed container at a temperature of 121~124 ℃ and a time of 15~30 min.

[0036] In the technical solution of the present invention, by placing the culture medium raw materials in a closed container for sterilization, the total mass of the culture medium in the closed container can be ensured to remain unchanged, thereby reducing the occurrence of leakage of the culture medium; by adjusting the temperature and duration of sterilization, sterilization can be achieved through high temperature. At the same time, since it is a closed container, a high-pressure environment can be formed within a certain range, thereby reducing the phenomenon of violent boiling of the liquid in the culture medium and further reducing the phenomenon of leakage of the culture medium.

[0037] In one embodiment, in the packaging step, the sterilized culture medium is placed on an ultra-clean workbench for packaging.

[0038] In the technical solution of the present invention, by placing the culture medium on a clean bench for subpackaging, the subpackaging volume is easy to be fixed, thereby avoiding the problems of insufficient subpackaging volume of the culture medium and contamination of the culture medium.

[0039] In one embodiment, the culture medium is a solid culture medium.

[0040] In the technical solution of the present invention, the solid culture medium is configured by the aforementioned configuration method, which can reduce bubbles in the solid culture medium, thereby reducing the occurrence of an unstable microbial growth environment caused by uneven culture medium, so that the cultured colonies grow stably and are easy to count.

[0041] The present invention also provides a culture medium, which is prepared according to the culture medium preparation method as described above.

[0042] Since the culture medium adopts all the technical solutions of all the above embodiments, it has at least all the beneficial effects brought by the technical solutions of the above embodiments, which will not be described one by one here.

[0043] The technical solution of the present invention is further described in detail below in conjunction with specific embodiments. It should be understood that the following embodiments are only used to explain the present invention, and are not used to limit the present invention.

[0044] Experimental Materials Subtropical Saccharopolyspora (SHMCC D58896): purchased from Ruichu Biotechnology (Jiangsu) Co., Ltd.

[0045] Staphylococcus aureus (RN4220 D25094): purchased from Shanghai Collection Biotechnology Center.

[0046] Example 1 This embodiment provides a subtropical Saccharopolyspora culture medium, and the preparation method thereof comprises the following steps: S1. Weigh 40 g of glucose, 15 mg of tannic acid, 15 g of plant protein hydrolyzate, 15 g of corn steep liquor, 20 g of cottonseed meal, 2 g of magnesium sulfate, 50 mg of lecithin, 15 g of agar and 1 L of distilled water, and mix them in a heat-resistant sealed container to obtain a premix; S2. Place the premix on a constant temperature shaker, set the temperature to 70 °C, the speed to 250 rpm, and the time to 3 min; S3. After the constant temperature oscillation is completed, the pH value in the high temperature resistant sealed container is adjusted to 7.3±0.2; S4. After adjusting the pH value, place the high temperature resistant sealed container at 121°C for high temperature sterilization for 20 min; S5. After cooling the sterilized high-temperature resistant sealed container to about 45°C, transfer it to a Class 100 clean bench for packaging to obtain a subtropical Saccharopolyspora culture medium.

[0047] Example 2 This embodiment provides a subtropical Saccharopolyspora culture medium, and the preparation method thereof comprises the following steps: S1. Weigh 30 g of glucose, 10 mg of tannic acid, 10 g of plant protein hydrolysate, 10 g of corn steep liquor, 10 g of cottonseed meal, 1 g of magnesium sulfate, 20 mg of lecithin, 10 g of agar and 1 L of distilled water, and mix them in a high temperature resistant sealed container to obtain a premix; S2. Place the premix on a constant temperature shaker, set the temperature to 60 °C, the speed to 300 rpm, and the time to 5 min; S3. After the constant temperature oscillation is completed, the pH value in the high temperature resistant sealed container is adjusted to 7.3±0.2; S4. After adjusting the pH value, place the high temperature resistant sealed container at 121°C for high temperature sterilization for 20 min; S5. After cooling the sterilized high-temperature resistant sealed container to about 45°C, transfer it to a Class 100 clean bench for packaging to obtain the subtropical Saccharopolyspora culture medium.

