Bioassay-directed fractionation of the butanol extract of Streptomyces sp. L-4-4, using the hyphae formation inhibition assay of a prokaryotic whole cell, led to the isolation of six new aminocoumarins, coumabiocins A-F (1-6), along with two known compounds, novobiocin (7) and isonovobiocin (8). Coumabiocins A-E (1-5) contain three structural elements, a central 3-amino-7-hydroxycoumarin that is linked at the 3-amino group to a prenylated 4-hydroxybenzoic acid moiety and at the 7-position to an L-noviosyl sugar, while coumabiocin F (6) lacks the sugar moiety. Their structures were elucidated by spectroscopic methods including 1D- and 2D-NMR techniques and mass spectrometric analyses. Coumabiocins A-E (1-5) exhibited significant inhibitory activity against Streptomyces 85E and gave a 10-15 mm clear zone of inhibition at 20 µg/disk and a 10 mm bald and a 10 mm clear zone of inhibition at 5 and 10 µg/disk, respectively, whereas coumabiocin F (6) was inactive.