期刊
ANTIMICROBIAL AGENTS AND CHEMOTHERAPY
卷 44, 期 6, 页码 1470-1478出版社
AMER SOC MICROBIOLOGY
DOI: 10.1128/AAC.44.6.1470-1478.2000
关键词
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A naturally occurring AmpC beta-lactamase (cephalosporinase) gene was cloned from the Hafnia alvei 1 clinical isolate and expressed in Escherichia coli, The deduced AmpC beta-lactamase (ACC-2) had a pI of 8 and a relative molecular mass of 37 kDa and showed 50 and 47% amino acid identity with the chromosome-encoded AmpCs from Serratia marcescens and Providentia stuartii, respectively. It had 94% amino acid identity with the recently described plasmid-borne cephalosporinase ACC-1 from Klebsiella pneumoniae, suggesting the chromosomal origin of ACC-1, The hydrolysis constants (k(cat) and K-m) showed that ACC-2 was a peculiar cephalosporinase. since it significantly hydrolyzed cefpirome, Once its gene was cloned and expressed in E, coli (pDEL-1), ACC-2 conferred resistance to ceftazidime and cefotaxime but also an uncommon reduced susceptibility to cefpirome, A divergently transcribed ampR gene with an overlapping promoter compared with ampC (bla(ACC-2)) was identified in H. alvei 1, encoding an AmpR protein that shared 64% amino acid identity with the closest AmpR protein from P. stuartii, beta-Lactamase induction experiments showed that the ampC gene was repressed in the absence of ampR and was activated when cefoxitin or imipenem was added as an inducer. From H, alvei 1 cultures that expressed an inducible-cephalosporinase phenotype, several ceftazidime- and cefpirome-cross-resistant H. alvei 1 mutants were obtained upon selection on cefpirome- or ceftazidime-containing plates, and H. alvei 1 DER, a ceftazidime-resistant mutant, stably overproduced cephalosporinase, Transformation of H. alvei 1 DER or E. coli JRG582 (ampDE mutant) harboring ampC and ampR from H. alt ei 1 with a recombinant plasmid containing ampD from E. coli resulted in a decrease in the MIC of beta-lactam and recovery of an inducible phenotype for H. alvei 1 DER Thus, AmpR and AmpD proteins mag regulate biosynthesis of the H, alvei cephalosporinase similarly to other enterobacterial cephalosporinases.
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