Propionibacterium fermentation product as well as preparation method and application thereof
By using propionibacterium fermentation products in the food preservation field, combining the traditional Chinese herbal raw materials of Daqing Leaf and Houttuynia cordata and specific elution conditions, the environmental and health problems of existing food preservation agents are solved, and highly effective natural antibacterial agents are provided. It significantly improves the bactericidal ability of Penicillium.
Patent Information
- Application Number
- CN202411626744.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-14
- Publication Date
- 2025-01-07
- Estimated Expiration
- Not applicable · inactive patent
AI Technical Summary
The existing food preservatives are mainly chemical fungicides, which have environmental pollution and potential harm to human health. They have long-term use leads to microbial resistance and lack of natural antibacterial agents, especially insufficient research on Penicillium.
Using the fermentation product of propionate as an antibacterial agent, the fermentation medium is made by adding Chinese herbal raw materials such as large green leaves and Houttuynia cordata to the fermentation medium, and using the specific elution conditions of the macroporous resin column and silica gel column, a significant anti-green resistance is prepared. The fermentation product of the mold-acting propionibacterium.
It significantly improves the anti-penicillin effect, provides a natural food preservation method, avoids the pollution and drug resistance of chemical fungicides, and has important food preservation application value.
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Figure BDA0005134938730000071 
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Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of biomedicine, and in particular to a propionibacterium fermentation product and a preparation method and application thereof. Background Art
[0002] Food preservatives refer to food additives used to prevent food from spoiling due to microbial growth during storage. Most of the food preservatives currently used are chemical fungicides that are artificially synthesized; the use of chemical fungicides can easily cause chemical fungicide residues, which on the one hand pollute the environment, and on the other hand can cause potential harm to human health. In addition, long-term use of chemical fungicides can also cause microbial resistance. Therefore, the development of more types of antimicrobial agents, especially antimicrobial agents from natural sources, has broad application prospects in the field of food preservation.
[0003] Penicillium exists on the surface of many fruits, such as citrus and pears; it is the main microorganism that causes fruit rot and deterioration; therefore, developing an active ingredient with antibacterial effect against Penicillium has important application value for fruit preservation. However, there is no prior art research report on the antibacterial effect of Propionibacterium fermentation products against Penicillium. Summary of the invention
[0004] In order to overcome at least one technical problem existing in the prior art, the present invention provides a propionibacterium fermentation product and a preparation method and application thereof.
[0005] The technical solution of the present invention is as follows:
[0006] The present invention first provides a method for preparing a propionibacterium fermentation product, comprising the following steps: adding propionibacterium to a fermentation medium for fermentation; separating the fermentation liquid after the fermentation is completed, concentrating and drying the fermentation liquid to obtain a total fermentation product; taking the total fermentation product to obtain the propionibacterium fermentation product;
[0007] The fermentation medium contains Chinese herbal medicine raw materials.
[0008] The present invention provides a propionibacterium fermentation product prepared by a completely new method. Studies have shown that the propionibacterium fermentation product obtained by fermenting a fermentation medium with propionibacterium as the fermentation strain and adding Chinese herbal medicine raw materials including folium isatidis and houttuynia cordata has a good anti-penicillium effect; the anti-penicillium effect is significantly higher than that of the propionibacterium fermentation product obtained by fermenting the fermentation medium without adding the Chinese herbal medicine raw materials including folium isatidis and houttuynia cordata.
[0009] Preferably, the Chinese herbal medicine raw materials in the culture medium include Folium Isatidis and Houttuynia cordata.
[0010] Preferably, the weight ratio of Folium Isatidis to Houttuynia cordata is 1-5:1-5.
[0011] Most preferably, the weight ratio of Folium Isatidis to Houttuynia cordata is 1:1.
[0012] Preferably, the fermentation medium comprises the following components in parts by weight: 20-40 parts of Chinese herbal medicine raw materials; 5-15 parts of peptone; 5-15 parts of yeast extract powder; 2-4 parts of glucose; 0.5-2 parts of potassium dihydrogen phosphate; 0.1-1 part of magnesium sulfate; and 1000 parts of water.
[0013] Most preferably, the fermentation medium comprises the following components in parts by weight: 30 parts of Chinese herbal medicine raw materials; 10 parts of peptone; 10 parts of yeast extract powder; 3 parts of glucose; 0.5-2 parts of potassium dihydrogen phosphate; 0.1-1 part of magnesium sulfate; and 1000 parts of water.