[0048] Example 3 This embodiment provides a subtropical Saccharopolyspora culture medium, and the preparation method thereof comprises the following steps: S1. Weigh 50 g of glucose, 20 mg of tannic acid, 20 g of plant protein hydrolyzate, 20 g of corn steep liquor, 30 g of cottonseed meal, 3 g of magnesium sulfate, 100 mg of lecithin, 20 g of agar and 1 L of distilled water, and mix them in a high temperature resistant sealed container to obtain a premix; S2. Place the premix on a constant temperature shaker, set the temperature to 90 °C, the speed to 250 rpm, and the time to 3 min; S3. After the constant temperature oscillation is completed, the pH value in the high temperature resistant sealed container is adjusted to 7.3±0.2; S4. After adjusting the pH value, place the high temperature resistant sealed container at 121°C for high temperature sterilization for 20 min; S5. After cooling the sterilized high-temperature resistant sealed container to about 45°C, transfer it to a Class 100 clean bench for packaging to obtain the subtropical Saccharopolyspora culture medium.

[0049] Comparative Example 1 This comparative example provides a subtropical Saccharopolyspora culture medium, and the preparation method thereof comprises the following steps: S10. Weigh 40 g of glucose, 15 g of vegetable protein hydrolysate, 15 g of corn steep liquor, 20 g of cottonseed meal, 2 g of magnesium sulfate, 50 mg of lecithin, 15 g of agar and 1 L of distilled water, and mix them in a heat-resistant sealed container to obtain a premix; S20. Heat the premix in a flask to 75 °C and stir for 15 min until it is completely dissolved; S30. Cool at room temperature for 15 min; S40. After cooling, adjust the pH value to 7.3±0.2; S50. After the pH value is adjusted, the solution in the flask is dispensed; S60. Place the packaged culture medium solution at 121°C for high temperature sterilization for 20 min to obtain subtropical Saccharopolyspora culture medium.

[0050] Comparative Example 2 This comparative example provides a subtropical Saccharopolyspora culture medium, and the preparation method thereof comprises the following steps: S10. Weigh 30 g of glucose, 10 mg of tannic acid, 10 g of plant protein hydrolyzate, 10 g of corn steep liquor, 10 g of cottonseed meal, 1 g of magnesium sulfate, 20 mg of lecithin, 10 g of agar and 1 L of distilled water, and mix them in a high temperature resistant sealed container to obtain a premix; S20. Heat the premix in a flask to 60 °C and stir for 15 min until it is completely dissolved; S30. Cool at room temperature for 15 min; S40. After cooling, adjust the pH value to 7.3±0.2; S50. After the pH value is adjusted, the solution in the flask is dispensed; S60. Place the packaged culture medium solution at 121°C for high temperature sterilization for 20 min to obtain subtropical Saccharopolyspora culture medium.

[0051] Comparative Example 3 This comparative example provides a subtropical Saccharopolyspora culture medium, and the preparation method thereof comprises the following steps: S10. Weigh 50 g of glucose, 20 mg of tannic acid, 20 g of plant protein hydrolyzate, 20 g of corn steep liquor, 30 g of cottonseed meal, 3 g of magnesium sulfate, 100 mg of lecithin, 20 g of agar and 1 L of distilled water, and mix them in a high temperature resistant sealed container to obtain a premix; S20. Heat the premix in a flask to 90 °C and stir for 15 min until it is completely dissolved; S30. Cool at room temperature for 15 min; S40. After cooling, adjust the pH value to 7.3±0.2; S50. After the pH value is adjusted, the solution in the flask is dispensed; S60. Place the packaged culture medium solution at 121°C for high temperature sterilization for 20 min to obtain subtropical Saccharopolyspora culture medium.

