Application of ethyl linoleate in improving nystatin yield and a fermentation method for high-yield nystatin

Abstract

The application belongs to the technical field of fermentation engineering, and relates to application of ethyl linoleate in improving nystatin yield and a fermentation method for high-yield nystatin. The application provides the application of ethyl linoleate in improving the nystatin yield. By using ethyl linoleate as a feed, the yield of nystatin in the process of fermentative production of nystatin can be improved.

Description

Technical Field

[0001] This invention belongs to the field of fermentation engineering technology, specifically relating to the application of ethyl linoleate in increasing nystatin yield and a fermentation method for high-yield nystatin. Background Technology

[0002] Nystatin, also known as fungicidin or mycosta-tin, is a 38-membered polyene macrolide antibiotic with the molecular formula C3. 47 H 75 NO 17 The compound has a hydroxyl group at C10, a carboxyl group at C16, and a trehalose-amine group at C19. Nystatin is usually found in mycelium. Its pure form is a pale yellow crystal that is easily decomposed by light and heat. It is usually stored frozen in crystalline form. It is hygroscopic, insoluble in acetone and chloroform, very slightly soluble in water, slightly soluble in lower alcohols, and soluble in N,N-dimethylformamide. It is unstable at high or low pH and its solution state is also unstable. It has ultraviolet absorption at wavelengths of 291 nm, 305 nm, and 319 nm.

[0003] The biosynthesis of nystatin follows a typical type I polyketide synthetase (PKS) pathway. Modification of the synthesis process using modern biotechnology can yield structural analogs with different properties. However, the low yield remains a challenge in the biosynthesis of nystatin. Summary of the Invention

[0004] The purpose of this invention is to provide the application of ethyl linoleate in increasing nystatin yield and a fermentation method for high-yield nystatin production. By using ethyl linoleate as a feedstock, the yield of nystatin during fermentation production can be increased.

[0005] This invention provides the application of ethyl linoleate in increasing nystatin yield.

[0006] Preferably, in the application, ethyl linoleate is used as a feedstock to increase the yield of nystatin by fermentation using the nystatin-producing strain.

[0007] Preferably, the nystatin-producing strain is Streptomyces noursei ATCC11455.

[0008] This invention also provides a fermentation method for increasing nystatin yield, comprising the following steps:

[0009] Feed-fed fermentation of the nystatin-producing strain was carried out using ethyl linoleate.

[0010] Preferably, the nystatin-producing strain is Streptomyces noursei ATCC11455.

[0011] Preferably, during the fed-batch fermentation process, the concentration of ethyl linoleate is controlled at 10–20 g / L.

[0012] Preferably, feeding is carried out when dissolved oxygen levels rise, and during the feeding fermentation process, dissolved oxygen is maintained above 50%.

[0013] Preferably, the pH value before feeding is 6.2 to 6.9.

[0014] Preferably, the pH value is 6.9 to 7.3 during the fed-batch fermentation process.

[0015] Preferably, the pH value is adjusted using ammonia.

[0016] This invention provides the application of ethyl linoleate in increasing nystatin production. This invention discovers that by using ethyl linoleate as a feedstock, nystatin can be produced efficiently, increasing the yield of a 100L automated control tank from 200,000 IU / g to 600,000 IU / g. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0018] Figure 1 The image shows the HPLC detection results of nystatin content provided in Example 1 of this invention.

[0019] Figure 2 This is a graph showing the HPLC detection results of nystatin content provided in Example 2 of the present invention;

[0020] Figure 3 This is a graph showing the HPLC detection results of nystatin content provided in Example 3 of the present invention;

[0021] Figure 4 This is a graph showing the HPLC detection results of nystatin content provided in Example 4 of the present invention;

[0022] Figure 5 This is a graph showing the HPLC detection results of nystatin content provided in Example 5 of the present invention;

[0023] Figure 6 This is a graph showing the HPLC detection results of nystatin content provided in Example 6 of the present invention;

[0024] Figure 7 The HPLC liquid phase detection result of nystatin content provided in Example 7 of the present invention (138h);

[0025] Figure 8 The HPLC liquid phase detection result of nystatin content provided in Example 7 of the present invention (162h). Detailed Implementation

