We measured the kinetics of hydrolysis of various cephalosporins by the chromosomally-encoded beta-lactamases of Citrobacter diversus ULA-27. Cefonicid, cefamandole, cefatrizine and cefoperazone were all hydrolyzed but these antibiotics showed a different feature in their kinetic parameters. Moreover, cefoperazone was a non-competitive inhibitor of this type of enzyme. Cefotetan was stable to hydrolysis and behaved like a progressive inactivator. The ability of these enzymes to inactivate the reported antibiotics contributes largely to the resistance of the studied strain. We conclude that hydrolysis is the main mechanism of resistance of this strain to the new cephalosporins.