DNA fragments encoding two putative zinc-dependent hydrolases, designated GLX2-1 and GLX2-2, from a clinical isolate of Campylobacter jejuni, strain 012, were cloned and sequenced. GLX2-1 was encoded by a sequence of 798 bp and GLX2-2 by a sequence of 597 bp. The amino acid sequences deduced from C. jejuni DNA showed 99% and 100% identity, respectively, to putative zinc hydrolases reported from C. jejuni ATCC strain 11168, and also shared identity (28-43%) with several hypothetical conserved proteins and known zinc-dependent hydrolases and metallo-beta-lactamase superfamily proteins. A strictly conserved motif, -H-X-H-X-D-, characteristic of the metallo-beta-lactamase superfamily of proteins, including class B metallo-beta-lactamases, was identified in both proteins. Other conserved metal-binding ligands, characteristic of the metallo-beta-lactamase superfamily of proteins, were also identified. Functional beta-lactamase could not be expressed in either Escherichia coli or Campylobacter coli transformed with C. jejuni hydrolase-containing plasmids, suggesting that they do not function as metallo-beta-lactamases, although structurally they are consistent with the zinc metallo-hydrolase family of the beta-lactamase fold.