Method for screening biological catalyst strain for converting levo-fosfomycin into dextral-fosfomycin

A technology of levofosfomycin and dexfofomycin, which is applied to biochemical equipment and methods, microorganisms, microorganisms, etc., and can solve the problems of screening failures and large numbers of omissions

Inactive Publication Date: 2011-08-17
SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI +1
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Problems solved by technology

[0006] The purpose of the present invention is to provide a new screening method for transforming D-fosfomycin into Levofosfomycin catalytic strains, to solve the problems in the prior art tha

Method used

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  • Method for screening biological catalyst strain for converting levo-fosfomycin into dextral-fosfomycin

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Embodiment

[0027] First, inoculate the beef extract peptone liquid medium with Escherichia coli, cultivate it at 37°C for 24 hours, and prepare the bacterial suspension of Escherichia coli strain, so that the bacterial concentration is 10 8 -10 10 bacteria / ml, which is made into a high-concentration sensitive bacteria suspension.

[0028] Then the catalytic bacterial strain to be screened may be transformed into levofosfomycin (through these two characteristics of levofosfomycin resistance and / or dextrofosfomycin utilization, use conventional techniques to preliminarily judge whether the bacterial strain is Potentially transformable bacterial strains) were inoculated in the catalytic medium, and after 72 hours of shaking flask culture, the fermentation broth was drawn, and the bacteria in the fermentation broth were removed by filtration through a 0.45 μm pore size filter membrane to prepare a sterile fermentation broth of the bacterial strain to be screened.

[0029] Then, mix the abov...

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Abstract

The invention relates to the technical field of biomedicines and biology, in particular to a high-sensitivity method for screening a strain for catalyzing the conversion of levo-fosfomycin into dextral-fosfomycin, which comprises: firstly, preparing bacterial suspension of a dextral-fosfomycin sensitive strain, wherein the bacterial concentration is 10<8> to 10<10>/ml; secondly, adding the bacterial suspension of the dextral-fosfomycin sensitive strain into sterile fermentation liquid of catalyst strain to be screened, which is filtered by a filter membrane with an aperture of 0.45 mu m to remove bacteria, collecting fermentation liquid at the starting time of culture and after a certain period of culture, and measuring the absorbance of the collected fermentation liquid by using a 600-nanometer spectrophotometer; and selecting fermentation liquor of which the absorbance increases the least or less after a period of culture, and further verifying if the original strain to be screened has the capability of converting levo-fosfomycin into dextral-fosfomycin. When the method is used, the sensitivity of the method for screening the biological catalyst strain for converting levo-fosfomycin into dextral-fosfomycin is improved greatly, and the screening efficiency is improved greatly for the detection method is simple and easy to implement.

Description

technical field [0001] The invention relates to the fields of biomedicine and biotechnology, in particular to a highly sensitive screening method for catalyzing D-fosfomycin to L-fosfomycin bacterial strains. Background technique [0002] Levofosfomycin [(-)-cis-(1R,2S)-1,2-epoxypropyl phosphate, here abbreviated as FOM] is a broad-spectrum antibiotic. It has a unique chemical structure and antibacterial mechanism, not only has no cross-resistance with other antibiotics, but also mostly has a synergistic effect. At the same time, because fosfomycin also has the advantages of stable physical and chemical properties, safety, low toxicity, and no antigenicity, it has been widely used in clinical treatment and is called a new antibiotic in the 21st century by the international medical and pharmaceutical circles. [0003] At present, what levofosfomycin production technique adopts is chemical synthesis method, and this technique yield is lower (less than 20%), and the main reaso...

Claims

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Application Information

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IPC IPC(8): C12N1/00G01N21/31
Inventor 徐慧杨伟超王若超周丽沙韩斯琴孔双张大鹏纪悦周凯赵华季光辉金玉坤
Owner SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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