Fermentation method for increasing fermentation yield of monensin and application of fermentation method

Abstract

The invention relates to a fermentation method for increasing the fermentation yield of monensin and application of the fermentation method. The fermentation method comprises the following steps: inoculating streptomyces cinnamomi into a seed culture medium for seed culture; inoculating the seed culture solution into a fermentation culture medium for fermentation culture to obtain monensin; the formula of the fermentation culture medium comprises the following components: soybean oil, methyl oleate, glucose, low-temperature soybean meal, Na2SO4, Al2 (SO4) 3, ferrous sulfate, manganese chloride, dipotassium phosphate, CaCO3 and L-ascorbic acid, and the pH value of the fermentation culture medium is 7.0-7.5. According to the method, by optimizing the formula of the culture medium, the yield of the monensin is greatly increased, the fermentation period is greatly shortened, the cost is greatly saved, the fermentation efficiency is improved, and the method is a monensin fermentation production technology which is high in yield, cost-saving and synergistic.

Description

Technical Field

[0001] The present invention belongs to the technical field of the fermentation production process of monensin, and relates to a fermentation method for improving the fermentation yield of monensin and its application. Background Art

[0002] Monensin is a class of polyether ionophore antibiotics produced by Streptomyces cinnamonensis, and is widely used in the prevention and treatment of coccidiosis in chickens, lambs, calves, rabbits and the promotion of the growth of ruminants. Monensin has the effects of controlling the proportion of volatile fatty acids in the rumen, reducing the degradation of proteins in the rumen, reducing the dry matter consumption of feed, improving the utilization rate of nutrients and increasing the energy utilization rate of animals. In recent years, due to its good inhibitory activity against various types of tumor cells in in vitro experiments, monensin is also considered a potential anti-tumor agent.

[0003] At present, the method for obtaining monensin industrially is mainly to culture Streptomyces cinnamonensis, collect the fermentation broth and purify its fermentation products. However, the traditional fermentation method often has the following disadvantages: the fermentation time is relatively long and the yield is relatively low, which increases the industrial production cost of monensin, affects the industrialization efficiency of monensin, and at the same time, the growth state of the strain is unstable, and the phenomenon of cell adhesion is serious, which affects the fermentation effect of the strain. Therefore, it is very meaningful to develop a fermentation method that can reduce the production cycle and increase the yield of monensin. Summary of the Invention

[0004] Aiming at the deficiencies of the prior art, the purpose of the present invention is to provide a fermentation method for improving the fermentation yield of monensin and its application.

[0005] To achieve the purpose of the present invention, the following technical solutions are adopted:

[0006] In the first aspect, the present invention provides a fermentation method for improving the fermentation yield of monensin, and the fermentation method comprises the following steps:

[0007] (1) Take Streptomyces cinnamonensis and inoculate it into a seed culture medium for seed culture;

[0008] (2) Take the seed culture solution and inoculate it into a fermentation culture medium for fermentation culture to obtain monensin;

[0009] The formula of the fermentation medium per 100 mL by volume comprises the following components: 1 - 5 mL of soybean oil, 0.5 - 2 mL of methyl oleate, 1 - 3 g of glucose, 2 - 5 g of low-temperature soybean powder, 0.1 - 0.4 g of Na2SO4, 0.05 - 0.1 g of Al2(SO4)3, 0.005 - 0.015 g of ferrous sulfate, 0.01 - 0.05 g of manganese chloride, 0.005 - 0.01 g of dipotassium hydrogen phosphate, 0.1 - 0.5 g of CaCO3, 0.001 - 0.003 g of L-ascorbic acid, and pH = 7.0 - 7.5.

[0010] The present invention develops a new fermentation medium formula suitable for monensin, which can significantly improve the fermentation yield of monensin and shorten the production cycle.

