A chromosomally encoded class D beta-lactamase, OXA-114, was characterized from Achromobacter xylosoxidans strain CIP69598. beta-Lactamase OXA-114 shared 56% amino acid identity with the naturally occurring class D beta-lactamase of Burkholderia cenocepacia and 42% identity with the acquired oxacillinases OXA-9 and OXA-18. OXA-114 has a narrow-spectrum hydrolysis profile, although it includes imipenem, at a very low level. PCR and sequencing revealed that bla(OXA-114)-like genes were identified in all A. xylosoxidans strains tested (n = 5), indicating that this beta-lactamase is naturally occurring in that species. Induction experiments with imipenem and cefoxitin did not show inducibility of bla(OXA-114) expression.