Production, purification, and mass spectrometry characterization of the cyclohexadepsipeptide enniatin J₃ and study of the cytoxicity on differentiated and undifferentiated Caco-2 cells

Abstract

The enniatins (EN) are bioactive compounds of hexadepsipeptidic structure produced by several strains of Fusarium spp. The enniatin J₃ (EN J₃) was purified from extracts of Fusarium solani growth on solid medium of kamut, using semipreparative liquid chromatography (LC) followed by an analytical LC. The purity and the structure of the isolated compound were confirmed by electrospray ionization-mass spectrometry study-linear ion trap (ESI-MS-LIT). The pure fraction of EN J₃ was utilized to study the cytotoxicity on differentiated and undifferentiated Caco-2 cells. The use of both chromatographic techniques allowed us to produce and purify 50 mg of EN J₃ completely characterized with the technique of the ESI-MS-LIT. No statistically significant differences over control were detected on Caco-2 cells exposed to EN J₃ at any of the concentrations used.

Keywords:

• enniatin J₃
• LC
• ESI-MS-LIT
• Caco-2 cells
• MTT assay

Acknowledgments

This study was supported by the Science and Education Spanish Ministry Projects (CTQ2007-291 63186 and AGL2007-61493), and by the pre PhD program of University of Valencia “Cinc segles.”