[0014] Preferably, the weight of the added propionibacterium accounts for 1-5% of the weight of the fermentation medium;
[0015] Most preferably, the added weight of the Propionibacterium accounts for 3% of the weight of the fermentation medium.
[0016] Preferably, the fermentation is carried out at 30-40° C. for 3-7 days.
[0017] Most preferably, the fermentation is carried out at 37°C for 5 days.
[0018] Preferably, the preparation method of the propionibacterium fermentation product further comprises the following steps: placing the total fermentation product on a macroporous resin column, first eluting with an ethanol aqueous solution with a volume fraction of 42 to 45% to remove impurities; then eluting with an ethanol aqueous solution with a volume fraction of 57 to 60%, collecting the eluate eluted with the ethanol aqueous solution with a volume fraction of 57 to 60%, concentrating and drying to obtain the elution portion of the total fermentation product macroporous resin column; taking the elution portion of the total fermentation product macroporous resin column to obtain the propionibacterium fermentation product.
[0019] Most preferably, the method for preparing the propionibacterium fermentation product further comprises the following steps: placing the total fermentation product on a macroporous resin column, first eluting with a 44% by volume ethanol aqueous solution to remove impurities; then eluting with a 58% by volume ethanol aqueous solution, collecting the eluate eluted with the 58% by volume ethanol aqueous solution, concentrating and drying to obtain the elution portion of the total fermentation product macroporous resin column; taking the elution portion of the total fermentation product macroporous resin column to obtain the propionibacterium fermentation product.
[0020] The inventors found in further research that the anti-Penicillium effect of the macroporous resin column elution portion of the total fermentation product prepared by further passing the total fermentation product through the macroporous resin column elution conditions of the present invention is further significantly higher than that of the total fermentation product. In addition, the inventors also found that the macroporous resin column elution conditions of the present invention are very critical; only when the total fermentation product is further prepared under the macroporous resin column elution conditions of the present invention, the anti-Penicillium effect of the macroporous resin column elution portion of the total fermentation product can be further significantly higher than that of the total fermentation product; however, the anti-Penicillium effect of the macroporous resin column elution portion of the total fermentation product prepared under other macroporous resin column elution conditions cannot be further significantly higher than that of the total fermentation product.
[0021] Preferably, the macroporous resin column is a macroporous resin column of model HP-20.
[0022] Preferably, the preparation method of the propionibacterium fermentation product further includes the following steps: further placing the elution portion of the total fermentation product macroporous resin column on a silica gel column; first eluting with a solvent composed of cyclohexane and acetone in a volume ratio of 88 to 90:12 to 10 to remove impurities; then eluting with a solvent composed of cyclohexane and acetone in a volume ratio of 72 to 75:28 to 25, collecting the eluate eluted with the solvent composed of cyclohexane and acetone in a volume ratio of 72 to 75:28 to 25, concentrating and drying to obtain the elution portion of the total fermentation product silica gel column; taking the elution portion of the total fermentation product silica gel column to obtain the propionibacterium fermentation product.
[0023] Most preferably, the preparation method of the propionibacterium fermentation product further includes the following steps: further placing the elution portion of the total fermentation product macroporous resin column on a silica gel column; first eluting with a solvent composed of cyclohexane and acetone in a volume ratio of 89:11 to remove impurities; then eluting with a solvent composed of cyclohexane and acetone in a volume ratio of 73:27, collecting the eluate eluted with the solvent composed of cyclohexane and acetone in a volume ratio of 73:27, concentrating and drying to obtain the elution portion of the total fermentation product silica gel column; taking the elution portion of the total fermentation product silica gel column to obtain the propionibacterium fermentation product.
[0024] In further research, the inventors found that the anti-Penicillium effect of the total fermentation silica gel column elution portion obtained by further subjecting the total fermentation macroporous resin column elution portion to the silica gel column elution conditions described in the present invention is further significantly higher than the total fermentation macroporous resin column elution portion. In addition, the inventors also found that the silica gel column elution conditions of the present invention are very critical; the anti-Penicillium effect of the total fermentation macroporous resin column elution portion can only be further significantly higher than the total fermentation macroporous resin column elution portion if the total fermentation silica gel column elution portion is further prepared under the silica gel column elution conditions described in the present invention; however, the anti-Penicillium effect of the total fermentation silica gel column elution portion prepared under other silica gel column elution conditions cannot be further significantly higher than the total fermentation macroporous resin column elution portion.
