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Appl Biochem Biotechnol ; 166(1): 208-21, 2012 Jan.
Article in English | MEDLINE | ID: mdl-22057909

ABSTRACT

A study was made for purification of cephamycin C from fermentation of Streptomyces clavuligerus. Initially, the culture broth was clarified by microfiltration and ultrafiltration, after which the resulting permeates were subjected to nonspecific adsorption and ion-exchange chromatography on resin columns. The antibiotic activity was measured by the biological method at each stage by assaying its activity against the Escherichia coli ESS, super sensitive to ß-lactam antibiotic. The purification processes were assessed in relation to the variables affecting each step. The purification efficiency by nonspecific adsorption was monitored by UV spectrophotometry, while the ion-exchange adsorption fractions were assessed by NMR spectroscopy. Some of the fractions obtained during purification were also analyzed by mass spectrometry (LC/MS and LC/MS/MS) to identify the cephamycin C molecule. These preliminary results proved the process feasibility.


Subject(s)
Anti-Infective Agents/chemistry , Anti-Infective Agents/isolation & purification , Cephamycins/chemistry , Cephamycins/isolation & purification , Streptomyces/chemistry , Anti-Infective Agents/pharmacology , Cephamycins/pharmacology , Chromatography, Ion Exchange , Escherichia coli/drug effects , Fermentation , Magnetic Resonance Spectroscopy , Mass Spectrometry , Recombinant Proteins , Spectrophotometry, Ultraviolet , beta-Lactams/chemistry
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