Purification and characterization of inducible beta-lactamases in Aeromonas spp.
beta-Lactamases from Aeromonas hydrophila and A. sobria were purified and characterized. Both species produced beta-lactamases that were inducible by either cefoxitin or imipenem. These species were resistant to ampicillin and cephalothin but not imipenem. Isoelectric focusing of sonic extracts revealed one band at pI 8.0 and a second band at pI 7.0 for A. hydrophila. Likewise, A. sobria produced two bands, one at pI 8.4 and the other at pI 7.0. Two enzymes from each species were separated by flatbed electrofocusing gel and purified to homogeneity. The molecular weight of the pI 7.0 enzyme (A1) from both species was estimated to be 42,500, whereas the pI 8.0 (A2h) and 8.4 (A2s) enzymes of A. hydrophila and A. sobria had molecular weights of 31,500 and 35,000, respectively, on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The relative Vmax values for cephalothin, penicillin, and imipenem for these enzymes indicated that A1 was primarily a cephalosporinase while A2h and A2s were penicillinases highly active against carbapenems. A1 was susceptible to inhibition by cloxacillin, while the A2 enzymes were inhibited by clavulanic acid and EDTA and required zinc for activity. Thus, there appear to be two distinct inducible beta-lactamases in A. hydrophila and A. sobria that play an important role in the beta-lactam resistance of these species.
Iaconis
J P
JP
Department of Medical Microbiology, Creighton University School of Medicine, Omaha, Nebraska 68178.
Sanders
C C
CC
D000333:Aeromonas / Q000187:drug effects / Q000201:enzymology*; D000900:Anti-Bacterial Agents / Q000378:metabolism / Q000494:pharmacology; D002969:Clavulanic Acids / Q000494:pharmacology; D003023:Cloxacillin / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004790:Enzyme Induction / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D013056:Spectrophotometry, Ultraviolet; D001618:beta-Lactamases / Q000032:analysis / Q000096:biosynthesis / Q000302:isolation & purification*
D000900:Anti-Bacterial Agents; D002969:Clavulanic Acids; D001618:beta-Lactamases; D003023:Cloxacillin