Preliminary Studies for Cephamycin C Purification Technique
2012
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
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.
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