[0025] The present invention also provides a propionibacterium fermentation product prepared by the above preparation method.
[0026] The present invention also provides an application of the propionibacterium fermentation product in the preparation of an antibacterial agent.
[0027] Preferably, the antibacterial agent is an antibacterial agent having an antibacterial effect on Penicillium.
[0028] Beneficial effects: The present invention provides a propionibacterium fermentation product prepared by a completely new method; studies have shown that the propionibacterium fermentation product obtained by the method of the present invention has an anti-Penicillium effect; therefore, the propionibacterium fermentation product of the present invention is used as an active ingredient to prepare an antibacterial agent with an antibacterial effect on Penicillium, and is further used as a food preservative in the field of food preservation, which has important application value. DETAILED DESCRIPTION
[0029] The present invention is further explained below in conjunction with specific examples, but the examples do not limit the present invention in any form.
[0030] Example 1 Preparation method of propionibacterium fermentation product
[0031] Propionibacterium freudenreichii powder (with a Propionibacterium freudenreichii content of 10 billion cfu / g) is added to a fermentation medium, and fermented at 37° C. for 5 days; after the fermentation is completed, the fermentation liquid is separated, and the fermentation liquid is concentrated and dried to obtain a total fermentation product; the total fermentation product is taken to obtain the Propionibacterium fermentation product;
[0032] The weight of the added Propionibacterium freudenreichii powder accounts for 3% of the weight of the fermentation medium;
[0033] The fermentation medium is mixed by the following components in parts by weight: 30 parts of Chinese herbal medicine raw materials; 10 parts of peptone; 10 parts of yeast extract powder; 3 parts of glucose; 0.5-2 parts of potassium dihydrogen phosphate; 0.1-1 parts of magnesium sulfate; 1000 parts of water;
[0034] The Chinese herbal medicine raw material is prepared by mixing and crushing Isatis indigotica and Houttuynia cordata in a weight ratio of 1:1.
[0035] Example 2 Preparation method of propionibacterium fermentation product
[0036] (1) Adding Propionibacterium freudenreichii powder (with a Propionibacterium freudenreichii content of 10 billion cfu / g) to a fermentation medium, and fermenting at 37°C for 5 days; separating the fermentation broth after the fermentation is completed, and concentrating and drying the fermentation broth to obtain a total fermentation product; wherein the weight of the Propionibacterium freudenreichii powder added accounts for 3% of the weight of the fermentation medium; the fermentation medium is composed of a mixture of the following components in parts by weight: 30 parts of Chinese herbal medicine raw materials; 10 parts of peptone; 10 parts of yeast extract powder; 3 parts of glucose; 0.5-2 parts of potassium dihydrogen phosphate; 0.1-1 parts of magnesium sulfate; and 1000 parts of water; the Chinese herbal medicine raw materials are obtained by mixing and crushing Isatis indigotica and Houttuynia cordata in a weight ratio of 1:1;
[0037] (2) taking 40 times the weight of the total fermentation product, loading it into a glass column to obtain an HP-20 macroporous resin column; loading the total fermentation product onto the HP-20 macroporous resin column, first eluting the total fermentation product with 3 times the volume of a 44% ethanol aqueous solution to remove impurities; then eluting with 5 times the column volume of a 58% ethanol aqueous solution, collecting the eluate eluted with the 58% ethanol aqueous solution, concentrating and drying it to obtain the elution portion of the total fermentation product macroporous resin column; taking the elution portion of the total fermentation product macroporous resin column to obtain the Propionibacterium fermentation product.
