Acuminolide A (1), along with pectenotoxin II (PTX-2), dinophysistoxin I (DTX-1), okadaic acid (OA), and 7-epi-PTX-2 seco acid, was isolated from a large-scale cultivation of the dinoflagellate Dinophysis acuminata. The new 33-membered macrolide 1 was characterized by detailed analysis of 2D NMR and MS data. Its relative stereochemistry was elucidated on the basis of ROESY correlations and J-based analysis. In contrast to the other well-known toxins that were isolated, 1 showed no cytotoxicity against four cancer cell lines but caused potent stimulation of actomyosin ATPase activity.