Abstract
A protocol for preparative isopenicillin N (IPN) purification, a highly interesting and hitherto unavailable intermediate of the penicillin and cephalosporin biosynthetic pathway due to its high unstability, is described. Culture broths of Acremonium chrysogenum TD189, a strain blocked in cephalosporin biosynthesis that accumulates this metabolite, were treated with acetone and filtered though charcoal and a hydrophobic resin in a single step as tandem columns. The cleared broth was then lyophilized and passed though a Sephadex G-25 column. The last step was the purification to homogeneity of IPN in a semipreparative HPLC equipment and, optionally, a desalting step by Sephadex G-10 column. Once purified, a complete analysis of the stability of the compound and the conditions for its long-term storage was carried out. Our results suggest a first-order model for IPN decomposition for all the pH and temperature analyzed. IPN is more stable at neutral pH, and once lyophilized, can be stored under vacuum and −75 °C with a half-life of 770 days.
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Acknowledgements
This work was supported by a grant from the European Union (Eurofungbase, LSSGCT2005018964) to JFM. I Vaca thanks for a fellowship of the Diputación de León (Spain) and the support of PBCT program of CONICYT (Chile), project PDA13. R Chávez thanks to DICYT-USACH for support. We thank C García-Estrada for scientific support and B Martín and J Merino for excellent technical assistance.
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Vaca, I., Casqueiro, J., Ullán, R. et al. A preparative method for the purification of isopenicillin N from genetically blocked Acremonium chrysogenum strain TD189: studies on the degradation kinetics and storage conditions. J Antibiot 64, 447–451 (2011). https://doi.org/10.1038/ja.2011.30
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DOI: https://doi.org/10.1038/ja.2011.30
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