[0011] The volume of the above soybean oil can be selected as 1 mL, 1.5 mL, 2 mL, 2.5 mL, 3 mL, 3.5 mL, 4 mL, 4.5 mL, 5 mL, etc.; the volume of the above methyl oleate can be selected as 0.5 mL, 0.8 mL, 1 mL, 1.2 mL, 1.4 mL, 1.5 mL, 1.6 mL, 1.8 mL, 2 mL, etc.; the mass of the above glucose can be selected as 1 g, 1.2 g, 1.5 g, 1.8 g, 2 g, 2.2 g, 2.5 g, 2.8 g, 3 g, etc.; the mass of the above low-temperature soybean powder can be selected as 2 g, 2.2 g, 2.5 g, 2.8 g, 3 g, 3.2 g, 3.5 g, 3.8 g, 4 g, 4.2 g, 4.5 g, 4.8 g, 5 g, etc.; the mass of the above Na2SO4 can be selected as 0.1 g, 0.15 g, 0.2 g, 0.25 g, 0.3 g, 0.35 g, 0.4 g, etc.; the mass of the above Al2(SO4)3 can be selected as 0.05 g, 0.06 g, 0.07 g, 0.08 g, 0.09 g, 0.1 g, etc.; the mass of the above ferrous sulfate can be selected as 0.005 g, 0.007 g, 0.008 g, 0.01 g, 0.012 g, 0.013 g, 0.015 g, etc.; the mass of the above manganese chloride can be selected as 0.01 g, 0.015 g, 0.02 g, 0.025 g, 0.03 g, 0.035 g, 0.04 g, 0.045 g, 0.05 g, etc.; the mass of the above dipotassium hydrogen phosphate can be selected as 0.005 g, 0.006 g, 0.007 g, 0.008 g, 0.009 g, 0.01 g, etc.; the mass of the above CaCO3 can be selected as 0.1 g, 0.15 g, 0.2 g, 0.25 g, 0.3 g, 0.35 g, 0.4 g, 0.45 g, 0.5 g, etc.; the mass of the above L-ascorbic acid can be selected as 0.001 g, 0.0015 g, 0.002 g, 0.0025 g, 0.003 g, etc.; the above pH value can be selected as 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, etc.; other specific point values within the above numerical ranges can all be selected and will not be elaborated one by one here.

[0012] Preferably, the formula of the seed culture medium includes the following components in a volume of 100 mL: dextrin 1 - 3 g, soybean powder 0.5 - 2.5 g, yeast extract 0.05 - 0.25 g, glucose 0.3 - 0.7 g, CaCO3 0.05 - 0.15 g, pH = 7.0 - 7.5.

[0013] Furthermore, the above-optimized formula of the seed culture medium can further improve the fermentation yield of monensin and shorten the production cycle.

[0014] The mass of the above dextrin can be selected as 1 g, 1.2 g, 1.5 g, 1.8 g, 2 g, 2.2 g, 2.5 g, 2.8 g, 3 g, etc.; the mass of the above soybean powder can be selected as 0.5 g, 0.7 g, 0.8 g, 1 g, 1.2 g, 1.5 g, 1.7 g, 2 g, 2.2 g, 2.3 g, 2.5 g, etc.; the mass of the above yeast extract can be selected as 0.05 g, 0.06 g, 0.07 g, 0.08 g, 0.09 g, 0.1 g, 0.12 g, 0.15 g, 0.18 g, 0.2 g, 0.22 g, 0.25 g, etc.; the mass of the above glucose can be selected as 0.3 g, 0.35 g, 0.4 g, 0.45 g, 0.5 g, 0.55 g, 0.6 g, 0.65 g, 0.7 g, etc.; the mass of the above CaCO3 can be selected as 0.05 g, 0.07 g, 0.08 g, 0.1 g, 0.12 g, 0.13 g, 0.15 g, etc.; the above pH value can be selected as 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, etc.; other specific point values within the above numerical ranges can all be selected and will not be elaborated one by one here.

[0015] Preferably, the inoculation amount of the seed culture solution is 5-15%, such as 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, etc.

[0016] Preferably, the fermentation culture is carried out in a shaker at 100-400 rpm, such as 100 rpm, 150 rpm, 200 rpm, 250 rpm, 300 rpm, 350 rpm, 400 rpm, etc.

[0017] Other specific point values within the above numerical ranges can all be selected and will not be elaborated one by one here.

[0018] Preferably, the temperature of the fermentation culture is 35-40 °C, such as 35 °C, 36 °C, 37 °C, 38 °C, 39 °C, 40 °C, etc.

[0019] The present invention also creatively discovers that when the fermentation temperature is in the range of 35-40 °C, the effect of improving the fermentation yield of monensin and shortening the production cycle is better.

