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Method for producing sisomicin by streptomyces fradiae variant GB-2

A technology of Streptomyces freundii and sisomycin, applied in the direction of microorganism-based methods, biochemical equipment and methods, chemical instruments and methods, etc., to save time and money

Inactive Publication Date: 2009-01-07
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

At present, there is no report at home and abroad that other strains other than the above can produce sisomycin

Method used

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  • Method for producing sisomicin by streptomyces fradiae variant GB-2
  • Method for producing sisomicin by streptomyces fradiae variant GB-2
  • Method for producing sisomicin by streptomyces fradiae variant GB-2

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Embodiment Construction

[0031] Streptomyces flexneri variant GB-2 produces sisomycin fermentation and isolation identification method as follows:

[0032] 1) Fermentation of sisomycin produced by Streptomyces flexneri variant GB-2

[0033] Streptomyces freundii GB-2 (Liu Shu et al., Journal of Nanjing Agricultural University, 2007, 4: 124-129) was inoculated into Gao's synthetic No. 1 medium (Zheng Z H, et al. FEMS Microbiol Lett, 2000, 188: 67-91) on the slant, cultured at 28°C for 7 days. Take the freshly cultivated GB-2 slant and wash the spores with 50mL sterile water containing glass beads, 180r min -1 Shake for 20 minutes to make a bacterial suspension. Inoculate the bacterium suspension into the fermentation medium (soybean powder medium: 2.0 g of soluble starch, 1.5 g of soybean powder, 0.5 g of glucose, 0.25 g of yeast extract, CaCO 3 0.1g, in 1000mL of artificial seawater, pH7.5), at 28℃180r·min -1 Culture 7d. Collect fermentation broth, 5000r·min -1 Centrifuge for 15 minutes to remo...

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Abstract

The invention relates to a method for producing sisomicin sulfate by the variation GB-2 of Freund streptomyces and belongs to the technical field of microorganism fermentation. The variation GB-2 is inoculated into a fermentation culture liquid and cultured for 7 day under the temperature of 28 DEG C with a speed of 180r.min<-1>; a permeate liquid equal to or less than 1KDa obtained by carrying out centrifugating and hyperfiltration on a supernatant fluid is pre-treated and then goes on a column of Sephadex LH-20, collects the liquid and detects the antibacterialactivity, then a second apex which is namely the product of sisomicin sulfate is collected after effective liquidoid chromatogram analyzing and HPLC purifying are carried out. Detected by ESI-MS, the molecular weight thereof is m / z 447.7 and is confirmed to be the sisomicin sulfate by a secondary mass spectrum. The method of the invention firstly obtains the sisomicin sulfate in the Freund streptomyces which belongs to the streptomyces of actinomycetes. The sisomicin sulfate is named to be the aminoglycoside antibiotic of the third generation which is the antibiotic with the advantages of the smallest side effect, strongest resistance of pseudomonas aeruginosa, broadest capacity for resisting modifying enzyme and broadest antimicrobial spectrum in the aminoglycoside antibiotics and has a potential market application prospect.

Description

1. Technical field [0001] The invention relates to a method for producing sisomycin by Streptomyces flexneri variant GB-2, which belongs to the technical field of microbial fermentation, and is specially used for microbial fermentation, separation and identification of sisomycin produced by Streptomyces flexneri variant GB-2. 2. Background technology [0002] Micromonospora microorganisms in actinomycetes can produce many important aminoglycoside antibiotics, such as gentamicin, sisomicin and so on. Sithomycin can be produced by Micromonospora rosea and Micromonospora inyoensis. Sithomycin is an aminoglycoside broad-spectrum antibiotic, which has the least side effects, the strongest resistance to Pseudomonas aeruginosa, and the broadest resistance to inactivating enzymes among aminoglycoside antibiotics. At present, there is no report at home and abroad that other strains other than the above can produce sisomycin. Our research group screened out a strain of marine Strept...

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Application Information

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IPC IPC(8): C12P19/48C07H15/236C07H1/06C12R1/54
Inventor 陆颖健别小妹董昕刘姝吕凤霞
Owner NANJING AGRICULTURAL UNIVERSITY
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