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Method for producing amphotericin B through fermentation

A technology of streptomycin and fermentation broth, applied in the field of fermentation engineering, can solve the problems of unstable fermentation process and low fermentation level of amphotericin B, and achieve the effects of shortening production cycle, shortening fermentation time and ensuring consistency

Active Publication Date: 2016-04-27
NCPC NEW DRUG RES & DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to solve the problem of unstable fermentation process and low fermentation level in the preparation of amphotericin B by microbial fermentation in the prior art, and to provide a method of adding precursors in the fermentation process by optimizing the fermentation medium formula and fermentation conditions. Substance, promoting biosynthesis of amphotericin B, method for increasing amphotericin B production

Method used

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  • Method for producing amphotericin B through fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1、10

[0027] Embodiment 1, 10L automatic fermenter fermentation prepares amphotericin B

[0028] Preparation of Seed Solution of Streptomycin Nodosa Strain

[0029] Inoculate the slant culture or spore liquid of streptomycin nodosa strain on the seed medium, and cultivate at 26°C, 220 rpm, for 48 hours to obtain the seed liquid;

[0030] The seed culture medium described therein is prepared by the following method: 20.0 grams of cornstarch, 20.0 grams of glucose, 10.0 grams of hot-pressed soybean cake powder, 10.0 grams of yeast powder, 8.0 grams of corn steep liquor, 1.0 grams of sodium chloride, and 3.0 grams of calcium carbonate , add tap water to make up to 1000mL, pH 7.0, and sterilize at 121°C for 30min.

[0031] The amphotericin B producing bacterium of the present invention is a strain numbered ATCC14899.

[0032] Fermentation medium composition:

[0033] 280.0 grams of glucose, 140.0 grams of corn flour, 140.0 grams of hot-pressed soybean cake powder, 35.0 grams of corn ...

Embodiment 2、5

[0036] Embodiment 2, 5L automatic fermenter fermentation prepares amphotericin B

[0037] Preparation of Seed Solution of Streptomycin Nodosa Strain

[0038] Inoculate the slant culture or spore liquid of streptomycin nodosa strain into the seed medium, and cultivate for 46 hours at 28°C and 220 rpm to obtain the seed liquid;

[0039] The seed culture medium described therein is prepared by the following method: 30.0 grams of cornstarch, 10.0 grams of glucose, 15.0 grams of hot-pressed soybean cake powder, 8.0 grams of yeast powder, 5.0 grams of corn steep liquor, 2.0 grams of sodium chloride, and 4.0 grams of calcium carbonate , add tap water to make up to 1000mL, pH 7.5, and sterilize at 121°C for 30min.

[0040] The amphotericin B producing bacterium of the present invention is a strain numbered ATCC14899.

[0041] Fermentation medium composition:

[0042] Glucose 90.0g, corn flour 90.0g, cottonseed protein powder 90.0g, yeast powder 15.0g, corn steep liquor 18.0g, potas...

Embodiment 3、10

[0045] Embodiment 3, 10L automatic fermenter fermentation prepares amphotericin B

[0046] Preparation of Seed Solution of Streptomycin Nodosa Strain

[0047] Inoculate the slant culture or spore liquid of streptomycin nodosa strain on the seed medium, and cultivate for 44 hours at 30°C and 220 rpm to obtain the seed liquid;

[0048] The seed culture medium described therein was prepared by the following method: 25.0 grams of cornstarch, 15.0 grams of glucose, 20.0 grams of hot-pressed soybean cake powder, 5.0 grams of yeast powder, 2.0 grams of corn steep liquor, 1.0 grams of sodium chloride, and 3.0 grams of calcium carbonate , add tap water to make up to 1000mL, pH 7.0, and sterilize at 121°C for 30min.

[0049] The amphotericin B producing bacterium of the present invention is a strain numbered ATCC14899.

[0050] Fermentation medium composition:

[0051] Glucose 140g, corn flour 210g, cottonseed protein powder 175g, yeast powder 35g, corn steep liquor 56g, potassium di...

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Abstract

The invention belongs to the field of fermentation engineering, and in particular relates to a method for improving the yield of amphotericin B produced through microbial fermentation. According to the method, by optimizing organic complex nitrogen sources and the fermentation conditions, the influences of the nitrogen sources of different batches on the fermentation metabolism are reduced; by adding precusor substances, the biosynthesis of the amphotericin B is promoted. After the fermentation is finished, the fermentation unit of the amphotericin B achieves 14469ug / ml. Compared with the result before optimization, the yield of the amphotericin B is improved by 103%, the fermentation time is shortened by 14%, the utilization rate of equipment is improved, and the purification of the downstream is benefited.

Description

technical field [0001] The invention belongs to the field of fermentation engineering, in particular to a method for producing amphotericin B by fermentation. Background technique [0002] Amphotericin B is produced by the fermentation of streptomycin nodosa, a polyene macrolide antibiotic composed of 37 carbon atoms, and is clinically used to treat a variety of deep fungi such as Candida albicans, Cryptococcus neoformans, Heart or systemic infection caused by Aspergillus, etc. Treatment of certain severe deep mycoses such as Cryptococcal neoformans meningitis, invasive aspergillosis, especially in immunocompromised or severe agranulocytosis patients and certain endemic mycoses such as coccidioidomycosis, histoplasmic Amphotericin B is still required for mycosis, dermatitis blastomycosis, etc. Amphotericin B is still the first-line drug and the "gold standard" for the treatment of invasive fungal infections. [0003] Amphotericin B is a polyene macrolide antibiotic, and it...

Claims

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Application Information

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IPC IPC(8): C12P19/62
CPCC12P19/62
Inventor 段宝玲张雪霞王耀耀任风芝李宁耿文飞高任龙郑智慧路新华
Owner NCPC NEW DRUG RES & DEV
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