[0038] Example 3 Preparation method of propionibacterium fermentation product
[0039] (1) Adding Propionibacterium freudenreichii powder (with a Propionibacterium freudenreichii content of 10 billion cfu / g) to a fermentation medium, and fermenting at 37°C for 5 days; separating the fermentation broth after the fermentation is completed, and concentrating and drying the fermentation broth to obtain a total fermentation product; wherein the weight of the Propionibacterium freudenreichii powder added accounts for 3% of the weight of the fermentation medium; the fermentation medium is composed of a mixture of the following components in parts by weight: 30 parts of Chinese herbal medicine raw materials; 10 parts of peptone; 10 parts of yeast extract powder; 3 parts of glucose; 0.5-2 parts of potassium dihydrogen phosphate; 0.1-1 parts of magnesium sulfate; and 1000 parts of water; the Chinese herbal medicine raw materials are obtained by mixing and crushing Isatis indigotica and Houttuynia cordata in a weight ratio of 1:1;
[0040] (2) taking 40 times the weight of the total fermentation product, loading it into a glass column to obtain an HP-20 macroporous resin column; loading the total fermentation product onto the HP-20 macroporous resin column, first eluting the total fermentation product with 3 times the volume of a 44% ethanol aqueous solution to remove impurities; then eluting with 5 times the column volume of a 58% ethanol aqueous solution, collecting the eluate eluted with the 58% ethanol aqueous solution, concentrating and drying it to obtain the elution portion of the total fermentation product macroporous resin column;
[0041] (3) taking 30 times the weight of 200-300 mesh silica gel from the elution portion of the total fermentation macroporous resin column, loading it into a glass column to obtain a silica gel column; firstly eluting with a solvent composed of cyclohexane and acetone in a volume ratio of 89:11 for 3 times the column volume to remove impurities; then eluting with a solvent composed of cyclohexane and acetone in a volume ratio of 73:27 for 3 times the column volume, collecting the eluate eluted with the solvent composed of cyclohexane and acetone in a volume ratio of 73:27, concentrating and drying to obtain the elution portion of the total fermentation silica gel column; taking the elution portion of the total fermentation silica gel column to obtain the Propionibacterium fermentation product.
[0042] Comparative Example 1 Preparation method of propionibacterium fermentation product
[0043] Propionibacterium freudenreichii powder (with a Propionibacterium freudenreichii content of 10 billion cfu / g) is added to a fermentation medium, and fermented at 37° C. for 5 days; after the fermentation is completed, the fermentation liquid is separated, and the fermentation liquid is concentrated and dried to obtain a total fermentation product; the total fermentation product is taken to obtain the Propionibacterium fermentation product;
[0044] The weight of the added Propionibacterium freudenreichii powder accounts for 3% of the weight of the fermentation medium;
[0045] The fermentation medium is prepared by mixing the following components in parts by weight: 10 parts of peptone; 10 parts of yeast extract powder; 3 parts of glucose; 0.5-2 parts of potassium dihydrogen phosphate; 0.1-1 part of magnesium sulfate; and 1000 parts of water.
[0046] Comparative Example 2 Preparation method of propionibacterium fermentation product
[0047] (1) Adding Propionibacterium freudenreichii powder (with a Propionibacterium freudenreichii content of 10 billion cfu / g) to a fermentation medium, and fermenting at 37°C for 5 days; separating the fermentation broth after the fermentation is completed, and concentrating and drying the fermentation broth to obtain a total fermentation product; wherein the weight of the Propionibacterium freudenreichii powder added accounts for 3% of the weight of the fermentation medium; the fermentation medium is composed of a mixture of the following components in parts by weight: 30 parts of Chinese herbal medicine raw materials; 10 parts of peptone; 10 parts of yeast extract powder; 3 parts of glucose; 0.5-2 parts of potassium dihydrogen phosphate; 0.1-1 parts of magnesium sulfate; and 1000 parts of water; the Chinese herbal medicine raw materials are obtained by mixing and crushing Isatis indigotica and Houttuynia cordata in a weight ratio of 1:1;
[0048] (2) taking 40 times the weight of the total fermentation product, loading it into a glass column to obtain an HP-20 macroporous resin column; loading the total fermentation product onto the HP-20 macroporous resin column, first eluting the total fermentation product with 3 times the volume of a 20% ethanol aqueous solution to remove impurities; then eluting with 5 times the column volume of a 40% ethanol aqueous solution, collecting the eluate eluted with the 40% ethanol aqueous solution, concentrating and drying it to obtain the elution portion of the total fermentation product macroporous resin column; taking the elution portion of the total fermentation product macroporous resin column to obtain the propionibacterium fermentation product.