[0020] Preferably, the time of the fermentation culture is 72-120 h, such as 72 h, 76 h, 80 h, 84 h, 88 h, 92 h, 96 h, 100 h, 108 h, 112 h, 116 h, 120 h, etc.

[0021] Other specific point values within the above numerical ranges can all be selected and will not be elaborated one by one here.

[0022] The fermentation method involved in the present invention significantly shortens the fermentation cycle, reaching the plateau phase within 72 - 120 h, and reduces the production process cost.

[0023] Preferably, the temperature for seed culture is 28 - 32 °C, such as 28 °C, 29 °C, 30 °C, 31 °C, 32 °C, etc.

[0024] Preferably, the time for seed culture is 24 - 72 h, such as 24 h, 28 h, 32 h, 36 h, 40 h, 44 h, 48 h, 52 h, 56 h, 60 h, 64 h, 68 h, 72 h, etc.

[0025] Other specific point values within the above numerical ranges can be selected and will not be elaborated one by one here.

[0026] Preferably, the seed culture is carried out in a shaker at 100 - 400 rpm, such as 100 rpm, 150 rpm, 200 rpm, 250 rpm, 300 rpm, 350 rpm, 400 rpm, etc.

[0027] Preferably, the liquid loading volume of the fermentation medium is 1 / 6 - 1 / 3 of the volume of the culture container, such as 1 / 6, 1 / 5, 1 / 4, 1 / 3, etc.

[0028] The present invention also creatively discovers that by adjusting the liquid loading volume of the fermentation medium, the fermentation effect of Streptomyces cinnamonensis can also be significantly affected. When the liquid loading volume is controlled within 1 / 6 - 1 / 3 of the volume of the culture container, the production of monensin can be significantly increased.

[0029] Preferably, the liquid loading volume of the seed medium is 1 / 12 - 1 / 8 of the volume of the culture container, such as 1 / 12, 1 / 11, 1 / 10, 1 / 9, 1 / 8, etc.

[0030] Other specific point values within the above numerical ranges can be selected and will not be elaborated one by one here.

[0031] Preferably, before inoculating Streptomyces cinnamonensis into the seed medium, solid culture is first carried out: the strain is spread on the solid medium and cultured at 28 - 32 °C (such as 28 °C, 29 °C, 30 °C, 31 °C, 32 °C, etc.) for 24 - 72 h (such as 24 h, 28 h, 32 h, 36 h, 40 h, 44 h, 48 h, 52 h, 56 h, 60 h, 64 h, 68 h, 72 h, etc.).

[0032] In the second aspect, the present invention provides the application of the fermentation method for increasing the fermentation yield of monensin according to the first aspect in the preparation of monensin.

[0033] Compared with the prior art, the present invention has the following beneficial effects:

[0034] The present invention has developed a brand-new fermentation method for producing monensin. By optimizing the formula of the culture medium, the yield of monensin has been significantly increased, the fermentation cycle has been greatly shortened, the cost has been saved to a great extent, and the fermentation efficiency has been improved. It is a high-yield, cost-saving, and efficiency-enhancing monensin fermentation production technology, which is of great significance for realizing the industrialization development of veterinary antibiotics in China, expanding the scope of use, and promoting the sustainable development of the livestock industry. Detailed implementation manners

[0035] To further elaborate on the technical means and effects adopted by the present invention, the following further illustrates the technical solution of the present invention in combination with the preferred embodiments of the present invention. However, the present invention is not limited to the scope of the embodiments.

[0036] In the following implementation cases, Streptomyces cinnamonensis ATCC 15413 is used as the starting strain for monensin fermentation. This strain is a publicly available strain. In 2021, this strain was reclassified under the catalog of Streptomyces virginiae and renamed Streptomyces virginiae.

