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Method for producing polypeptide enramycin with zymotechnics

A technology of polypeptide antibiotics and amycin, which is applied in the fields of agricultural biotechnology, industrial biotechnology, and fermentation engineering. It can solve the problems of complex process, low unit potency, and long fermentation time of anlamycin, and achieve strong antimicrobial properties. The effect of high activity, high unit potency and low production cost

Inactive Publication Date: 2008-08-20
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current amamycin process is complicated, the fermentation time is long, the production cost is high, and the unit potency is low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Streptomyces sp.NJWGY3665 strains were connected to 0.5% glucose, 0.5% beef extract, 0.5% peptone, 0.5% NaCl, 1.5% agar, 96.5% water, pH6.0 glucose nutrient medium, and carried out slant culture. Incubate at 28°C for 2 days. Take the slant bacteria, add 2-3ml sterile water (per 18×180mm test tube slant bacteria), gently wash the bacteria moss with an inoculation loop to make a spore suspension, take 0.5ml of the spore suspension and inoculate the containing 40ml Glucose 1.0%, Corn Steep Steep 3%, CaCO 3 1%, 0.5% yeast extract, 0.1% NaCl, 94.4% water, and cultured in a liquid seed medium of pH 7.0 for 3 days, the culture temperature was 28° C., and the rotation speed was 200 rpm. Then 5m1 of the seed solution was inserted into a 500ml shake flask with a liquid content of 100ml fermentation medium, and at 28°C, 180rpm, pH6.0, shaker fermentation culture was carried out for 5 days to obtain Streptomyces sp.NJWGY3665, using The method of centrifugation extracts and separ...

Embodiment 2

[0023] Streptomyces sp.NJWGY3665 strains were inoculated with glucose 0.5%, beef extract 0.8%, peptone 0.8%, K 2 HPO 4 0.1%, 1.8% agar, 96% water, pH 7.0 glucose nutrient medium, cultured on a slant, and cultured at 37°C for 3 days. Take the slant bacteria, add 2-3ml sterile water (per 18×180mm test tube slant bacteria), gently wash the bacteria lawn with an inoculation loop to make a spore suspension, take 0.5ml of the spore suspension and inoculate the spores The suspension was inoculated into 40ml of glucose 2.0%, corn steep liquor 2.0%, CaCO 3 1.5%, 0.5% yeast powder, 0.5% NaCl, 93.5% water, and cultured for 4 days in a liquid seed medium with pH 7.0, the culture temperature was 30° C., and the rotation speed was 200 rpm. Then 10ml of seed solution was inserted into a 500ml shake flask with a liquid volume of 100ml fermentation medium, at 32°C, 220rpm, shaking table fermentation and cultivation under the conditions of pH7.0 for 6 days to obtain Streptomyces sp.NJWGY366...

Embodiment 3

[0028] Use broth medium to connect Streptomyces sp.NJWGY3665 strains to glucose nutrient medium with 1% glucose, 1% beef extract, 1% peptone, 0.5% NaCl, 2% agar, 94.5% water, pH9.0 , cultured on a slant, and cultured at 32°C for 5 days. Take the slant bacteria, add 2-3ml sterile water (per 18×180mm test tube slant bacteria), gently wash the bacteria lawn with an inoculation loop to make a spore suspension, take 1ml of the spore suspension and inoculate to 50ml Glucose 3.0%, Corn Steep Steep 1%, CaCO 3 2%, 0.5% yeast powder, 0.8% NaCl, 92.7% water, and cultured for 4 days in a liquid seed medium with a pH of 9.0, the culture temperature was 32° C., and the rotation speed was 200 rpm. Then inoculate 50ml of seed solution into a 2L fermenter with a liquid capacity of 1.5L fermentation medium. At 37°C, air flows in at a total flow rate of 1.0L / min, and stirs and mixes at 400rpm to maintain the dissolved oxygen concentration (DO ) is 60-100%. Use 20% (w / v) NaOH to maintain the ...

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Abstract

The invention relates to a method for producing polypeptide antibiotics enramycin by the fermentation method, which pertains to the fields of agricultural biotechnology, industrial biotechnology and the fermentation engineering. The method is as follows: (1) slant culture: a Streptomyces sp.NJWGY3665 bacterial strain is inoculated in a glucose nutrient culture medium to carry out the slant culture, the culture temperature is 28 to 37 DEG C and the culture time is 2 to 6 days; (2) seed culture: a spore which is cultured on a slant is produced into a single-spore suspension by using sterile water, which is also inoculated in a seed culture medium for culture, the temperature is 28 to 32 DEG C, the rotational speed is 200rpm and the culture lasts for 2 to 6 days; (3) fermentation culture: the seed liquid is inoculated in a fermentation culture medium for culture, the temperature is controlled at 28 to 37 DEG C, the pH is controlled at 6.0 to 9.0, the rotational speed is 180 to 220rpm, the fermentation lasts for 5 to 9 days, so as to obtain the Streptomyces sp., methanol or ethanol is used for the extraction and separation of mycelium through the method of centrifugal separation, then the resin method is adopted for carrying out refining, so as to obtain the peptide antibiotics enramycin.

Description

technical field [0001] The invention relates to a fermentation method for producing polypeptide antibiotic amellamycin, which belongs to the fields of agricultural biotechnology, industrial biotechnology and fermentation engineering. Background technique [0002] It has been nearly 60 years since antibiotics were added to feed to promote the growth and development of livestock and improve feed. Many countries have used antibiotic additives for a long time. However, with the widespread use of antibiotic growth promoters, a series of problems have gradually emerged, mainly drug residues and drug resistance. The long-term, extensive and indiscriminate use of antibiotic feed has brought great trouble and difficulty to the control and treatment of bacterial diseases, and also brought potential threats to human health. Although there are many controversies about whether antibiotics should be used in feed, practice has proved that rational use of antibiotic additives can promote t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/00C12R1/465
Inventor 胡永红杨文革
Owner NANJING UNIV OF TECH
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