[0049] Comparative Example 3 Preparation method of propionibacterium fermentation product
[0050] (1) Adding Propionibacterium freudenreichii powder (with a Propionibacterium freudenreichii content of 10 billion cfu / g) to a fermentation medium, and fermenting at 37°C for 5 days; separating the fermentation broth after the fermentation is completed, and concentrating and drying the fermentation broth to obtain a total fermentation product; wherein the weight of the Propionibacterium freudenreichii powder added accounts for 3% of the weight of the fermentation medium; the fermentation medium is composed of a mixture of the following components in parts by weight: 30 parts of Chinese herbal medicine raw materials; 10 parts of peptone; 10 parts of yeast extract powder; 3 parts of glucose; 0.5-2 parts of potassium dihydrogen phosphate; 0.1-1 parts of magnesium sulfate; and 1000 parts of water; the Chinese herbal medicine raw materials are obtained by mixing and crushing Isatis indigotica and Houttuynia cordata in a weight ratio of 1:1;
[0051] (2) taking 40 times the weight of the total fermentation product, loading it into a glass column to obtain an HP-20 macroporous resin column; loading the total fermentation product onto the HP-20 macroporous resin column, first eluting the total fermentation product with 3 times the volume of a 55% ethanol aqueous solution to remove impurities; then eluting with 5 times the column volume of a 75% ethanol aqueous solution, collecting the eluate eluted with the 75% ethanol aqueous solution, concentrating and drying it to obtain the elution portion of the total fermentation product macroporous resin column; taking the elution portion of the total fermentation product macroporous resin column to obtain the Propionibacterium fermentation product.
[0052] Comparative Example 4 Preparation method of propionibacterium fermentation product
[0053] (1) Adding Propionibacterium freudenreichii powder (with a Propionibacterium freudenreichii content of 10 billion cfu / g) to a fermentation medium, and fermenting at 37°C for 5 days; separating the fermentation broth after the fermentation is completed, and concentrating and drying the fermentation broth to obtain a total fermentation product; wherein the weight of the Propionibacterium freudenreichii powder added accounts for 3% of the weight of the fermentation medium; the fermentation medium is composed of a mixture of the following components in parts by weight: 30 parts of Chinese herbal medicine raw materials; 10 parts of peptone; 10 parts of yeast extract powder; 3 parts of glucose; 0.5-2 parts of potassium dihydrogen phosphate; 0.1-1 parts of magnesium sulfate; and 1000 parts of water; the Chinese herbal medicine raw materials are obtained by mixing and crushing Isatis indigotica and Houttuynia cordata in a weight ratio of 1:1;
[0054] (2) taking 40 times the weight of the total fermentation product, loading it into a glass column to obtain an HP-20 macroporous resin column; loading the total fermentation product onto the HP-20 macroporous resin column, first eluting the total fermentation product with 3 times the volume of a 44% ethanol aqueous solution to remove impurities; then eluting with 5 times the column volume of a 58% ethanol aqueous solution, collecting the eluate eluted with the 58% ethanol aqueous solution, concentrating and drying it to obtain the elution portion of the total fermentation product macroporous resin column;
[0055] (3) taking 30 times the weight of 200-300 mesh silica gel from the elution portion of the total fermentation macroporous resin column, loading it into a glass column to obtain a silica gel column; firstly eluting with a solvent composed of cyclohexane and acetone in a volume ratio of 99:1 for 3 times the column volume to remove impurities; then eluting with a solvent composed of cyclohexane and acetone in a volume ratio of 90:10 for 3 times the column volume, collecting the eluate eluted with the solvent composed of cyclohexane and acetone in a volume ratio of 90:10, concentrating and drying to obtain the elution portion of the total fermentation silica gel column; taking the elution portion of the total fermentation silica gel column to obtain the Propionibacterium fermentation product.
[0056] Comparative Example 5 Preparation method of propionibacterium fermentation product
[0057] (1) Adding Propionibacterium freudenreichii powder (with a Propionibacterium freudenreichii content of 10 billion cfu / g) to a fermentation medium, and fermenting at 37°C for 5 days; separating the fermentation broth after the fermentation is completed, and concentrating and drying the fermentation broth to obtain a total fermentation product; wherein the weight of the Propionibacterium freudenreichii powder added accounts for 3% of the weight of the fermentation medium; the fermentation medium is composed of a mixture of the following components in parts by weight: 30 parts of Chinese herbal medicine raw materials; 10 parts of peptone; 10 parts of yeast extract powder; 3 parts of glucose; 0.5-2 parts of potassium dihydrogen phosphate; 0.1-1 parts of magnesium sulfate; and 1000 parts of water; the Chinese herbal medicine raw materials are obtained by mixing and crushing Isatis indigotica and Houttuynia cordata in a weight ratio of 1:1;
[0058] (2) taking 40 times the weight of the total fermentation product, loading it into a glass column to obtain an HP-20 macroporous resin column; loading the total fermentation product onto the HP-20 macroporous resin column, first eluting the total fermentation product with 3 times the volume of a 44% ethanol aqueous solution to remove impurities; then eluting with 5 times the column volume of a 58% ethanol aqueous solution, collecting the eluate eluted with the 58% ethanol aqueous solution, concentrating and drying it to obtain the elution portion of the total fermentation product macroporous resin column;
[0059] (3) taking 30 times the weight of 200-300 mesh silica gel from the elution portion of the total fermentation macroporous resin column, loading it into a glass column to obtain a silica gel column; firstly eluting with a solvent composed of cyclohexane and acetone in a volume ratio of 70:30 for 3 times the column volume to remove impurities; then eluting with a solvent composed of cyclohexane and acetone in a volume ratio of 50:50 for 3 times the column volume, collecting the eluate eluted with the solvent composed of cyclohexane and acetone in a volume ratio of 50:50, concentrating and drying to obtain the elution portion of the total fermentation silica gel column; taking the elution portion of the total fermentation silica gel column to obtain the Propionibacterium fermentation product.
