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Method for producing erythrocin by virtue of fermentation

A technology of erythromycin and fermentation culture, applied in the field of fermentative production of erythromycin, can solve the problems of no disclosure of strains, no discussion on the use of cottonseed oil, etc., and achieves low cost, is conducive to large-scale production, and improves content. Effect

Active Publication Date: 2015-03-18
鲁南新时代生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chinese invention patent application CN102732383A discloses a modified vegetable oil and its application in the fermentation process of erythromycin. The technical proposal discloses that the effect of modified cottonseed oil is better than that of natural soybean oil, but the technical proposal does not disclose the bacteria used It is well known that the impact of strains on fermentation technology is extremely important
The use of unmodified cottonseed oil is not addressed

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Slant culture: Prepare medium according to the following ratio: starch 10g / L, corn steep liquor 10g / L, sodium chloride 2g / L, ammonium sulfate 3g / L, calcium carbonate 3g / L, agar 22g / L, pH 7.0. Prepare 300ml of medium, heat the agar to melt, stir evenly, put it into 10 test tubes, sterilize at 121°C for 30min, and make it into a slope for later use. Dissolve the lyophilized red Saccharopolyspora erythraea (Saccharopolyspora erythraea) CPCC240502 strain in 5ml of sterile water, take 4ml and evenly inoculate it on 10 inclined surfaces, and cultivate it for 10 days at a temperature of 33°C and a humidity of 40%. Produces large numbers of spores. Add sterile water to make a spore suspension.

[0028] Shake flask seed solution preparation: Prepare medium according to the following ratio: sucrose 40g / L, ammonium sulfate 4g / L, corn steep liquor 15g / L, potassium dihydrogen phosphate 0.1g / L, magnesium sulfate 0.2g / L, calcium carbonate 7g / L L, refined cottonseed oil 0.05g / 50ml, p...

Embodiment 2

[0032] Slant culture: Prepare medium according to the following ratio: starch 12g / L, corn steep liquor 12g / L, sodium chloride 3g / L, ammonium sulfate 2g / L, calcium carbonate 4g / L, agar 25g / L, pH 7.5. Prepare 300ml of medium, heat the agar to melt, stir evenly, put it into 10 test tubes, sterilize at 121°C for 30min, and make it into a slope for later use. Dissolve freeze-dried red Saccharopolyspora (Saccharopolyspora erythraea) CPCC240502 in 5ml of sterile water, take 4ml and inoculate evenly on 10 slant surfaces, cultivate for 11 days at a temperature of 32°C and a humidity of 30%, and produce a large number of spores . Add sterile water to make a spore suspension.

[0033] Shake flask seed solution preparation: Prepare medium according to the following ratio: sucrose 50g / L, ammonium sulfate 6g / L, corn steep liquor 10g / L, potassium dihydrogen phosphate 0.15g / L, magnesium sulfate 0.1g / L, calcium carbonate 8g / L L, refined cottonseed oil 0.05g / 50ml, pH 6.0. Prepare 200ml of cu...

Embodiment 3

[0036] Embodiment 3 (comparative example)

[0037] Use soybean oil instead of refined cottonseed oil for the culture medium used in the shake flask seed liquid culture process, the seed tank medium used in the seed tank culture process, and the fermenter basal medium and feed in the fermenter culture process, and the others are implemented in the same way Example 2, the content of erythromycin was 3860U / ml when the tank was placed, and the content of erythromycin A was 67.9%.

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Abstract

The invention belongs to the field of biological fermentation and production and provides a method for producing erythrocin by virtue of fermentation. By taking saccharopolyspora erythraea as a starting germ, the method mainly comprises four steps of preparation of slant strains, preparation of shake-flask seed liquid, culture of seed liquid and process control of a fermentation tank. According to the method, soybean oil is replaced by refined cottonseed oil in a seed tank culture medium, a fermentation tank culture medium and the material supplementing process of a fermentation process, so that the fermentation cost is lowered; the fermentation valence and the content of erythrocin A are greatly increased based on an optimized fermentation process. The method is suitable for large-scale industrial production.

Description

technical field [0001] The invention belongs to the field of biological fermentation, and in particular relates to a method for producing erythromycin by fermentation. Background technique [0002] Erythromycin (Er) is a secondary metabolite produced by Saccharopolyspora erythraea and belongs to the macrolide antibiotics. Erythromycin has a strong antibacterial effect on aerobic Gram-positive cocci and bacilli, but is ineffective against most aerobic Gram-negative bacilli. However, other Gram-negative bacteria such as Neisseria gonorrhoeae, Haemophilus influenzae, Neisseria meningitidis, Pasteurella multocida, Treponema pallidum, Bordetella pertussis, Campylobacter jejuni and Legionella pneumophila, Mycoplasma pneumoniae, Chlamydia trachomatis has inhibitory effect. [0003] In recent years, due to the drug resistance of erythromycin itself and some other side effects, its own use has been affected, but the second-generation erythromycin derivatives developed later mostly ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/62C12N1/20C12R1/04
Inventor 赵志全谭丽华隋华芹孟祥学
Owner 鲁南新时代生物技术有限公司
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