[0037] Example 1

[0038] This example provides a fermentation method for producing monensin, which is as follows:

[0039] (1) In a sterile condition, inoculate 30 μL of the original spores of the commercially available strain Streptomyces cinnamonensis ATCC 15413 onto the SFM solid medium (20 g / L soybean cake powder, 20 g / L mannitol, pH 7.0 - 7.5), leaving a distance of about 0.5 - 1 cm at the edge without spreading the bacteria; culture in an incubator at 30°C for 8 days;

[0040] (2) Take out the spore-producing plate, gently collect the fresh spores of Streptomyces cinnamonensis ATCC 15413 with a sterile cotton swab, and store them in 20% glycerol;

[0041] (3) Aspirate 30 μL of the spore suspension and inoculate and spread it on a fresh SFM solid medium, and culture in an incubator at 30°C for 48 h;

[0042] (4) Cut off about 1 cm with a sterile pipette tip 2An agar block containing hyphae was inoculated into a shake flask with a seed medium volume of 25 mL / 250 mL under aseptic conditions and cultured on a rotary shaker at 30 °C and 220 rpm for 48 h. The formula of the seed medium was (g / 100 ml): dextrin 2.0, soybean powder 1.5, yeast extract 0.15, glucose 0.5, CaCO3 0.1, pH = 7.4;

[0043] (5) According to an inoculation amount of 10% by volume of the liquid medium, the seed medium was sucked into the shake flask fermentation medium with a volume of 50 mL / 250 mL and cultured on a rotary shaker at 37 °C and 220 rpm for 96 h. Among them, the formula of the fermentation medium was (g / 100 ml): soybean oil 3.0 (mL), methyl oleate 1.0 (mL), glucose 2, low-temperature soybean powder 3.26, Na2SO4 0.22, Al2(SO4)3 0.0705, ferrous sulfate 0.011, manganese chloride 0.033, dipotassium hydrogen phosphate 0.0075, CaCO3 0.2, L-ascorbic acid 0.0019, pH 7.4.

[0044] Example 2

[0045] This example provides a fermentation method for producing monensin, which is as follows:

[0046] (1) 30 μL of the original spores of the commercially available strain Streptomyces cinnamonensis ATCC 15413 were inoculated into an SFM solid medium (20 g / L soybean cake powder, 20 g / L mannitol, pH 7.0 - 7.5) under aseptic conditions, and a distance of about 0.5 - 1 cm was left uncoated at the edge; it was cultured in an incubator at 30 °C for 8 days;

[0047] (2) After taking out the spore-producing plate, the fresh spores of Streptomyces cinnamonensis ATCC 15413 were gently collected with a sterile cotton swab and stored in 20% glycerol;

[0048] (3) 30 μL of the spore suspension was sucked and spread on a fresh SFM solid medium and cultured in an incubator at 30 °C for 48 h;

[0049] (4) Cut off an agar block containing hyphae about 1 cm with a sterile pipette tip 2 An agar block containing hyphae was inoculated into a shake flask with a seed medium volume of 30 mL / 250 mL under aseptic conditions and cultured on a rotary shaker at 28 °C and 220 rpm for 72 h. The formula of the seed medium was (g / 100 ml): dextrin 1.5, soybean powder 2, yeast extract 0.2, glucose 0.4, CaCO3 0.15, pH = 7.2;

[0050] (5) With an inoculum size of 10% by volume of the liquid medium, suck the seed medium into the shake flask fermentation medium, with a filling volume of 60 mL / 250 mL, and culture it on a rotary shaker at 38 °C and 220 rpm for 90 h. Among them, the formula of the fermentation medium is (g / 100 ml): soybean oil 2.0 (mL), methyl oleate 2.0 (mL), glucose 1.5, low-temperature soybean powder 4.2, Na2SO4 0.22, Al2(SO4)3 0.0705, ferrous sulfate 0.011, manganese chloride 0.033, dipotassium hydrogen phosphate 0.0075, CaCO3 0.2, L-ascorbic acid 0.0019, pH 7.2.

[0051] Example 3

[0052] This example provides a fermentation method for producing monensin, which is specifically as follows:

[0053] (1) Under sterile conditions, inoculate 30 μL of the original spores of the commercially available strain Streptomyces cinnamonensis ATCC 15413 on the SFM solid medium (20 g / L soybean cake powder, 20 g / L mannitol, pH 7.0 - 7.5), leaving a distance of about 0.5 - 1 cm at the edge without spreading the bacteria; culture it in an incubator at 30 °C for 8 days;

[0054] (2) Take out the spore-producing plate, gently collect the fresh spores of Streptomyces cinnamonensis ATCC 15413 with a sterile cotton swab, and store them in 20% glycerol;

[0055] (3) Suck 30 μL of the spore suspension and inoculate and spread it on a fresh SFM solid medium, and culture it in an incubator at 30 °C for 48 h;

