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Method for producing enramycin by using microbial fermentation

A technology of microbial fermentation and enramycin, which is applied in the directions 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 contamination, so as to save the Repeated seeding steps, good reproducibility, improved fermentation results

Active Publication Date: 2010-12-01
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] Fermentation culture was carried out using the selected strain SDSL1205.

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

[0083] a. The strains were inoculated on multiple plate media, and cultured at 27°C for 7 days, the obtained spores were gray, round, with small protrusions in the middle;

[0084] b. Scrape the spores cultured on each plate into a 20wt% glycerol aqueous solution, and mix them with glass beads to make spore liquid. Cryovials are cryopreserved in liquid nitrogen; spore concentration is 10 8 -10 9 pcs / ml or so;

[0085] (2) Seed culture: inoculate the cryopreserved spore solution into the seed medium, inoculate 1.5 ml of the cryopreserved spore solution per 30 liters of culture medium, and aseptically cultivate for 50h at 28.5°C, aeration volume of 1.0vvm, and rotation speed of 200rpm to obtain seeds. liquid; the solids in the seed liquid account for 20-30% by volume of the total amount of the seed liquid;

[0086] (3) Fermentation culture: th...

Embodiment 2

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

[0099] a. The SDSL1205 strain was inoculated onto multiple plate media and cultured at 28°C for 7 days;

[0100] b. Scrape the spores cultured on each plate into a 20wt% glycerol aqueous solution, and mix them with glass beads to make a spore liquid. Cryovials are cryopreserved in liquid nitrogen; spore concentration is 10 8 -10 9 pcs / ml or so;

[0101] (2) Seed culture: inoculate the cryopreserved spore solution into the seed medium, inoculate 1 ml of the cryopreserved spore solution per 30 liters of culture medium, and aseptically cultivate for 50 hours at 28° C., aeration volume of 1.1 vvm, and 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 amount of the seed liquid;

[0102] (3) Fermentation culture: the seed liquid is inoculated into the fermentation medium at 5% of the inoculation amount, and the fermentation culture is carried out for 300h...

Embodiment 3

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

[0112] a. The SDSL1205 strain was inoculated onto multiple plate media and cultured at 28°C for 6 days;

[0113] b. Scrape the spores cultured on each plate into a 20wt% glycerol aqueous solution, and mix them with glass beads to make a spore liquid. Cryovials are cryopreserved in liquid nitrogen; spore concentration is 10 8 -10 9 pcs / ml or so;

[0114] (2) Seed culture: inoculate the cryopreserved spore solution into the seed medium, inoculate 1 ml of the cryopreserved spore solution per 30 liters of culture medium, and aseptically cultivate for 50 hours at 28° C., aeration volume of 1.1 vvm, and 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 amount of the seed liquid;

[0115] (3) Fermentation culture: the seed liquid is inoculated into the fermentation medium at 5% of the inoculation amount, and the fermentation culture is carried out at 28° C...

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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, and belongs to the technical field of biological fermentation. Background technique [0002] Enramycin, also known as enramycin, is a polypeptide antibiotic produced by fermentation of actinomycetes Streptomyces fungicidious NO.B5477 isolated from soil. Enramycin was named Enduracidin because of its powerful bactericidal effect, and then the World Health Organization (WHO) determined its generic name as Enramycin. Enramycin is an organic base, and its hydrochloride salt 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, aqueous 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 been c...

Claims

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Application Information

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Patent Type & Authority Applications(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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