[0026] This invention provides the application of ethyl linoleate in increasing nystatin yield. In the biosynthesis of nystatin, i.e., fermentation production, this invention discovers that using ethyl linoleate as a feedstock can increase nystatin yield. Preferably, in this invention, ethyl linoleate is used as a feedstock, and nystatin-producing strains are used to increase nystatin yield through fermentation. In this invention, the preferred nystatin-producing strain is *Streptomyces noursei* ATCC 11455. This invention uses ethyl linoleate as a precursor carbon source for the fermentation production of nystatin, which is more easily hydrolyzed and metabolized to produce short-chain fatty acids. Simultaneously, it produces more acetyl-CoA, enhancing the activity of acyltransferases during metabolism, thereby strengthening the PKs pathway (polyketide synthase pathway). Furthermore, ethyl linoleate itself has a certain defoaming ability; after its addition, foam during fermentation is effectively controlled, improving the loading coefficient and increasing the yield per batch.

[0027] This invention also provides a fermentation method for increasing nystatin yield, comprising the following steps:

[0028] Feed-fed fermentation of the nystatin-producing strain was carried out using ethyl linoleate.

[0029] In this invention, the nystatin-producing strain is Streptomyces noursei ATCC11455.

[0030] In this invention, prior to the fed-batch fermentation, a seed tank fermentation and a fermenter fermentation process are preferably performed. Prior to the seed tank fermentation, seed bottle culture is preferably performed. The preferred conditions for seed bottle culture are 30°C, 250 rpm, and culture until OD (dose retardation). 600=0.5. In this invention, the culture medium added to the seed bottle is preferably TSB medium. In this invention, the inoculation amount of the seed liquid obtained from the seed bottle culture in the seed tank is preferably 1% to 2%. In this invention, the seed culture medium used for fermentation culture in the seed tank preferably includes 4 to 8 g / L soybean flour, 7 to 12 g / L dry yeast, 6 to 12 g / L peptone, 1 to 2 g / L light calcium carbonate, 1 to 5 g / L KH2PO4, and 20 to 30 g / L glucose monohydrate, with a natural pH. In this invention, the defoamer is preferably a polyether defoamer, more preferably purchased from Yantai Hengxin Chemical Technology Co., Ltd., product number HIX-286. In this invention, the culture medium for fermentation preferably comprises 20-30 g / L soybean flour, 40-60 g / L dry yeast, 6-12 g / L corn gluten flour, 1-2 g / L light calcium carbonate, 1-5 g / L KH₂PO₄, and 20-30 g / L glucose monohydrate, with a pH of 6.6-7.0. In this invention, the preferred temperature for both the seed tank fermentation and the fermentation tank fermentation is 30°C. In this invention, the preferred rotation speed for both the seed tank fermentation and the fermentation tank fermentation is 200 rpm.

[0031] In this invention, during the fed-batch fermentation process, the concentration of ethyl linoleate is preferably controlled at 10–20 g / L, more preferably at 15 g / L. In this invention, feeding is preferably carried out when dissolved oxygen levels rise, and during the fed-batch fermentation process, dissolved oxygen is preferably maintained above 50%. Before feeding, ethyl linoleate is preferably sterilized. In this invention, the sterilization conditions are preferably 121°C for 30 min. In this invention, the concentration of ethyl linoleate is preferably detected using conventional methods. For example, 10 ml of fermentation broth is centrifuged at 3500 rpm for 10 min, the volume of ethyl linoleate in the supernatant is recorded, and then the concentration of ethyl linoleate in the fermentation broth is calculated.

[0032] In this invention, the pH value before feeding is preferably 6.2 to 6.9. In this invention, during the fed-batch fermentation process, the pH value is preferably 6.9 to 7.3. In this invention, the pH value is preferably adjusted using ammonia.

[0033] To further illustrate the present invention, the application of ethyl linoleate provided by the present invention in increasing nystatin yield and a fermentation method for high-yield nystatin are described in detail below with reference to the accompanying drawings and embodiments. However, these descriptions should not be construed as limiting the scope of protection of the present invention.

[0034] Example 1

[0035] 100L automated fermentation tank

[0036] Seed bottle inoculation: Take two working cell bank glycerol tubes (Streptomyces noursei ATCC 11455) from the ultra-low temperature freezer, and pipette 0.5 mL of seed culture into seed bottles. Inoculate 6 bottles and place them in a culture room on a shaker at 30°C and 250 rpm for 6 hours. After incubation, randomly select one bottle of seed culture for pH and OD analysis. 600 The detection of process control indicators such as glucose, and staining microscopy, were used to determine the seed attainment index (OD). 600 =0.5), after sterilization, the bottles can be combined and put into the can.

