Inactivation of cephamycins by various beta-lactamases from gram-negative bacteria

The enzymic inactivation of cephamycins, i.e. cefoxitin, cefmetazole, cefotetan and cefbuperazone, was investigated by means of bioassay, high pressure liquid chromatography (HPLC) and spectrophotometric analysis using three types of cephalosporinase (CSase, RICHMOND type Ia, Ib and Ic) and one penicillinase (PCase, TEM type). These four cephamycins were not inactivated by Ic CSase and TEM type PCase or producers of these enzymes. However, the inactivation of cefmetazole and cefoxitin was noted when they were incubated in the cultures of CSase (Ia and Ib)-producers or incubated with a large amount of these purified enzymes although the inactivation of cefbuperazone was not noted. HPLC of culture fluid or enzyme solution which contained cefmetazole or cefoxitin and were incubated at 37 degrees C showed that metabolites of cefmetazole or cefoxitin appeared as the drug disappeared. In addition, the appearance of metabolites corresponded to the loss of the drug's bioactivities and the absorption of iodine. UV and IR spectra of cefmetazole which were taken after incubation with the purified CSase showed the cleavage of the beta-lactam ring.