Naphthoquinone macrolides have attracted considerable synthetic interest due to their unique scaffolds, but there was no completion of total synthesis reported. In this study, genome mining and genetic manipulations were applied to stimulate the production of naphthoquinone macrolides from the deepsea actinomycete Streptomyces olivaceus SCSIO T05; as a result, new metabolites olimycins A and B (1, 2) were isolated. The structures were elucidated on the basis of extensive spectroscopic data and X-ray single crystal diffraction studies. The results of these efforts enabled rectification of a key structural error made during previous efforts to characterize naphthoquinone macrolides.© 2017 Published by Elsevier Ltd.