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Preliminary Studies for Cephamycin C Purification Technique

de Baptista Neto, Álvaro, Bustamante, Maritza Catalina Condori, de Oliveira, Jaine Honorata Hortolan Luiz, Granato, Ana Cláudia, Bellão, Carolina, Junior, Alberto Colli Badino, Barboza, Marlei, Hokka, Carlos Osamu
Applied biochemistry and biotechnology 2012 v.166 no.1 pp. 208-221
Escherichia coli, Streptomyces clavuligerus, adsorption, antibiotics, fermentation, ion exchange chromatography, mass spectrometry, microfiltration, nuclear magnetic resonance spectroscopy, purification methods, resins, ultrafiltration
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 β-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.