Method for producing enramycin by using microbial fermentation

A technology of microbial fermentation and enramycin, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of unstable fermentation results, cumbersome seed production process, and high probability of bacterial infection, so as to save The effect of repeating the seeding steps, good repeatability, and improved fermentation results

Active Publication Date: 2012-09-05
山东胜利生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the production of enramycin usually adopts the traditional slant culture, the inoculation ring is dug to inoculate the shake flask, and the shake flask is inoculated with the seed tank. The seed production process is cumbersome, the probability of bacterial infection is high, the fermentation result is unstable, and the repeatability is poor

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] The selected SDSL1205 strain was used for fermentation.

[0082] (1) Preparation of frozen spore liquid:

[0083] a. Inoculate the bacterial strains on multiple plate mediums and culture them at 27°C for 7 days, the resulting spores are gray, round, with small protrusions in the middle;

[0084] b. Scrape the spores cultivated on each flat plate into 20wt% glycerin aqueous solution respectively, break up and mix with glass beads to make spore liquid, and the spore liquid is subpackaged in 25 cryopreservation tubes, and 1ml of spore liquid is adorned in each tube. Cryopreservation tubes were cryopreserved in a liquid nitrogen environment; the spore concentration was 10 8 -10 9 pcs / ml or so;

[0085] (2) Seed culture: inoculate the frozen spore liquid into the seed medium, inoculate 1.5 milliliters of the frozen spore liquid per 30 liters of culture medium, at 28.5 ° C, the ventilation rate is 1.0 vvm, and the aseptic culture is carried out for 50 h under the rotation ...

Embodiment 2

[0098] (1) Preparation of frozen spore liquid:

[0099] a. Inoculate the SDSL1205 strain on multiple plate media and culture at 28°C for 7 days;

[0100] b. Scrape the spores cultivated on each flat plate into 20wt% glycerol aqueous solution respectively, break up and mix with glass beads to make spore liquid, and the spore liquid is subpackaged in 20 cryopreservation tubes, and 1ml of spore liquid is adorned in each tube. Cryopreservation tubes were cryopreserved in a liquid nitrogen environment; the spore concentration was 10 8 -10 9 pcs / ml or so;

[0101] (2) Seed cultivation: inoculate the frozen spore liquid into the seed medium, inoculate 1 milliliter of the frozen spore liquid per 30 liters of culture medium, at 28° C., with an air flow of 1.1 vvm, and aseptically cultivate for 50 h under a rotating speed of 250 rpm to obtain seeds liquid; the solids in the seed liquid account for 20-30% by volume of the total seed liquid;

[0102] (3) Fermentation culture: inoculat...

Embodiment 3

[0111] (1) Preparation of frozen spore liquid:

[0112] a. Inoculate the SDSL1205 strain on multiple plate culture media and culture at 28°C for 6 days;

[0113] b. Scrape the spores cultivated on each flat plate into 20wt% glycerol aqueous solution respectively, break up and mix with glass beads to make spore liquid, and the spore liquid is subpackaged in 20 cryopreservation tubes, and 1ml of spore liquid is adorned in each tube. Cryopreservation tubes were cryopreserved in a liquid nitrogen environment; the spore concentration was 10 8 -10 9 pcs / ml or so;

[0114] (2) Seed cultivation: inoculate the frozen spore liquid into the seed medium, inoculate 1 milliliter of the frozen spore liquid per 30 liters of culture medium, at 28° C., with an air flow of 1.1 vvm, and aseptically cultivate for 50 h under a rotating speed of 250 rpm to obtain seeds liquid; the solids in the seed liquid account for 20-30% by volume of the total seed liquid;

[0115] (3) Fermentation culture: ...

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Abstract

The invention discloses a method for producing enramycin by using microbial fermentation. The method comprises the following steps of: preparing spore liquor of streptomyces fungicidicus SDSL1205 CGMCC No. 3933, and preserving the spore liquor under the environment of liquid nitrogen; inoculating the frozen and preserved spore liquor into a seed culture medium to culture seeds so as to obtain seed liquor; inoculating the seed liquor into a fermentation medium for fermenting and culturing according to the inoculum size of between 4 and 6 weight percent; and further processing fermented fermentation liquor to obtain enramycin crystals. The method has the advantages of omitting complicated seed making process, reducing microbiological contamination rate, along with simpleness and practicability, easy control of inoculum size, stable fermentation result and high repeatability. Compared with the conventional process, the method improves the fermentation result to some extent; and the maximum yield of the enramycin is over 8,500 mu g / ml (HPLC).

Description

technical field [0001] The invention relates to a method for producing enramycin by microbial fermentation, which belongs to the technical field of biological fermentation. Background technique [0002] Enramycin, also known as anlemycin, is a polypeptide antibiotic produced by fermentation of the actinomyces Streptomyces fungicidious NO.B5477 isolated from the soil. Enramycin was named Enduracidin because of its strong bactericidal effect, and then the World Health Organization (WHO) determined its general name as Enramycin. Enramycin is an organic base, and its hydrochloride is a white crystalline powder with a molecular weight of about 2500 and a melting point of 238-245°C. This product is easily soluble in dilute hydrochloric acid and dimethyl sulfoxide, soluble in methanol and hydrous ethanol, insoluble in ethanol and acetone, insoluble in benzene, chloroform, etc. The chemical structure of enramycin can be determined by partial hydrolysis of peptide bonds. It has be...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/04C12N1/20C12R1/465
Inventor 胡江林王永星景巍葛庆燕
Owner 山东胜利生物工程有限公司
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