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Notonesomycin A is a 32-membered bioactive glycosylated macrolactone known to be produced by  Streptomyces aminophilus  subsp.  notonesogenes  647-AV1 and  S. aminophilus  DSM 40186. In a high throughput antifungal screening campaign, we identified an alternative notonesomycin A producing strain,  Streptomyces  sp. A793, and its biosynthetic gene cluster. From this strain, we further characterized a new more potent antifungal non-sulfated analogue, named notonesomycin B. Through CRISPR–Cas9 engineering of the biosynthetic gene cluster, we were able to increase the production yield of notonesomycin B by up to 18-fold as well as generate a strain that exclusively produces this analogue.

Identification and engineering of 32 membered antifungal macrolactone notonesomycins