[0037] Seed culture in a clean bench: Combine the cultured seeds into two bottles, about 140 mL each (1.5% inoculum). Seeds must be inoculated with broth slant medium for aseptic monitoring before being placed into the culture vessel.

[0038] Seed tank: Before merging the bottles into the tank, dissolve all raw materials except for the defoamer (purchased from Yantai Hengxin Chemical Technology Co., Ltd., product number HIX-286) and light calcium carbonate in water and mix thoroughly. Adjust the pH to 6.5 with ammonia solution, add CaCO3 and the defoamer. Sterilize at 121℃ for 30 minutes.

[0039] Seed culture medium formula: 6 g / L soybean flour, 10 g / L dry yeast, 9 g / L peptone, 1.5 g / L light calcium carbonate, 3 g / L KH2PO4, 25 g / L glucose monohydrate, natural pH.

[0040] Fermentation tank: Dissolve all raw materials except defoamer and light calcium carbonate in water, adjust the pH to 6.5 with ammonia solution, add CaCO3 and defoamer, and sterilize at 121℃ for 30 minutes. When the pH of the seed tank drops to about 0.8, transfer to the fermentation tank, with an inoculum size of 30%.

[0041] Fermentation medium formula: 25 g / L soybean flour, 50 g / L dry yeast, 9 g / L corn gluten flour, 1.5 g / L light calcium carbonate, 3 g / L KH2PO4, 25 g / L glucose monohydrate, pH 6.5.

[0042] Process control: Dissolved oxygen is maintained above 50%, ammonia water pH is controlled at 7.1, ethyl linoleate level is controlled at 15g / L, and the discharge potency is 594975IU / g.

[0043] Example 2

[0044] 100L automated fermentation tank

[0045] Seed bottle inoculation: Take two working cell bank glycerol tubes (Streptomyces noursei ATCC 11455) from the ultra-low temperature freezer, and pipette 0.5 mL of seed culture into seed bottles. Inoculate 6 bottles and place them in a culture room on a shaker at 30°C and 250 rpm for 6 hours. After incubation, randomly select one bottle of seed culture for pH and OD analysis. 600 The detection of process control indicators such as glucose, and staining microscopy, were used to determine the seed attainment index (OD). 600 =0.5), after sterilization, the bottles can be combined and put into the can.

[0046] Seed culture in a clean bench: Combine the cultured seeds into two bottles, about 140 mL each (1.5% inoculum). Seeds must be inoculated with broth slant medium for aseptic monitoring before being placed into the culture vessel.

[0047] Seed tank: Before merging the bottles into the tank, dissolve all raw materials except for the defoamer (as above) and light calcium carbonate in water and mix thoroughly. Adjust the pH to 6.5 with ammonia solution, then add CaCO3 and the defoamer. Sterilize at 121℃ for 30 minutes.

[0048] Seed culture medium formula: 6 g / L soybean flour, 10 g / L dry yeast, 9 g / L peptone, 1.5 g / L light calcium carbonate, 3 g / L KH2PO4, 25 g / L glucose monohydrate, natural pH.

[0049] Fermentation tank: Dissolve all raw materials except defoamer and light calcium carbonate in water, adjust the pH to 6.6 with ammonia solution, add CaCO3 and defoamer, and sterilize at 121℃ for 30 minutes. When the pH of the seed tank drops to about 0.8, transfer to the fermentation tank, with an inoculum size of 30%.

[0050] Fermentation medium formula: 25 g / L soybean flour, 50 g / L dry yeast, 9 g / L corn gluten flour, 1.5 g / L light calcium carbonate, 3 g / L KH2PO4, 25 g / L glucose monohydrate, pH 6.6.

[0051] Process control: Dissolved oxygen is maintained above 50%, ammonia water pH is controlled at 7.1, ethyl linoleate level is controlled at 15g / L, and the discharge potency is 613227IU / g.

[0052] Example 3

[0053] 100L automated fermentation tank

[0054] Seed bottle inoculation: Take two working cell bank glycerol tubes (Streptomyces noursei ATCC 11455) from the ultra-low temperature freezer, and pipette 0.5 mL of seed culture into seed bottles. Inoculate 6 bottles and place them in a culture room on a shaker at 30°C and 250 rpm for 6 hours. After incubation, randomly select one bottle of seed culture for pH and OD analysis. 600 The detection of process control indicators such as glucose, and staining microscopy, were used to determine the seed attainment index (OD). 600 =0.5), after sterilization, the bottles can be combined and put into the can.