[0060] Experimental Example 1 Anti-Penicillium Experiment of Propionibacterium Fermentation Product of the Present Invention
[0061] The propionibacterium fermentation products to be tested were prepared into sample solutions with concentrations of 100, 50, 25, 12.5, 6.25, 3.125, 1.563, 0.781, 0.391, and 0.195 μg / mL. The propionibacterium to be tested were the propionibacterium fermentation products prepared in Examples 1 to 3 and Comparative Examples 1 to 5, respectively.
[0062] Take 50 μL of 10 8 cfu / mL of Penicillium citrinum bacterial solution was spread on the agar medium plate, and then 50 μL of the test sample solution with different concentrations was evenly spread on it; finally, it was cultured in a constant temperature incubator at 37°C for 24 hours. After the culture was completed, the growth of the strain on the agar medium plate was observed. Each group of experiments was repeated 3 times; the concentration with colony count less than 5 was taken as the minimum bactericidal concentration (MBC); the test results are shown in Table 1.
[0063] Table 1. Minimum bactericidal concentration (MBC) of Propionibacterium fermentation products
[0064]
[0065]
[0066] It can be seen from the experimental results in Table 1 that the minimum bactericidal concentration (MBC) of the Propionibacterium fermentation product prepared in Example 1 is significantly lower than that of the Propionibacterium fermentation product prepared in Comparative Example 1; this indicates that: using Propionibacterium as the fermentation strain, the Propionibacterium fermentation product obtained by fermenting with Chinese herbal raw materials including Folium Isatidis and Houttuynia cordata in the fermentation medium has a good anti-Penicillium effect; its anti-Penicillium effect is significantly higher than that of the Propionibacterium fermentation product obtained by fermenting with no Chinese herbal raw materials including Folium Isatidis and Houttuynia cordata in the fermentation medium.
[0067] It can be seen from the experimental results in Table 1 that the minimum bactericidal concentration (MBC) of the Propionibacterium fermentation product prepared in Example 2 is much lower than that of the Propionibacterium fermentation product prepared in Example 1; this indicates that the anti-Penicillium effect of the macroporous resin column elution portion of the total fermentation product prepared by further subjecting the total fermentation product to the macroporous resin column elution conditions of the present invention is further much higher than that of the total fermentation product.
[0068] It can be seen from the experimental results in Table 1 that the minimum bactericidal concentration (MBC) of the Propionibacterium fermentation products prepared in Comparative Examples 2 and 3 is much lower than that of the Propionibacterium fermentation product prepared in Example 1, which is much lower than that of the Propionibacterium fermentation product prepared in Example 2; this shows that: the elution conditions of the macroporous resin column of the present invention are very critical; only when the total fermentation product is further prepared under the macroporous resin column elution conditions of the present invention, the anti-Penicillium effect of the macroporous resin column elution part of the total fermentation product can be further significantly higher than that of the total fermentation product; however, the anti-Penicillium effect of the macroporous resin column elution part of the total fermentation product prepared under other macroporous resin column elution conditions cannot be further significantly higher than that of the total fermentation product.
[0069] It can be seen from the experimental results in Table 1 that the minimum bactericidal concentration (MBC) of the Propionibacterium fermentation product prepared in Example 3 is further significantly lower than that of the Propionibacterium fermentation product prepared in Example 2; this indicates that the anti-Penicillium effect of the total fermentation silica gel column elution portion obtained by further subjecting the total fermentation macroporous resin column elution portion to the silica gel column elution conditions described in the present invention is further significantly higher than that of the total fermentation macroporous resin column elution portion.
