title: Resistance to oxyimino beta-lactams due to a mutation of chromosomal beta-lactamase in Citrobacter freundii.
abstract: The duplicative mutation of an Ala-Val-Arg sequence at positions 208 to 210 in the loop structure of Enterobacter cloacae class C beta-lactamase caused substrate specificity extension to oxyimino beta-lactam antibiotics and this chromosomal mutation provided bacterial cells with high resistance to the beta-lactams (M. Nukaga et al, 1995, J. Biol. Chem. 270, 5729-5735). In order to confirm the universality of this phenomenon among other class C beta-lactamases, the duplicative mutation was applied to a class C beta-lactamase of Citrobacter freundii, which has 74% homology to the E. cloacae beta-lactamase amino acid sequence. The counterpart sequence to the Ala-Val-Arg of the E. cloacae enzyme in C. freundii beta-lactamase was identified to be Pro-Val-His. A Pro-Val-His sequence was inserted just after the native Pro-Val-His sequence at positions 208 to 210 in the C. freundii beta-lactamase. The resulting mutant of C. freundii beta-lactamase obtained a striking characteristic that we expected, showing substrate specificity extension to oxyimino beta-lactams. Nearly the same result was obtained with the insertion of an Ala-Val-Arg sequence after the native Pro-Val-His sequence. These results indicate that structural modification of this locus commonly induces modification of the substrate specificity to unfavorable substrates for many chromosomal class C beta-lactamases produced by gram-negative bacteria.
authors.affiliation: Division of Microbial Chemistry, Faculty of Pharmaceutical Sciences, Chiba University, Chiba, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Haruta
authors.forename: M
authors.initials: M
authors.lastname: Nukaga
authors.forename: K
authors.initials: K
authors.lastname: Taniguchi
authors.forename: T
authors.initials: T
authors.lastname: Sawai
mesh_terms: D000595:Amino Acid Sequence; D018550:Cephalosporin Resistance / Q000235:genetics; D016971:Citrobacter freundii / Q000235:genetics*; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010403:Penicillin Resistance / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D018440:beta-Lactam Resistance / Q000235:genetics*; D001618:beta-Lactamases / Q000145:classification / Q000235:genetics* / Q000378:metabolism
chemical_list: D001618:beta-Lactamases