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Method for producing capreomycin by fermentation

The technology of capreomycin and Streptomyces crispatus is applied in the field of fermentation engineering, which can solve problems such as low fermentation level, achieve the effects of increasing fermentation unit, reducing feedback inhibition, and avoiding strain poisoning

Inactive Publication Date: 2016-04-20
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 low fermentation level in the preparation of capreomycin by microbial fermentation in the prior art, and to provide a method of optimizing the fermentation medium formula and fermentation conditions, adding precursor substances in the fermentation process, and promoting the production of capreomycin. biosynthesis, thereby improving the method of capreomycin production

Method used

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  • Method for producing capreomycin by fermentation

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Experimental program
Comparison scheme
Effect test

Embodiment 1、5

[0027] Embodiment 1, 5L automatic fermenter fermentation prepares capreomycin

[0028] Seed solution of Streptomyces crimperidus strain

[0029] Inoculate the slant culture or spore liquid of the Streptomyces crispatus strain into the seed medium, and cultivate for 72 hours at 26°C and 220 rpm to obtain the seed liquid;

[0030] The seed culture medium described therein is prepared as follows: 20.0 grams of sucrose, 10.0 grams of yeast powder, 2.0 grams of ammonium sulfate, 2.0 grams of corn steep liquor, 0.5 grams of sodium chloride, 4.0 grams of calcium carbonate, add tap water to 1000mL, pH6.5, sterilized at 121°C for 30min.

[0031] The capreomycin-producing bacterium of the present invention is a strain numbered NRRL2773.

[0032] Fermentation medium composition:

[0033] 60.0 grams of corn starch, 60.0 grams of soybean cake powder, 12.0 grams of ammonium sulfate, 12.0 grams of corn steep liquor, 12.0 grams of potassium dihydrogen phosphate, 6.0 grams of sodium chlorid...

Embodiment 2、2

[0036] Embodiment 2, 2L automatic fermenter fermentation preparation capreomycin

[0037] Seed solution of Streptomyces crimperidus strain

[0038] Inoculate the slant culture or spore liquid of the Streptomyces crispatus strain into the seed medium, and cultivate for 64 hours at 28°C and 220rpm to obtain the seed liquid;

[0039] The seed culture medium described therein is prepared as follows: 40.0 grams of sucrose, 5.0 grams of yeast powder, 3.0 grams of ammonium sulfate, 8.0 grams of corn steep liquor, 1.0 grams of sodium chloride, 3.0 grams of calcium carbonate, add tap water to 1000mL, pH7.0, sterilized at 121°C for 30min.

[0040] The capreomycin-producing bacterium of the present invention is a strain numbered NRRL2773.

[0041] Fermentation medium composition:

[0042]Corn starch 30.0g, cottonseed protein powder 20.0g, yeast powder 5.0g, ammonium sulfate 2.0g, corn steep liquor 8.0g, potassium dihydrogen phosphate 2.0g, sodium chloride 1.0g, calcium carbonate 6.0g,...

Embodiment 3、2

[0045] Embodiment 3, 2L automatic tank fermentation prepares capreomycin

[0046] Seed solution of Streptomyces cristiformis strain

[0047] Inoculate the slant culture or spore liquid of the Streptomyces crispatus strain into the seed medium, and cultivate for 48 hours at 30°C and 220 rpm to obtain the seed liquid;

[0048] The seed culture medium described therein was prepared as follows: 30.0 grams of sucrose, 8.0 grams of yeast powder, 2.0 grams of ammonium sulfate, 4.0 grams of corn steep liquor, 2.0 grams of sodium chloride, 6.0 grams of calcium carbonate, added tap water to 1000mL, pH6.5, sterilized at 121°C for 30min.

[0049] The capreomycin-producing bacterium of the present invention is a strain numbered NRRL2773.

[0050] Fermentation medium composition:

[0051] Corn starch 30.0g, cottonseed protein powder 20.0g, yeast powder 5.0g, ammonium sulfate 2.0g, corn steep liquor 8.0g, potassium dihydrogen phosphate 2.0g, sodium chloride 1.0g, calcium carbonate 6.0g, a...

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Abstract

The invention belongs to the field of fermentation engineering and particularly relates to a method for producing capreomycin by fermentation. The method ensures stability of inter-batch fermentation units by optimizing a culture medium and fermentation conditions; by adding precursor material in a fermentation process, production of capreomycin can be significantly promoted; by adding super-paramagnetic microspheres in a capreomycin synthesis phase to adsorb capreomycin in a fermentation broth, feedback inhibition of a fermentation product upon generated bacteria is lowered. After fermentation, fermentation unit of the capreomycin is up to 14149 ug / ml; compared with pre-optimization methods, this method provides an increase of 73% in capreomycin yield, an improvement in productivity and a reduction in energy consumption and waste discharge.

Description

technical field [0001] The invention belongs to the field of fermentation engineering, in particular to a method for producing capreomycin by fermentation. Background technique [0002] The resurgence of tuberculosis is seriously threatening human health. In recent years, the emergence of multidrug-resistant tuberculosis (MDR-TB) and the co-infection of Mycobacterium tuberculosis and human immunodeficiency virus have led to the re-emergence of tuberculosis in the world. Capreomycin is an important class of drugs for the treatment of multidrug-resistant Mycobacterium tuberculosis and persistent bacterial infections. The drug has a remarkable curative effect on pulmonary tuberculosis caused by drug-resistant Mycobacterium tuberculosis infection, and has less toxic and side effects than other second-line drugs, and is currently an ideal anti-TB drug. [0003] Capreomycin is a cyclic polypeptide antibiotic produced by the fermentation of Streptomyces crispatus. It is composed ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/04C12R1/465
CPCC12P21/02C12N1/205C12R2001/465
Inventor 高任龙王耀耀张雪霞任风芝耿文飞路新华段宝玲杨海静
Owner NCPC NEW DRUG RES & DEV
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