High efficiency fermentation method for pristinamycin

A technology of protomycin and fermentation culture, applied in the direction of fermentation, bacteria, etc., can solve the problems of low fermentation titer and high production cost, and achieve the effect of increasing fermentation titer

Inactive Publication Date: 2005-04-20
ZHEJIANG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to overcome the disadvantages of low fermentation titer and high production cost of existing protomycin, the invention provides a Streptomyces pristinaespiralis microorganism

Method used

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Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0073] Preparation of spore culture:

[0074] Spore culture medium: adopt Gao's No. 1 medium (20 grams of soluble starch, KNO 3 1 gram, K 2 HPO 4 0.5 g, NaCl 0.5 g, MgSO 4 0.5 g, FeSO 4 0.01 g, 20 g agar, add distilled water to 1 liter, adjust the pH to 7.0-7.2 with sodium hydroxide before the medium is sterilized). The above medium components were mixed and distributed into small test tubes. After sterilizing at 121° C. for 30 minutes, spread the medium in the test tubes on a slope.

[0075] Streptomyces erotiscensus was inoculated on the spore-forming medium and cultured for 6 days at a temperature of 30°C.

[0076] Seed preparation:

[0077] Seed medium: 30 grams of glucose, 10 grams of soybean cake powder, 5 grams of peptone, 5 grams of yeast powder, KNO 3 3 g, NaCl 2 g%, CaCO 3 4 g, add tap water to 1 liter, adjust pH to 7.0 with sodium hydroxide before sterilization.

[0078] Preparation of seed culture medium: After the seed culture medium was mixed, it was div...

experiment example 2

[0086] Preparation of seeds in 110 liter seed pots:

[0087] Pack 60 liters of seed culture medium in 110 liters of seed tanks, and the composition of the culture medium is the same as that of the shaking flask seed culture medium in Experimental Example 1, but 26 milliliters of distilled water will be added. The medium was sterilized at 121° C. for 30 minutes by the solid tank sterilization method.

[0088] 100 milliliters of shake flask seeds were inoculated into 110 liters of seed pots containing 60 liters of seed medium by flame method to cultivate and prepare seeds. Oxygen is provided in the seed tank by feeding sterile compressed air and stirring (the ventilation rate is 0.8 liters of air per liter of seed solution per minute, and the stirring speed is 200 rpm). Keep the seed culture temperature at 30°C and cultivate for 36 hours. The cultivation was terminated when the concentration of the mycelia of Streptomyces involutes in the seed solution reached 20%.

[0089] P...

experiment example 3

[0094] 700 liters of fermentation broth (3500u / ml) obtained in Experimental Example 2 was adjusted to pH 3 with 6N hydrochloric acid, filtered, and top washed with 100 liters of water. The filtrate was adjusted to pH 7 with dilute sodium hydroxide solution, and then extracted with dichloromethane. The organic phase was concentrated to 4.8 L under reduced pressure. 5 liters of petroleum ether was added to the concentrated solution to obtain a precipitate, which was washed and dried to obtain 236 grams of a crude product with a potency of 7560 u / mg.

[0095] Dissolve 236 grams of crude product (7560u / mg) in 12 liters of dichloromethane, filter, and the filtrate enters a column containing 3.6 liters of granular activated carbon, rinses with 22 liters of dichloromethane, collects the central part, concentrates under reduced pressure to 1.2 liters, and concentrates Liquid was filtered, and the filtrate was treated with petroleum ether to obtain a precipitate. After washing and dryin...

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PUM

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Abstract

Said invention relates to Streptomyces pristinaespiralis CGMCC 0957 and production and method of pristinamycin by same, which is that a method for high effectively producing Streptomyces pristinaespiralis CGMCC 0957 in culture medium under proper condition.

Description

Technical field: [0001] The invention relates to a method for fermenting Streptomyces initiosa CGMCC 0957 to efficiently produce protomycin. Background technique: [0002] Pristinamycin belongs to the streptavidin antibiotics and consists of about 30% pristinamycin I and about 70% pristinamycin II. Protomycin I is a cyclic hexapeptide lactone derived from A , Protomycin I B and primitive mycin I C composition. Protomycin I A The molecular weight is 866, and the molecular formula is C 45 h 54 o 10 N 8 ; Protomycin I B The molecular weight is 852, and the molecular formula is C 44 h 52 o 10 N 8 ; Protomycin I C The molecular weight is 852, and the molecular formula is C 44 h 52 o 10 N 8 . Protomycin II is a polyunsaturated cyclic lactone derived from Protomycin II A and primitive mycin II B composition. Protomycin II A The molecular weight is 525, and the molecular formula is C 28 h 35 o 7 N 3 ; Protomycin II B The molecular weight is 527, and the m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P17/18C12P21/02
Inventor 金志华李春波岑沛霖张国钧金一平陶正利林建平徐志南
Owner ZHEJIANG UNIV
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