[0055] Seed culture in a clean bench: Combine the cultured seeds into two bottles, about 140 mL each (1.5% inoculum). Seeds must be inoculated with broth slant medium for aseptic monitoring before being placed into the culture vessel.

[0056] Seed tank: Before merging the bottles into the tank, dissolve all raw materials except for the defoamer (as above) and light calcium carbonate in water and mix thoroughly. Adjust the pH to 6.5 with ammonia solution, then add CaCO3 and the defoamer. Sterilize at 121℃ for 30 minutes.

[0057] Seed culture medium formula: 6 g / L soybean flour, 10 g / L dry yeast, 9 g / L peptone, 1.5 g / L light calcium carbonate, 3 g / L KH2PO4, 25 g / L glucose monohydrate, natural pH.

[0058] Fermentation tank: Dissolve all raw materials except defoamer and light calcium carbonate in water, adjust the pH to 6.4 with ammonia solution, add CaCO3 and defoamer, and sterilize at 121℃ for 30 minutes. When the pH of the seed tank drops to about 0.8, transfer to the fermentation tank, with an inoculum size of 30%.

[0059] Fermentation medium formula: 25 g / L soybean flour, 50 g / L dry yeast, 9 g / L corn gluten flour, 1.5 g / L light calcium carbonate, 3 g / L KH2PO4, 25 g / L glucose monohydrate, pH 6.4.

[0060] Process control: Dissolved oxygen is maintained above 50%, ammonia water pH is controlled at 7.1, ethyl linoleate level is controlled at 15g / L, and the discharge potency is 623781IU / g.

[0061] Example 4

[0062] 100L automated fermentation tank

[0063] Seed bottle inoculation: Take two working cell bank glycerol tubes (Streptomyces noursei ATCC 11455) from the ultra-low temperature freezer, and pipette 0.5 mL of seed culture into seed bottles. Inoculate 6 bottles and place them in a culture room on a shaker at 30°C and 250 rpm for 6 hours. After incubation, randomly select one bottle of seed culture for pH and OD analysis. 600 The detection of process control indicators such as glucose, and staining microscopy, were used to determine the seed attainment index (OD). 600 =0.5), after sterilization, the bottles can be combined and put into the can.

[0064] Seed culture in a clean bench: Combine the cultured seeds into two bottles, about 140 mL each (1.5% inoculum). Seeds must be inoculated with broth slant medium for aseptic monitoring before being placed into the culture vessel.

[0065] Seed tank: Before merging the bottles into the tank, dissolve all raw materials except for the defoamer (as above) and light calcium carbonate in water and mix thoroughly. Adjust the pH to 6.5 with ammonia solution, then add CaCO3 and the defoamer. Sterilize at 121℃ for 30 minutes.

[0066] Seed culture medium formula: 6 g / L soybean flour, 10 g / L dry yeast, 9 g / L peptone, 1.5 g / L light calcium carbonate, 3 g / L KH2PO4, 25 g / L glucose monohydrate, natural pH.

[0067] Fermentation tank: Dissolve all raw materials except defoamer and light calcium carbonate in water, adjust the pH to 6.4 with ammonia solution, add CaCO3 and defoamer, and sterilize at 121℃ for 30 minutes. When the pH of the seed tank drops to about 0.8, transfer to the fermentation tank, with an inoculum size of 30%.

[0068] Fermentation medium formula: 25 g / L soybean flour, 50 g / L dry yeast, 9 g / L corn gluten flour, 1.5 g / L light calcium carbonate, 3 g / L KH2PO4, 25 g / L glucose monohydrate, pH 6.4.

[0069] Process control: Dissolved oxygen is maintained above 50%, ammonia water pH is controlled at 7.1, glucose level is controlled at 4g / L, and the discharge potency is 209640IU / g.

[0070] Example 5

[0071] 100L automated fermentation tank

[0072] Seed bottle inoculation: Take two working cell bank glycerol tubes (Streptomyces noursei ATCC 11455) from the ultra-low temperature freezer, and pipette 0.5 mL of seed culture into seed bottles. Inoculate 6 bottles and place them in a culture room on a shaker at 30°C and 250 rpm for 6 hours. After incubation, randomly select one bottle of seed culture for pH and OD analysis. 600 The detection of process control indicators such as glucose, and staining microscopy, were used to determine the seed attainment index (OD). 600 =0.5), after sterilization, the bottles can be combined and put into the can.

