Actinomycetes have been a source of anti-infectives for more than 50 years and continue to play an important role in new antibiotic discovery. In a screening program for new antibacterial compounds, a Streptomyces strain (Lv20-58) isolated from the root zone of Oleaceae europea was investigated. The strain was cultivated in M medium, extracted with ethyl acetate, and purified via semipreparative reverse-phase HPLC to yield two new cyclic hexadepsipeptides, oleamycins A and B. Structural analysis using HR-ESI(+)MS and NMR revealed that oleamycin A (molecular formula C39H66N8O11) is a 19-membered cyclic hexapeptide consisting of a 3-hydroxyleucine residue, three glycine residues (two of which are N-methylated), two piperazic acid residues, and a polyketide side chain. Oleamycin B (molecular formula C38H64N8O11) has a similar structure with a slightly different molecular formula. Both compounds are new antibacterial cyclic hexadepsipeptides isolated from a terrestrial Streptomyces sp.