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 suffering from poor pharmacokinetic properties. Previous studies engineered Streptomyces platensis SB12600 (a ΔptnR1 mutant strain that overproduces PTN) to facilitate isolation of minor congeners. In this work, S. platensis SB12600 was subjected to large-scale fermentation, and its crude extract was purified via multiple chromatographic steps (SiO₂, C-18, Sephadex LH-20), yielding four new PTN glucoside congeners (3–6). Structural elucidation based on HR-ESI-MS and ¹H/¹³C NMR analyses (including COSY, HMBC, TOCSY, NOESY correlations) and comparison with known analogs identified them as platencin A12 (3, platencin-5'-β-D-glucoside), platencin A13 (4, 6,7-dihydroplatencin-5'-β-D-glucoside), platencin A14 (5, platencin A1-5'-β-D-glucoside), and platencin A15 (6, platencin A3-5'-β-D-glucoside). Similar to previously isolated PTM/PTN glucosides, these new congeners showed no antibacterial activity, further confirming that the C-5' hydroxyl group is critical for FabF/H inhibitory and antibacterial activities. This study highlights the utility of engineering pathway regulation in overproducer strains to isolate low-titre natural product congeners.