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Method for producing Rakicidins compounds by virtue of marine micromonospora fermentation

A compound and fermenter technology, applied in the field of industrial microorganisms and biopharmaceuticals, can solve the problems of low fermentation titer and inability to achieve industrialized production, and achieve the effect of meeting the needs of industrialization

Active Publication Date: 2016-07-20
FUJIAN INST OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to overcome the defects that the fermentation titer of Rakicidins compounds in the prior art is low and cannot meet the technical requirements of industrialized production

Method used

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  • Method for producing Rakicidins compounds by virtue of marine micromonospora fermentation
  • Method for producing Rakicidins compounds by virtue of marine micromonospora fermentation
  • Method for producing Rakicidins compounds by virtue of marine micromonospora fermentation

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Micromonospora marine (Micromonospora sp. FIM02-523) spores preserved on a solid slope were scraped and inoculated into a shake flask primary seed culture by scraping the spores, and cultured at 28° C. and 220 rpm for 48 hours.

[0037] Seed medium formula (%): soluble starch 2.0, glucose 1.0, yeast powder 2.0, peptone 1.0, MgSO 4 ·7H 2 O0.05, FeSO 4 ·7H 2 O0.005, CuSO 4 ·5H 2 O0.005, CoCl 2 ·6H 2 O0.0005, CaCO 3 0.1~0.3, prepared with tap water, pH, 7.0~7.5; 28~32℃.

[0038] Then inoculate 5% inoculum in a 20L fermenter (actual liquid volume 15L) for fermentation, culture at 28°C, start at 200rpm, adjust to 350rpm after 48 hours, and finish fermentation for about 96-120 hours. The tank pressure is 0.03-0.05Mpa, and the ventilation volume is 1:1vvm.

[0039] The fermentation medium is: soluble starch 5.0, sucrose 1.0, soybean cake powder 3.0, yeast powder 1.0, MgSO 4 ·7H 2 O0.04, FeSO 4 ·7H 2 O0.005, CuSO 4 ·5H 2 O0.005, CoCl 2 ·6H 2 O0.0005, CaCO 3 0....

Embodiment 2

[0041] The basic conditions of the fermentation process are the same as in Example 1, except that 0.2% methionine in the volume content of the fermentation broth is added at one time after 24 hours of fermentation. During the fermentation process, the content of Rakicidins compounds in the fermentation broth was detected by HPLC. The final fermentation titer was 270mg / L, wherein the content of RakicidinA was 50mg / L, the content of RakicidinB1 was 120mg / L, and the content of RakicidinB was 110mg / L.

Embodiment 3

[0043] The basic conditions of the fermentation process are the same as in Example 1, except that 0.3% glycine in the volume content of the fermentation broth is added at one time after 24 hours of fermentation. During the fermentation process, the content of Rakicidins compounds in the fermentation broth was detected by HPLC, and the final fermentation titer was 570mg / L, wherein the content of RakicidinA was 140mg / L, the content of RakicidinB1 was 80mg / L, and the content of RakicidinB was 350mg / L.

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Abstract

The invention relates to the technical field of industrial microorganisms and the field of biological medicines, and in particular relates to a method for producing Rakicidins compounds by virtue of marine micromonospora fermentation. The method is characterized by comprising the following steps: performing solid slant culture on micromonospora which is collected in the China General Microbiological Culture Collection Center (CGMCC) and has a collection number of CGMCC NO. 12132, performing shake flask seed culture, seeding tank culture and fermentation tank fermentation culture, and performing after-treatment to obtain the compounds, wherein amino acid is replenished in the fermentation initial stage of a fermentation tank. The fermentation method provided by the invention is simple in operation and low in cost and can be used for realizing industrial large-scale production, and the highest total fermentation titer reaches about 600mg / L.

Description

technical field [0001] The invention relates to the technical field of industrial microbes and the field of biomedicine, in particular to a method for fermenting Micromonospora marine to produce Rakicidins compounds. Background technique [0002] The anti-cancer drug market has grown rapidly in recent years. In 2014, the global sales of anti-cancer drug market reached 100 billion US dollars; by 2018, the anti-cancer drug market will reach 147 billion US dollars, with a compound growth rate of 11.6%. The research and development of anti-cancer drugs will make huge profits . Hypoxia is one of the characteristics of malignant solid tumors. Hypoxia is closely related to tumor angiogenesis, invasion and metastasis, resistance to radiotherapy and chemotherapy, and poor prognosis. Hypoxia-inducible factor 1 (hypoxia-inducible factor 1, HIF-1) is the most critical nuclear transcriptional regulator in the regulation of hypoxia effects. HIF-1 is selectively and continuously highly e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P21/04C12R1/29
CPCC12P21/02C12N1/205C12R2001/29
Inventor 周剑林风江红连云阳江宏磊方东升陈丽赵薇
Owner FUJIAN INST OF MICROBIOLOGY
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