Large crystals of arylesterase from Pseudomonas fluorescens have been grown at room temperature using ammonium sulfate as a precipitant. They grow to dimensions of 0.7 x 0.7 x 0.6 mm3 within a month. The crystals belong to the trigonal space group P3(1) (or P3(2)), with unit cell dimensions of a = 147.12 angstrom and c = 131.08 angstrom. The asymmetric unit seems to contain six molecules of dimeric arylesterase, with corresponding crystal volume per protein mass (V(M)) of 2.53 angstrom3/Da and solvent fraction of 51.5% by volume. The crystals diffract to at least 2.2 angstrom Bragg spacing when exposed to X-rays from a rotating-anode source. X-ray data have been collected to 2.9 angstrom Bragg spacing from native crystals.