Platensimycin (PTM) and platencin (PTN) are potent and selective inhibitors of bacterial and mammalian fatty acid synthases, emerging as promising drug leads for antibacterial and antidiabetic therapies, but suffer from poor pharmacokinetic properties. Previous efforts to prepare analogues by organic synthesis or isolate congeners by fermentation optimization have not significantly improved therapeutic properties. Inspired by the finding that the PTN-overproducing ΔptnR1 mutant strain Streptomyces platensis SB12600 provides an excellent opportunity to isolate PTN congeners that would be produced in titres too low to be detected in the wild-type strain, we subjected S. platensis SB12600 to large-scale fermentation. The crude extract made from approximately 15 l of SB12600 fermentation was subjected to multiple steps of SiO2, C-18 and Sephadex LH-20 chromatography, affording four new PTN congeners (3–6). Structural elucidation of 3–6 as glucoside congeners of PTN was based mainly on HR-ESI-MS and 1H and 13C NMR spectroscopic analyses, as well as comparison with previously reported glucoside congeners of PTM (7 and 8) and PTN (9 and 10). 3 was assigned as platencin A12 (a platencin-5'-β-D-glucoside), 4 as platencin A13 (a 6,7-dihydroplatencin-5'-β-D-glucoside), 5 as platencin A14 (a platencin A1-5'-β-D-glucoside), and 6 as platencin A15 (a platencin A3-5'-β-D-glucoside). The newly isolated four PTN glucosides (3–6) also showed no antibacterial activity as 7–10 reported previously, further supporting the emerging structure–activity relationship that the C-5' hydroxyl group is critical. This work demonstrated once again the power and utility of engineering overproducers by manipulating pathway regulation to isolate natural product congeners that would be produced in titres too low to be detected in the wild-type strains.