Preliminary Studies for Cephamycin C Purification Technique

Neto, Álvaro de Baptista; Bustamante, Maritza Catalina Condori; Luiz, Jaine Honorata Hortolan; Granato, Ana Cláudia; Bellão, Carolina; Badino, Alberto Colli; Barboza, Marlei; Hokka, Carlos O.

doi:10.1007/s12010-011-9417-6

Cited by 6 publications

(9 citation statements)

References 13 publications

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“…A calibration curve was performed using ten cephalosporin C concentration values from 5 to 120 mg l -1 and 24 replicates for each concentration. Antibiotic analyses were also carried out via high-performance liquid chromatography as described in Baptista Neto et al [37]. Lysine and alpha-aminoadipic acid analyses were conducted by means of the post-column derivatization method with orthophtalaldehyde and quantified in a fluorescence detector [38].…”

Section: Methodsmentioning

confidence: 99%

Enhancing effect of lysine combined with other compounds on cephamycin C production in Streptomyces clavuligerus

2013

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BackgroundLysine plays an important role in Streptomyces clavuligerus metabolism; it takes part in its catabolism, via cadaverine, and in its secondary metabolism, in which lysine is converted via 1-piperideine-6-carboxylate to alpha-aminoadipic acid, a beta-lactam antibiotic precursor. The role of lysine as an enhancer of cephamycin C production, when added to production medium at concentrations above 50 mmol l-1, has already been reported in the literature, with some studies attributing a positive influence to multifunctional diamines, among other compounds. However, there is a lack of research on the combined effect of these compounds on antibiotic production.ResultsResults from experimental design-based tests were used to conduct response surface-based optimization studies in order to investigate the synergistic effect of combining lysine with cadaverine, putrescine, 1,3-diaminopropane, or alpha-aminoadipic acid on cephamycin C volumetric production. Lysine combined with cadaverine influenced production positively, but only at low lysine concentrations. On the whole, higher putrescine concentrations (0.4 g l-1) affected negatively cephamycin C volumetric production. In comparison to culture media containing only lysine as additive, combinations of this amino acid with alpha-aminoadipic acid or 1,3-diaminopropane increased cephamycin C production by more than 100%.ConclusionThis study demonstrated that different combinations of lysine with diamines or lysine with alpha-aminoadipic acid engender significant differences with respect to antibiotic volumetric production, with emphasis on the benefits observed for lysine combined with alpha-aminoadipic acid or 1,3-diaminopropane. This increase is explained by mathematical models and demonstrated by means of bioreactor cultivations. Moreover, it is consistent with the positive influence of these compounds on lysine conversion to alpha-aminoadipic acid, a limiting step in cephamycin C production.

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Section: Methodsmentioning

confidence: 99%

Enhancing effect of lysine combined with other compounds on cephamycin C production in Streptomyces clavuligerus

2013

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“…Studies on the purification of cephamycin C demonstrated the presence of other compound (or compounds), besides cephamycin C, in fermentation broth and that are detected by the bioassay method [13]. Rollins et al [16] reported the production of β-lactam compounds by the biosynthetic pathway of cephamycin C production in conditions of dissolved oxygen limitation.…”

Section: Resultsmentioning

confidence: 99%

“…Rollins et al [16] reported the production of β-lactam compounds by the biosynthetic pathway of cephamycin C production in conditions of dissolved oxygen limitation. Some of these compounds, like penicillin N, present antibacterial activity that may interfere in the determination of the cephamycin C concentration by biological methods.…”

Section: Resultsmentioning

confidence: 99%

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An Alternative Methodology for Determination of Cephamycin C from Fermentation Broth

Neto¹

2012

J Chromat Separation Techniq

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Cephamycin C is a β-lactam antibiotic used as a raw material in several commercial antibiotics. The production and purification process of this antibiotic requires a fast, simple and accurate method to quantify it. In this paper, it was developed a high-performance liquid chromatography method to determine cephamycin C concentration, in order to offer an option for the traditional, but laborious, bioassay method normally employed. A method to obtain a calibration curve using the bioassay and cephalosporin C as standard was also proposed. The method showed more efficiency in determining cephamycin C than the bioassay, and it was simpler and faster to execute.

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“…To obtain this antibiotic at an adequate purity level, the use of appropriate downstream techniques is essential to isolate it from the fermentation broth. Separation techniques based on adsorption and ion-exchange processes have been widely applied for the isolation and purification of products obtained during bioprocesses. − These processes stand out from other separation techniques because they provide a high concentration factor, which satisfies the need for a high recovery of the product of interest . Moreover, they are nondenaturing, highly selective, and relatively inexpensive processes .…”

Section: Introductionmentioning

confidence: 99%

Fixed-Bed Column Process as a Strategy for Separation and Purification of Cephamycin C from Fermented Broth

Oliveira

Brites

Bustamante

et al. 2015

Ind. Eng. Chem. Res.

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Fixed-bed column processes using the anionic resin Q Sepharose XL were evaluated for cephamycin C (CepC) purification from fermentation broth. Breakthrough and desorption curves were obtained for different flow rates (2.5, 5.0, and 7.5 mL/min). The elution method consisted of a stepwise gradient using NaCl solutions (0.1%, 0.3%, and 0.5%), which resulted in the separation of CepC from other antibiotics. The flow rate did not interfere in adsorption during loading of the column, but band broadening was observed during elution as the flow rate was increased. After the ion-exchange process, the fractions containing CepC were subjected to solid-phase extraction using a C18 cartridge to remove salts. Analyses of the broth used to feed the column and of the salt-free fractions by LC−MS showed a reduction in the concentration of some contaminants (possibly penicillin N, deacetylcephalosporin C, and deacetoxycephalosporin C) compared to the concentration of CepC. In conclusion, ion exchange followed by adsorption on a C18 adsorbent was demonstrated to be a selective and efficient procedure for the purification of CepC from fermentation broth.

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Preliminary Studies for Cephamycin C Purification Technique

Cited by 6 publications

References 13 publications

Enhancing effect of lysine combined with other compounds on cephamycin C production in Streptomyces clavuligerus

Enhancing effect of lysine combined with other compounds on cephamycin C production in Streptomyces clavuligerus

An Alternative Methodology for Determination of Cephamycin C from Fermentation Broth

Fixed-Bed Column Process as a Strategy for Separation and Purification of Cephamycin C from Fermented Broth

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