Ostalactones A-C (1-3), three new β- and ε-lactone natural products, were isolated from the culture broth of the basidiomycete Stereum ostrea. The structures were elucidated by interpretation of HRFABMS and 1D and 2D NMR data. The structures of 1 and 2 are characterized by the presence of a β-lactone containing a fused 4/5 bicyclic core structure. Compound 3 possesses a 2-oxepinone ring system, which is likely to be a biosynthetic precursor of compounds 1 and 2. Ostalactones A (1) and B (2) displayed potent inhibitory activity against human pancreatic lipase.