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Preliminary Studies for Cephamycin C Purification Technique

Author:
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
Source:
Applied biochemistry and biotechnology 2012 v.166 no.1 pp. 208-221
ISSN:
0273-2289
Subject:
Escherichia coli, Streptomyces clavuligerus, adsorption, antibiotics, fermentation, ion exchange chromatography, mass spectrometry, microfiltration, nuclear magnetic resonance spectroscopy, purification methods, resins, ultrafiltration
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 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.
Agid:
433020