[0070] It can be seen from the experimental results in Table 1 that the minimum bactericidal concentration (MBC) of the Propionibacterium fermentation product prepared in Comparative Example 4 is much lower than that of the Propionibacterium fermentation product prepared in Example 2, and the minimum bactericidal concentration (MBC) of the Propionibacterium fermentation product prepared in Example 3 is not reduced compared with that of the Propionibacterium fermentation product prepared in Example 2. This shows that the elution conditions of the silica gel column of the present invention are very critical; the anti-penicillium effect of the total fermentation product macroporous resin column elution site can only be further significantly higher than the total fermentation product macroporous resin column elution site if the total fermentation product silica gel column elution site is further prepared under the silica gel column elution conditions of the present invention; however, the anti-penicillium effect of the total fermentation product silica gel column elution site prepared under other silica gel column elution conditions cannot be further higher or significantly higher than the total fermentation product macroporous resin column elution site.
Claims
1. A method for preparing a propionibacterium fermentation product, comprising the following steps: adding propionibacterium to a fermentation medium for fermentation; separating the fermentation liquid after the fermentation is completed, concentrating and drying the fermentation liquid to obtain a total fermentation product; taking the total fermentation product to obtain the propionibacterium fermentation product; It is characterized in that The fermentation medium contains Chinese herbal medicine raw materials.
2. The method for preparing a propionibacterium fermentation product according to claim 1, characterized in that: The Chinese herbal raw materials in the culture medium include Folium Isatidis and Houttuynia cordata.
3. The method for preparing a propionibacterium fermentation product according to claim 2, characterized in that: The weight ratio of Isatis indigotica to Houttuynia cordata is 1-5:1-5. Most preferably, the weight ratio of Folium Isatidis to Houttuynia cordata is 1:
1.
4. The method for preparing a propionibacterium fermentation product according to claim 1, characterized in that: The fermentation medium comprises the following components in parts by weight: 20-40 parts of Chinese herbal medicine raw materials; 5-15 parts of peptone; 5-15 parts of yeast extract powder; 2-4 parts of glucose; 0.5-2 parts of potassium dihydrogen phosphate; 0.1-1 parts of magnesium sulfate; and 1000 parts of water.
5. The method for preparing a propionibacterium fermentation product according to claim 4, characterized in that: The fermentation medium comprises the following components in parts by weight: 30 parts of Chinese herbal medicine raw materials; 10 parts of peptone; 10 parts of yeast extract powder; 3 parts of glucose; 0.5-2 parts of potassium dihydrogen phosphate; 0.1-1 part of magnesium sulfate; and 1000 parts of water.
6. The method for preparing a propionibacterium fermentation product according to claim 1, characterized in that: The fermentation is carried out at 30-40°C for 3-7 days. Most preferably, the fermentation is carried out at 37°C for 5 days.
7. The method for preparing a propionibacterium fermentation product according to claim 1, characterized in that: The method further comprises the following steps: The total fermentation product is placed on a macroporous resin column, and impurities are first eluted with an ethanol aqueous solution with a volume fraction of 42-45%; then, it is eluted with an ethanol aqueous solution with a volume fraction of 57-60%, and the eluate eluted with the ethanol aqueous solution with a volume fraction of 57-60% is collected, concentrated and dried to obtain the elution portion of the total fermentation product macroporous resin column; the elution portion of the total fermentation product macroporous resin column is taken to obtain the propionibacterium fermentation product.
8. The method for preparing a propionibacterium fermentation product according to claim 1, characterized in that: The method further comprises the following steps: The total fermentation product is placed on a macroporous resin column, and impurities are first eluted with an ethanol aqueous solution with a volume fraction of 44%; then it is eluted with an ethanol aqueous solution with a volume fraction of 58%, and the eluate eluted with the ethanol aqueous solution with a volume fraction of 58% is collected, concentrated and dried to obtain the elution portion of the total fermentation product macroporous resin column; the elution portion of the total fermentation product macroporous resin column is taken to obtain the propionibacterium fermentation product.
9. The propionibacterium fermentation product obtained by the preparation method according to any one of claims 1 to 8.
10. Use of the propionibacterium fermentation product according to claim 9 in the preparation of an antibacterial agent.