New screen method of levofosfomycin strain converted by d-phosphonomycin as substrate

Abstract

The invention relates to the technical field of biotransformation / biocatalysis, and relates to a novel screening method for levo-fosfomycin biotransformation strains with dextro-fosfomycin as the substrate. Solid medium containing the dextro-fosfomycin is utilized to culture strains to be tested; oversensitive micrococcus luteus and super-tolerance colon bacillus are utilized to be as the bioassay indicator bacteria, an agar block is placed on a bioassay tray to screen out levo-fosfomycin ransformation strains, then the products are identified by the combined method of the thin layer chromatography and the biological developer method to obtain positive strains to undergo complete liquid medium transformation with the dextro-fosfomycin as the substrate, transformation liquid is concentrated to undergo the product identification by the combined method of the thin layer chromatography and the biological developer method, and finally levo-fosfomycin biotransformation dominant strains are screened out. The method has low requirements for equipment, simple operation and low cost, and is suitable for the batch screening of the biotransformation bacteria.

Description

Technical field: [0001] The invention relates to the technical field of biotransformation / biocatalysis, in particular to a screening method for a new levofosfomycin biotransformation bacterial strain using levofosfomycin as a substrate. Background technique: [0002] Fosfomycin [(-)-cis-(1R, 2S)-1,2-epoxypropyl phosphate, English name: Fosfomycin, referred to as FOM] is a clinically important broad-spectrum antibiotic that inhibits the synthesis of bacterial cell walls. It was discovered in Streptomyces freundii by Hendlin et al. in 1969. It covalently binds to pyruvyltransferase, causing irreversible inactivation of the enzyme and inhibiting the synthesis of N-acetylmuramic acid. Fosfomycin has a simple chemical structure, unique antibacterial mechanism, wider antibacterial spectrum than β-lactam antibiotics, similar to aminoglycoside antibiotics gentamicin and tobramycin, and no cross-resistance to other antibiotics , and most of them show a synergistic effect. It has a...

AI Technical Summary

Problems solved by technology

Because the yield of the first step chemical epoxidation is 70%, the second step chemical resolution yield is less than 50%, the total yield of producing levofosfomycin by chemical synthesis method is less than 35% like this, and the total yield of chemical synthesis process The rate is less than 20%, so people have been hoping to find other more effective ways

[0004] Since the 1970s, people have begun to study the asymmetric synthesis of levofosfomycin, among which there are many studies on the direct production of levofosfomycin by microbial transformation, but they are all limited to cis-propenylphosphonic acid , cPA) as the substrate for biotransformation, so far, there is no research report on the successful conversion of fosfomycin into levofosfomycin at home and abroad with the waste of chemically synthesized fosfomycin-dexfosfomycin as the substrate