Engineering the production of a conformational variant of occidiofungin that has enhanced inhibitory activity against fungal species

摘要

Occidiofungin is a cyclic nonribosomally synthesized antifungal peptide with submicromolar activity. This invention is directed to compositions enriched for particular occidiofungin diastereomers/conformers, methods of making compositions enriched for particular diastereomers/conformers and microorganisms suitable for producing enriched compositions of particular diastereomers/conformers. Methods of treating fungal infections or plants infected by fungi are also provided.

主权项

1. A method of increasing production of occidiofungin diastereomers/conformers corresponding to the diastereomers/conformers having the total correlation spectroscopy (TOCSY) fingerprint set forth in FIG. 5C as the green NH correlations in a microorganism producing occidiofungin comprising:
decreasing the thioesterase activity of an occidiofungin gene D (ocfD) product (OcfD) in said microorganism, wherein said OcfD thioesterase activity is decreased by the introduction of a point mutation at the serine in position 2954 of the amino acid sequence of SEQ ID NO: 4, said point mutation replaces said serine with an alanine, glycine or proline, and culturing said microorganism under conditions to produce said occidiofungin diastereomers/conformers, wherein said increase in production of occidiofungin diastereomers/conformers is as compared to a reference microorganism without said decrease in OcfD thioesterase activity cultured under the same conditions.