[0056] (4) Use a sterile pipette tip to cut about 1 cm 2 agar block containing hyphae, and under sterile conditions, inoculate it into a shake flask with a seed medium with a filling volume of 22 mL / 250 mL, and culture it on a rotary shaker at 31 °C and 220 rpm for 48 h. Among them, the formula of the seed medium is (g / 100 ml): dextrin 2.5, soybean powder 1, yeast extract 0.1, glucose 0.55, CaCO3 0.08, pH = 7.5;

[0057] (5) With an inoculum size of 10% by volume of the liquid medium, aspirate the seed medium into the shake flask fermentation medium, with a filling volume of 45 mL / 250 mL, and culture it on a rotary shaker at 36 °C and 220 rpm for 102 h. Among them, the formula of the fermentation medium is (g / 100 ml): soybean oil 3.0 (mL), methyl oleate 1.0 (mL), glucose 2.5, low-temperature soybean flour 2.5, Na2SO4 0.22, Al2(SO4)3 0.0705, ferrous sulfate 0.011, manganese chloride 0.033, dipotassium hydrogen phosphate 0.0075, CaCO3 0.2, L-ascorbic acid 0.0019, pH 7.5.

[0058] Example 4

[0059] This example provides a fermentation method for producing monensin. The difference in its operation from Example 1 is only that: by adjusting the CaCO3 concentration to 0, the pH value of the seed medium is 6.8. Other conditions remain unchanged.

[0060] Example 5

[0061] This example provides a fermentation method for producing monensin. The difference in its operation from Example 1 is only that: the filling volume of the seed medium is 50 mL / 250 mL. Other conditions remain unchanged.

[0062] Example 6

[0063] This example provides a fermentation method for producing monensin. The difference in its operation from Example 1 is only that: the filling volume of the seed medium is 85 mL / 250 mL. Other conditions remain unchanged.

[0064] Example 7

[0065] This example provides a fermentation method for producing monensin. The difference in its operation from Example 1 is only that: the fermentation culture is carried out at 30 °C. Other conditions remain unchanged.

[0066] Example 8

[0067] This example provides a fermentation method for producing monensin. The difference in its operation from Example 1 is only that: the fermentation culture time is 120 h. Other conditions remain unchanged.

[0068] Example 9

[0069] This example provides a fermentation method for producing monensin. The difference in operation from Example 1 is only that: the content of the formula components in the fermentation medium is different. Among them, the formula of the fermentation medium is (g / 100ml): soybean oil 2.0 (mL), methyl oleate 2.0 (mL), glucose 1.5, low-temperature soybean meal 4.2, Na2SO4 0.22, Al2(SO4)3 0.0705, ferrous sulfate 0.011, manganese chloride 0.033, dipotassium hydrogen phosphate 0.0075, pH 7.4.

[0070] Comparative Example 1

[0071] This comparative example provides a fermentation method for producing monensin. The difference in operation from Example 1 is only that: methyl oleate is missing in the fermentation medium. Other conditions remain unchanged.

[0072] Comparative Example 2

[0073] This comparative example provides a fermentation method for producing monensin. The difference in operation from Example 1 is only that: soybean oil and methyl oleate are missing in the fermentation medium. Other conditions remain unchanged.

[0074] Comparative Example 3

[0075] This comparative example provides a fermentation method for producing monensin. The difference in operation from Example 1 is only that: low-temperature soybean meal is missing in the fermentation medium. Other conditions remain unchanged.

[0076] Comparative Example 4

[0077] This comparative example provides a fermentation method for producing monensin. The difference in operation from Example 1 is only that: glucose is missing in the fermentation medium. Other conditions remain unchanged.

[0078] Test Example

[0079] Quantitative detection of monensin:

[0080] After fermentation, 100 μL of the fermentation broth prepared in Examples 1-9 and Comparative Examples 1-4 was taken, 900 μL of methanol was added, and after mixing, it was sonicated for 30 min. After centrifugation at 12000 rpm for 1 min, the supernatant was filtered through a 0.22 μm filter membrane. After filtration, it was diluted 10 times with methanol, that is, a total dilution of 100 times.