[0052] Configuration duration comparison The time comparison of preparing the culture medium in the embodiment and the comparative example is shown in Table 1, wherein n in Table 1 is the number of bottles prepared, and the linear regression equation of the preparation time and the preparation batch corresponding to the comparative example is: y=23x+45 (x is the number of bottles prepared); the linear regression equation of the preparation time and the preparation batch corresponding to the embodiment is: y=8x+50, and the relationship comparison diagram of the corresponding preparation time and the preparation batch is shown in Figure 3 shown.

[0053] Table 1 Comparison of the time for preparing the culture medium in the examples and comparative examples

[0054] Combining Table 1 and Figure 3 From the results, it can be clearly found that as the number of prepared bottles increases, the embodiment saves a lot of time compared with the comparative example.

[0055] Comparison of colony count and survival rate Comparison method: (1) Culture medium preparation: The culture medium of Examples 1 to 3 and Comparative Examples 1 to 3 were compared; (2) Preparation of bacterial solution: Take a certain amount of subtropical Saccharopolyspora, re-dissolve it, and dilute it several times to obtain a bacterial suspension, ensuring that there are no more than 100 cfu per milliliter of bacterial solution; (3) Inoculation: Pipette 1 ml of subtropical Saccharopolyspora and spread it on a plate. Prepare 2 plates for each strain. (4) Cultivation: Mix the bacterial solution and culture medium thoroughly. After the agar solidifies, turn the plate over and place it in an incubator at 30℃~35℃ for cultivation.

[0056] (5) Colony counting: Observe the growth of colonies, count all colonies growing on the plate, and then check with a 5-10x magnifying glass. Do not miss any colonies. Plates with colonies spreading and growing into pieces should not be counted and should be directly judged as unqualified.

[0057] The results of the colony counts of Examples 1 to 3 and Comparative Examples 1 to 3 are shown in Table 2, and the contamination of the culture medium is shown in Table 3.

[0058] Table 2 Results of colony counts in Examples 1 to 3 and Comparative Examples 1 to 3

[0059] From Table 2 we can see that: In the culture medium prepared by the preparation method provided in Examples 1 to 3, the survival rate of subtropical Saccharopolyspora was 78.8% to 86.7%, while in the culture medium prepared by the preparation method provided in Comparative Examples 1 to 3, the survival rate of subtropical Saccharopolyspora was 32.5% to 46.4%.

[0060] This indicates that the culture medium obtained by this scheme can improve the survival rate of subtropical Saccharopolyspora colonies to a certain extent.

[0061] Table 3 Statistical results of culture medium contamination in Examples 1 to 3 and Comparative Examples 1 to 3

[0062] According to Table 3, it can be seen that the old scheme (Comparative Examples 1 to 3) has 26.67% contamination, that is, 26.67% of the tests failed, resulting in inaccurate tests or repeated tests, and a huge waste of sample volume and time; while the new scheme (Examples 1 to 3) has no contamination, which reduces the waste of sample volume and shortens the culture cycle.

[0063] Comparison of anti-Staphylococcus aureus activity Comparison method: (1) Culture medium preparation: The culture medium of Examples 1 to 3 and Comparative Examples 1 to 3 were compared; (2) Preparation of bacterial solution: Inoculate subtropical Saccharopolyspora at an inoculation rate of 15 vol% and ferment the culture; (3) Fermentation temperature was 30°C, ventilation volume was 1:1, stirring speed was 180 rpm, and tank pressure was 0.02 MPa. Fermentation was carried out for 170 h. The fermentation broth was collected and diluted to obtain a bacterial suspension, ensuring that the bacterial suspension did not exceed 100 cfu per milliliter. (4) Prepare commercial PDA medium. Add 10 mL of bacterial suspension to the PDA medium, adjust the pH to 7.4, inoculate Staphylococcus aureus and culture at 37°C for 24 h. Calculate the total colony count of Staphylococcus aureus.

