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Levo-phosphonomycin biotransformation strain screening method using dextro-phosphonomycin as substrate

A technology of D-fosfomycin and L-fosfomycin, applied in the direction of biochemical equipment and methods, microbial determination/inspection, etc.

Active Publication Date: 2012-11-14
SHENYANG INST OF APPL ECOLOGY CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This not only leads to increased production costs and waste of resources, but also creates serious environmental pollution problems

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0036] 1. Preparation of complete liquid medium and strain cultivation

[0037] In terms of weight percentage, the complete liquid medium has the following formula: 0.5% beef extract, 1% peptone, 0.5% NaCl, the balance is distilled water, pH 7.2-7.4, sterilized at 115°C for 30 minutes, and set aside.

[0038]Inoculate the strain to be tested in a complete liquid medium for enrichment at 30°C, shake the shaker at 160rpm, and shake for 3 days. Compared with the blank control complete liquid medium, it is obviously turbid with the naked eye, and then add the conversion substrate D-phosphorus Mycin, so that the weight ratio is 0.1 to 0.8%, and adding trace element NaVO 3 and CoCl 2 , so that the weight ratios were 0.02% and 0.05%, respectively, and carried out transformation culture at 30° C. for 5 days, shaking on a shaker at 160 rpm, to obtain a fermented liquid after substrate transformation.

[0039] 2. Product identification

[0040] 1. Indicator bacteria culture and suspe...

Embodiment

[0051] The strains screened from the soil by the strain screening model using cPA as the only carbon source in the laboratory in the early stage were enriched and cultured in the early stage, and the Rf value was compared with the subsequent biological assay and TLC test, and the strains with the following right Levofosfomycin biotransformation strain as substrate.

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PUM

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Abstract

The invention relates to biological transformation / biocatalytic technology microbial bacterial strain sieving domain, especially to a sieving policy and method for left-handed fosfomycin biological converting bacterial strain with right-handed fosfomycin as substrate. The said sieving policy and specific method of operation consist of two big steps including bacterial strain culture and conversion product identification in screening process, in which the essential point includes preliminary enrichement of laboratory bacterial and product calibration policy which combines biological calibration with thin-layered chromatography (TLC). Because the bacterial which can be changed from right-handed fosfomycin to left-handed fosfomycin in nature is rare and the lab available bacterial grows up slowly, the preliminary enrichement of laboratory bacterial is very important. Moreover, adopting product calibration policy which combines biological calibration with thin-layered chromatography (TLC), whether there is left-handed fosfomycin in product can be exactly determined.

Description

technical field [0001] The invention relates to the field of biotransformation / biocatalysis technology and screening of microbial strains, in particular to a strategy and method for screening and testing of levofosfomycin biotransformation strains using levofosfomycin 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, fosfomyci...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04
Inventor 张雪姝徐慧张颖陈冠雄陈玉张海燕
Owner SHENYANG INST OF APPL ECOLOGY CHINESE ACAD OF SCI
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