[0081] The processed fermentation broth of each group was analyzed using an Agilent 6470 triple quadrupole liquid chromatography-mass spectrometry system equipped with a C18 chromatographic column. The mobile phase was 5% A (0.1% formic acid in water) / 95% B (acetonitrile). Isocratic elution was performed at a flow rate of 0.3 mL / min for 10 min / sample. The mass spectrometry detection mode was the MRM negative mode, and the parent / daughter ion pair was 669 / 637. The results are shown in Table 1.

[0082] Table 1

[0083]

[0084]

[0085] As can be seen from the data in Table 1, the fermentation method for producing monensin involved in the present invention can result in a high yield of monensin, a short fermentation period, greatly saving costs and improving fermentation efficiency.

[0086] The applicant declares that the present invention uses the above embodiments to illustrate the technical solutions of the present invention, but the present invention is not limited to the above embodiments, that is, it does not mean that the present invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvement to the present invention, the equivalent substitution of each raw material of the product of the present invention, the addition of auxiliary components, the selection of specific methods, etc., all fall within the protection scope and the disclosure scope of the present invention.

[0087] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept scope of the present invention, various simple modifications can be made to the technical solutions of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0088] In addition, it should be noted that, in the case of no contradiction, the various specific technical features described in the above specific embodiments can be combined in any suitable way. To avoid unnecessary repetition, the present invention will not separately describe various possible combination methods.

Claims

1. A fermentation method for increasing the fermentation yield of monensin, characterized in that, The fermentation method comprises the following steps: (1) Taking Streptomyces cinnamonensis and inoculating it into a seed medium for seed culture; (2) Taking the seed culture solution and inoculating it into a fermentation medium for fermentation culture to obtain monensin; The formula of the fermentation medium, based on a volume of 100 mL, comprises the following components: 1 - 5 mL of soybean oil, 0.5 - 2 mL of methyl oleate, 1 - 3 g of glucose, 2 - 5 g of low-temperature soybean flour, 0.1 - 0.4 g of Na2SO4, 0.05 - 0.1 g of Al2(SO4)3, 0.005 - 0.015 g of ferrous sulfate, 0.01 - 0.05 g of manganese chloride, 0.005 - 0.01 g of dipotassium hydrogen phosphate, 0.1 - 0.5 g of CaCO3, 0.001 - 0.003 g of L-ascorbic acid, pH = 7.0 - 7.

5.

2. The fermentation method for increasing the fermentation yield of monensin according to claim 1, characterized in that, The formula of the seed medium, based on a volume of 100 mL, comprises the following components: 1 - 3 g of dextrin, 0.5 - 2.5 g of soybean flour, 0.05 - 0.25 g of yeast extract, 0.3 - 0.7 g of glucose, 0.05 - 0.15 g of CaCO3, pH = 7.0 - 7.

5.

3. The fermentation method for increasing the fermentation yield of monensin according to claim 1 or 2, characterized in that, The inoculation amount of the seed culture solution is 5 - 15%; Preferably, the fermentation culture is carried out in a shaker at 100 - 400 rpm.

4. The fermentation method for increasing the fermentation yield of monensin according to any one of claims 1-3, characterized in that, The temperature of the fermentation culture is 35 - 40 °C; Preferably, the time of the fermentation culture is 72 - 120 h.

5. The fermentation method for increasing the fermentation yield of monensin according to any one of claims 1-4, characterized in that, The temperature of the seed culture is 28 - 32 °C; Preferably, the time of the seed culture is 24 - 72 h.

6. The fermentation method for increasing the fermentation yield of monensin according to any one of claims 1-5, characterized in that, The seed culture is carried out in a shaker at 100 - 400 rpm.

7. The fermentation method for increasing the fermentation yield of monensin according to any one of claims 1-6, characterized in that, The liquid loading amount of the fermentation medium is 1 / 6 - 1 / 3 of the volume of the culture container.

8. The fermentation method for increasing the fermentation yield of monensin according to any one of claims 1-7, characterized in that, The liquid loading amount of the seed medium is 1 / 12 - 1 / 8 of the volume of the culture container.

9. The fermentation method for increasing the fermentation yield of monensin according to any one of claims 1-8, characterized in that, The Streptomyces cinnamonensis is first subjected to solid culture before being inoculated into the seed medium: coating the strain on a solid medium and culturing it at 28 - 32 °C for 24 - 72 h.

10. Use of the fermentation method for increasing the fermentation yield of monensin according to any one of claims 1-9 in the preparation of monensin.

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