[0073] Seed culture in a clean bench: Combine the cultured seeds into two bottles, about 140 mL each (1.5% inoculum). Seeds must be inoculated with broth slant medium for aseptic monitoring before being placed into the culture vessel.

[0074] Seed tank: Before merging the bottles into the tank, dissolve all raw materials except for the defoamer (as above) and light calcium carbonate in water and mix thoroughly. Adjust the pH to 6.5 with ammonia solution, then add CaCO3 and the defoamer. Sterilize at 121℃ for 30 minutes.

[0075] Seed culture medium formula: 6 g / L soybean flour, 10 g / L dry yeast, 9 g / L peptone, 1.5 g / L light calcium carbonate, 3 g / L KH2PO4, 25 g / L glucose monohydrate, natural pH.

[0076] Fermentation tank: Dissolve all raw materials except defoamer and light calcium carbonate in water, adjust the pH to 6.6 with ammonia solution, add CaCO3 and defoamer, and sterilize at 121℃ for 30 minutes. When the pH of the seed tank drops to about 0.8, transfer to the fermentation tank, with an inoculum size of 30%.

[0077] Fermentation medium formula: 25 g / L soybean flour, 50 g / L dry yeast, 9 g / L corn gluten flour, 1.5 g / L light calcium carbonate, 3 g / L KH2PO4, 25 g / L glucose monohydrate, pH 6.6.

[0078] Process control: Dissolved oxygen is maintained above 50%, ammonia water pH is controlled at 7.1, glucose level is controlled at 4g / L, and the discharge potency is 210984IU / g.

[0079] Example 6

[0080] 100L automated fermentation tank

[0081] Seed bottle inoculation: Take two working cell bank glycerol tubes (Streptomyces noursei ATCC 11455) from the ultra-low temperature freezer, and pipette 0.5 mL of seed culture into seed bottles. Inoculate 6 bottles and place them in a culture room on a shaker at 30°C and 250 rpm for 6 hours. After incubation, randomly select one bottle of seed culture for pH and OD analysis. 600 The detection of process control indicators such as glucose, and staining microscopy, were used to determine the seed attainment index (OD). 600 =0.5), after sterilization, the bottles can be combined and put into the can.

[0082] Seed culture in a clean bench: Combine the cultured seeds into two bottles, about 140 mL each (1.5% inoculum). Seeds must be inoculated with broth slant medium for aseptic monitoring before being placed into the culture vessel.

[0083] Seed tank: Before merging the bottles into the tank, dissolve all raw materials except for the defoamer (as above) and light calcium carbonate in water and mix thoroughly. Adjust the pH to 6.5 with ammonia solution, then add CaCO3 and the defoamer. Sterilize at 121℃ for 30 minutes.

[0084] Seed culture medium formula: 6 g / L soybean flour, 10 g / L dry yeast, 9 g / L peptone, 1.5 g / L light calcium carbonate, 3 g / L KH2PO4, 25 g / L glucose monohydrate, natural pH.

[0085] Fermentation tank: Dissolve all raw materials except defoamer and light calcium carbonate in water, adjust the pH to 6.5 with ammonia solution, add CaCO3 and defoamer, and sterilize at 121℃ for 30 minutes. When the pH of the seed tank drops to about 0.8, transfer to the fermentation tank, with an inoculum size of 30%.

[0086] Fermentation medium formula: 25 g / L soybean flour, 50 g / L dry yeast, 9 g / L corn gluten flour, 1.5 g / L light calcium carbonate, 3 g / L KH2PO4, 25 g / L glucose monohydrate, pH 6.5.

[0087] Process control: Dissolved oxygen is maintained above 50%, ammonia water pH is controlled at 7.1, glucose level is controlled at 4g / L, and the discharge potency is 209745IU / g.

[0088] Example 7

[0089] 100L automated fermentation tank

[0090] Seed bottle inoculation: Take two working cell bank glycerol tubes (Streptomyces noursei ATCC 11455) from the ultra-low temperature freezer, and pipette 0.5 mL of seed culture into seed bottles. Inoculate 6 bottles and place them in a culture room on a shaker at 30°C and 250 rpm for 6 hours. After incubation, randomly select one bottle of seed culture for pH and OD analysis. 600 The detection of process control indicators such as glucose, and staining microscopy, were used to determine the seed attainment index (OD). 600 =0.5), after sterilization, the bottles can be combined and put into the can.

