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Method for screening levo-phosphonomycin bioconversion strain with cis-propene phosphoric acid as substrate

A technology of cis-acrylphosphate and levofosfomycin, which is applied in the direction of biochemical equipment and methods, microorganisms, microorganisms, etc., can solve the problems of low tolerance and transformation ability that cannot reach industrial production, so as to improve the overall efficiency and reduce complex The effect of screening workload and easy operation

Inactive Publication Date: 2005-09-28
SHENYANG INST OF APPL ECOLOGY CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
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AI Technical Summary

Problems solved by technology

However, because these strains have very low tolerance to substrates and products, their transformation capabilities are far from meeting the needs of industrial production.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0045] Preparation of agar blocks: Melt the above medium, pour 25mL into each 90mm diameter plate, and let it stand to solidify. After solidification, punch out the agar block (diameter 6mm, height 8mm), then inoculate the bacterial strain to be tested obtained in the preliminary screening step on its surface, and place it at 30° C. and cultivate it under 75%-85% relative humidity for 4-5 days. sky.

[0046] 2. Identification of fosfomycin transformed strains

[0047] Preparation of bioassay media and their plates:

[0048] Medium I composition: by weight percentage, beef extract 0.5%, peptone 1%, NaCl 0.5%, agar 2%, balance is distilled water, pH 7.0-7.2; Medium II composition: by weight percentage, beef extract 0.5 %, peptone 1%, NaCl 0.5%, agar 1.5%, the rest is distilled water, pH7.0-7.2; medium I and medium II were sterilized at 121°C for 20 minutes. Pour 200mL of melted medium I into a biological identification tray (20×30cm) to make a lower plate; then cool 100mL of ...

Embodiment

[0056] Five kinds of soil samples collected from different sites were made into soil suspension, and more than 180 strains of bacteria were obtained after the initial screening step. After these strains were re-screened, more than 30 strains of bacteria were found to have obvious antibacterial effect lock up. Through thin-layer chromatography test, it is proved that these strains present a bacteriostatic zone by transforming cis-acrylic acid into levofosfomycin.

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PUM

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Abstract

The present invention relates to biological conversion / biological catalysis technology and microbe strain screening technology, and is especially the screening police and method of levorotary phosphonomycin bioconverting strain with maleic phosphoric acid as substrate. The screening process includes two steps of initial screening and re-screening. In the initial screening, the selective culture medium has maleic phosphoric acid as only carbon source so as to eliminating most of irrelative microbes. In the re-screening, the strain obtained in the initial screening and the solid culture medium with levorotary phosphonomycin bioconverting strain and maleic phosphoric acid substrate are adopted to sort out the phosphonomycin bioconverting strain. The present invention has greatly raised screening efficiency.

Description

technical field [0001] The invention relates to the fields of biotransformation / biocatalysis technology and screening of microbial strains, in particular to a screening strategy and method for levofosfomycin biotransformation strains using cis-allylphosphate as a substrate. Background technique [0002] Fosfomycin [(-)-cis-(1R,2S)-1,2-epoxypropyl phosphate, English name: Fosfomycin, FOM for short] is a broad-spectrum antibiotic. Fosfomycin has a simple chemical structure and is quite different from most commonly used antibiotics. It has the characteristics of low toxicity, no antigenicity, wide antibacterial spectrum, multiple indications, and is not easy to form drug resistance. The results of research in recent years have discovered some new effects of fosfomycin, such as: synergistic effect with other antibiotics, reduction of toxic and side effects of other antibiotics, combined drug treatment of severe infections, and immune enhancement. Therefore, fosfomycin is called...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/00C12Q1/04
Inventor 徐慧李智峰张雪姝张颖陈冠雄张忠泽纪悦王兰君陈玉
Owner SHENYANG INST OF APPL ECOLOGY CHINESE ACAD OF SCI
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