[0064] The colony count results of Staphylococcus aureus cultured using the mixed culture medium of the bacterial suspensions cultured in Examples 1 to 3 and Comparative Examples 1 to 3 are shown in Table 4.

[0065] Table 4 Colony count results of Staphylococcus aureus in Examples 1 to 3 and Comparative Examples 1 to 3

[0066] From Table 4 we can see that: The fermentation broth containing subtropical Saccharopolyspora obtained by culturing the culture medium provided in Examples 1 to 3 was added to PDA to culture Staphylococcus aureus. After 24 hours of culture, the survival rate of Staphylococcus aureus was 20.3%~34.6%, while in the culture medium prepared by the preparation method provided in Comparative Examples 1 to 3, the survival rate of Staphylococcus aureus was 54.8%~60.8%.

[0067] This indicates that the culture medium obtained by configuring this scheme can enhance the anti-Staphylococcus aureus activity produced by subtropical Saccharopolyspora, which is manifested in a decrease in the survival rate of Staphylococcus aureus.

[0068] The above description is only a preferred embodiment of the present invention, and does not limit the patent scope of the present invention. All equivalent structural changes made by using the contents of the present invention specification under the inventive concept of the present invention, or directly / indirectly applied in other related technical fields are included in the patent protection scope of the present invention.

Claims

1. A method for preparing a culture medium, characterized in that: For culturing subtropical Saccharopolyspora, the preparation method of the culture medium comprises the following steps: Provide glucose, tannic acid, plant protein hydrolyzate, corn steep liquor, cottonseed meal, magnesium sulfate, lecithin, agar and water, mix them and place them in a sealed container, oscillate at a constant temperature, adjust the pH value, sterilize, and package them to obtain a culture medium.

2. The method for preparing a culture medium according to claim 1, wherein: In the step of placing the mixture in a sealed container after mixing, the concentration of glucose is 30-50 g / L, the concentration of tannic acid is 10-20 mg / L, the concentration of plant protein hydrolysate is 10-20 g / L, the concentration of corn steep liquor is 10-20 g / L, the concentration of cottonseed powder is 10-30 g / L, the concentration of magnesium sulfate is 1-3 g / L, the concentration of lecithin is 20-100 mg / L, and the concentration of agar is 10-20 g / L.

3. The method for preparing a culture medium according to claim 2, wherein: In the step of mixing and placing in a sealed container, the concentration of glucose is 40 g / L, the concentration of tannic acid is 15 mg / L, the concentration of plant protein hydrolysate is 15 g / L, the concentration of corn steep liquor is 15 g / L, the concentration of cottonseed powder is 20 g / L, the concentration of magnesium sulfate is 2 g / L, the concentration of lecithin is 50 mg / L, and the concentration of agar is 15 g / L.

4. The method for preparing a culture medium according to claim 1, wherein: The temperature of constant temperature oscillation is 60~90℃; The speed of constant temperature oscillation is 250~300 rpm; The constant temperature oscillation time is 3 to 5 minutes.

5. The method for preparing a culture medium according to claim 1, wherein: Adjust the pH value with acid or alkali solution; In the step of adjusting the pH value, the pH value is adjusted to 7.0 to 7.

5.

6. The method for preparing a culture medium according to claim 5, wherein: The acid solution includes HCl solution, and the alkali solution includes NaOH.

7. The method for preparing a culture medium according to claim 1, wherein: During the sterilization process: The culture medium raw materials are sterilized in a closed container at a temperature of 121~124 ℃ and a time of 15~30 minutes.

8. The method for preparing a culture medium according to claim 1, wherein: In the packaging step: place the sterilized culture medium on the clean bench for packaging.

9. The method for preparing a culture medium according to claim 1, wherein: The culture medium is a solid culture medium.

10. A culture medium, characterized in that The culture medium is prepared according to the culture medium preparation method according to any one of claims 1 to 9.

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