[0091] Seed culture in a clean bench: Combine the cultured seeds into two bottles, about 140 mL each (1.5% inoculum). Seeds must be inoculated with broth slant medium for aseptic monitoring before being placed into the culture vessel.

[0092] Seed tank: Before merging the bottles into the tank, dissolve all raw materials except for the defoamer (as above) and light calcium carbonate in water and mix thoroughly. Adjust the pH to 6.5 with ammonia solution, then add CaCO3 and the defoamer. Sterilize at 121℃ for 30 minutes.

[0093] Seed culture medium formula: 6 g / L soybean flour, 10 g / L dry yeast, 9 g / L peptone, 1.5 g / L light calcium carbonate, 3 g / L KH2PO4, 25 g / L glucose monohydrate, natural pH.

[0094] Fermentation tank: Dissolve all raw materials except defoamer and light calcium carbonate in water, adjust the pH to 6.5 with ammonia solution, add CaCO3 and defoamer, and sterilize at 121℃ for 30 minutes. When the pH of the seed tank drops to about 0.8, transfer to the fermentation tank, with an inoculum size of 30%.

[0095] Fermentation medium formula: 25 g / L soybean flour, 50 g / L dry yeast, 9 g / L corn gluten flour, 1.5 g / L light calcium carbonate, 3 g / L KH2PO4, 25 g / L glucose monohydrate, pH 6.5.

[0096] Process control: Dissolved oxygen was maintained above 50%, ammonia water was controlled at pH 7.1, and glucose level was controlled at 4g / L. In this example, the fermentation time was extended. After 138h of cultivation, the potency was 208694IU / g. After that, the potency increase became smaller. After 162h, the product was discharged from the tank, and the potency was 219207IU / g.

[0097] The results are shown in Table 1:

[0098] Table 1. Fermentation results of Examples 1-6

[0099] Example 1 310 70 594975 Example 2 331 69 613227 Example 3 311 69 623781 Example 4 143 61 209640 Example 5 143 60 210984 Example 6 142 60 209745 Example 7 162 58 219207

[0100] The fermentation products obtained in Examples 1 to 7 were analyzed by HPLC, and the results are as follows: Figures 1 to 8 .according to Figures 1 to 8 It is evident that extending the culture time using glucose does not effectively improve potency. Replacing glucose with ethyl linoleate as the feedstock can effectively extend the fermentation time. Furthermore, ethyl linoleate has an antifoaming effect, which can increase the loading coefficient and significantly improve the output of the automated control tank.

[0101] Compared with Examples 4-6, Examples 1-3 have the advantage that the present invention uses ethyl linoleate for feeding. While serving as a carbon source, ethyl linoleate, being a strongly hydrophobic and weakly hydrophilic linear molecule, reduces the local surface tension of the foam when added to the fermentation broth, leading to foam rupture and thus defoaming.

[0102] Compared with Example 7, Examples 1-3 have the advantage that the present invention uses ethyl linoleate for feeding, which effectively prolongs the fermentation time and greatly improves the fermentation level.

[0103] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Application of ethyl linoleate in increasing nystatin yield.

2. The application according to claim 1, characterized in that, In this application, ethyl linoleate is used as a feedstock, and the nystatin-producing strain is used to increase the yield of nystatin through fermentation.

3. The application according to claim 2, characterized in that, The nystatin-producing strain is Streptomyces noursei ATCC 11455.

4. A fermentation method for increasing nystatin yield, characterized in that, Includes the following steps: Feed-fed fermentation of the nystatin-producing strain was carried out using ethyl linoleate.

5. The fermentation method according to claim 4, characterized in that, The nystatin-producing strain is Streptomyces noursei ATCC 11455.

6. The fermentation method according to claim 4, characterized in that, During the fed-batch fermentation process, the concentration of ethyl linoleate is controlled at 10–20 g / L.

7. The fermentation method according to claim 4, characterized in that, Feeding is carried out when dissolved oxygen levels rise, and dissolved oxygen is maintained above 50% during the fed fermentation process.

8. The fermentation method according to claim 4, characterized in that, The pH value before feeding was 6.2 to 6.

9.

9. The fermentation method according to claim 4, characterized in that, During the fed-batch fermentation process, the pH value is 6.9–7.

3.

10. The fermentation method according to claim 8 or 9, characterized in that, The pH value is adjusted using ammonia.

Images