(1S,4R)-(-)-4-Hydroxy-2-cyclopentenyl acetate (2), a versatile intermediate in prostaglandin syntheses, was readily prepared by an efficient enzyme-catalyzed enantioselective monoacetylation of cis-2-cyclopenten-1,4-diol (1) with 2,2,2-trichloroethyl acetate in the organic solvent system triethylamine/tetrahydrofuran. The chemical yield reached nearly 50%. The enantiomeric excess of the crude product was 95%. It could be raised to more than 99% by a single recrystallization. Commercially available pancreatin, a crude enzyme preparation from porcine pancreas, was used as biocatalyst.