Staphylococcus aureusis the causative agent of many diseases, including meningitis, bacteraemia, pneumonia, food poisoning and toxic shock syndrome. Structural characterization of the PaaI-like thioesterase SAV0944 (SaPaaI) fromS. aureussubsp.aureusMu50 will aid in understanding its potential as a new therapeutic target by knowledge of its molecular details and cellular functions. Here, the recombinant expression, purification and crystallization ofSaPaaI thioesterase fromS. aureusare reported. This protein initially crystallized with the ligand coenzyme A using the hanging-drop vapour-diffusion technique with condition No. 40 of Crystal Screen from Hampton Research at 296 K. Optimal final conditions consisting of 24% PEG 4000, 100 mMsodium citrate pH 6.5, 12% 2-propanol gave single diffraction-quality crystals. These crystals diffracted to beyond 2 Å resolution at the Australian Synchrotron and belonged to space groupP1211, with unit-cell parametersa= 44.05,b= 89.05,c= 60.74 Å, β = 100.5°. Initial structure determination and refinement gave anRfactor andRfreeof 17.3 and 22.0%, respectively, confirming a positive solution in obtaining phases using molecular replacement.