title: Chromosomal beta-lactamase of Klebsiella oxytoca, a new class A enzyme that hydrolyzes broad-spectrum beta-lactam antibiotics.
abstract: The chromosomally encoded beta-lactamase gene of Klebsiella oxytoca E23004, a strain resistant to cefoperazone and aztreonam, was cloned and expressed in Escherichia coli HB101. The molecular mass and pI of this enzyme were 28 kilodaltons and 7.4, respectively. Although the beta-lactamase of K. oxytoca hydrolyzed many cephalosporins, including broad-spectrum drugs, the nucleotide sequence and deduced amino acid sequence lacked homology with chromosomal class C beta-lactamase genes (ampC) of E. coli or Citrobacter freundii. Rather, about 45% nucleotide sequence homology and 40% deduced amino acid sequence homology were observed between the K. oxytoca beta-lactamase and TEM-1, a class A beta-lactamase which does not efficiently hydrolyze cephalosporins. Values of Km, relative Vmax, and relative Vmax/Km for the K. oxytoca beta-lactamase indicated that the enzyme is a penicillinase but that it can hydrolyze cefoperazone effectively and other broad-spectrum cephems weakly. Hence, the chromosomal beta-lactamase of K. oxytoca E23004 belongs to class A but differences in its amino acid sequence provide a broader spectrum of activity.
authors.affiliation: Department of Bacteriology, Nagoya University School of Medicine, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Arakawa
authors.forename: M
authors.initials: M
authors.lastname: Ohta
authors.forename: N
authors.initials: N
authors.lastname: Kido
authors.forename: M
authors.initials: M
authors.lastname: Mori
authors.forename: H
authors.initials: H
authors.lastname: Ito
authors.forename: T
authors.initials: T
authors.lastname: Komatsu
authors.forename: Y
authors.initials: Y
authors.lastname: Fujii
authors.forename: N
authors.initials: N
authors.lastname: Kato
mesh_terms: D000667:Ampicillin / Q000494:pharmacology; D001483:Base Sequence; D002439:Cefotaxime / Q000494:pharmacology; D002509:Cephaloridine / Q000494:pharmacology; D002876:Chromosomes, Bacterial / Q000378:metabolism; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000032:analysis*; D004352:Drug Resistance, Microbial / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D007709:Klebsiella / Q000201:enzymology* / Q000235:genetics; D007711:Klebsiella pneumoniae / Q000201:enzymology / Q000235:genetics; D015183:Restriction Mapping; D001618:beta-Lactamases / Q000032:analysis / Q000235:genetics* / Q000378:metabolism
chemical_list: D004269:DNA, Bacterial; D000667:Ampicillin; D001618:beta-Lactamases; D002509:Cephaloridine; D002439:Cefotaxime