scholarly journals Biochemical and Genetic Characterization of Carbapenem-Hydrolyzing β-Lactamase OXA-229 from Acinetobacter bereziniae

2012 ◽  
Vol 56 (7) ◽  
pp. 3923-3927 ◽  
Author(s):  
Rémy A. Bonnin ◽  
Alain A. Ocampo-Sosa ◽  
Laurent Poirel ◽  
Hélène Guet-Revillet ◽  
Patrice Nordmann
Keyword(s):  
Insertion Sequence ◽  
Carbapenem Resistance ◽  
Amino Acid Identity ◽  
Cloning And Expression ◽  
Skin Sample ◽  
Content Type ◽  
Promoter Sequences ◽  
Class D ◽  
Genetic Context

ABSTRACTAcinetobacter bereziniae(formerlyAcinetobactergenomospecies 10) isolate Nec was recovered from a skin sample of a patient hospitalized in Paris, France. It was resistant to penicillins, penicillin-inhibitor combinations, and carbapenems. Cloning and expression inEscherichia coliidentified the carbapenem-hydrolyzing class D β-lactamase OXA-229, which is weakly related to other oxacillinases (66% amino acid identity with the closest oxacillinase, OXA-58). It hydrolyzed penicillins, oxacillin, and imipenem but not expanded-spectrum cephalosporins. Sequencing of the genetic context of theblaOXA-229gene did not identify an insertion sequence but did identify mutations in the promoter sequences in comparison to the fully susceptibleA. bereziniaereference strain. The overexpression ofblaOXA-229inA. bereziniaeNec as a source of carbapenem resistance was identified by quantitative real-time PCR.

scholarly journals First Characterization of CTX-M-15-Producing Escherichia coli Strains Belonging to Sequence Type (ST) 410, ST224, and ST1284 from Commercial Swine in South America

2016 ◽  
Vol 60 (4) ◽  
pp. 2505-2508 ◽  
Author(s):  
Ketrin C. Silva ◽  
Marina Moreno ◽  
Carlos Cabrera ◽  
Beny Spira ◽  
Louise Cerdeira ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
South America ◽  
Insertion Sequence ◽  
Sequence Type ◽  
Content Type ◽  
First Time ◽  
Genetic Context ◽  
New Reservoir

ABSTRACTWe report for the first time the isolation of CTX-M-15-producingEscherichia colistrains belonging to sequence type (ST) 410, ST224, and ST1284 in commercial swine in Brazil. TheblaCTX-M-15gene was located on F-::A9::B1 and C1::A9::B1 IncF-type plasmids, surrounded by a new genetic context comprising the IS26insertion sequence truncated with the ISEcp1element upstream ofblaCTX-M-15. These results reveal that commercial swine have become a new reservoir of CTX-M-15-producing bacteria in South America.

scholarly journals OXA-198, an Acquired Carbapenem-Hydrolyzing Class D β-Lactamase from Pseudomonas aeruginosa

2011 ◽  
Vol 55 (10) ◽  
pp. 4828-4833 ◽  
Author(s):  
Farid El Garch ◽  
Pierre Bogaerts ◽  
Carine Bebrone ◽  
Moreno Galleni ◽  
Youri Glupczynski
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Carbapenem Resistance ◽  
Amino Acid Identity ◽  
Ventilator Associated Pneumonia ◽  
Class 1 Integron ◽  
Content Type ◽  
New Class ◽  
Carbapenem Resistant ◽  
Class D ◽  
Class 1

ABSTRACTA carbapenem-resistantPseudomonas aeruginosastrain (PA41437) susceptible to expanded-spectrum cephalosporins was recovered from several consecutive lower-respiratory-tract specimens of a patient who developed a ventilator-associated pneumonia while hospitalized in an intensive care unit. Cloning experiments identified OXA-198, a new class D β-lactamase which was weakly related (less than 45% amino acid identity) to other class D β-lactamases. Expression inEscherichia coliTOP10 and inP. aeruginosaPAO1 led to transformants that were resistant to ticarcillin and showed reduced susceptibility to carbapenems and cefepime. TheblaOXA-198gene was harbored by a class 1 integron carried by a ca. 46-kb nontypeable plasmid. This study describes a novel class D β-lactamase involved in carbapenem resistance inP. aeruginosa.

scholarly journals OXA-58, a Novel Class D β-Lactamase Involved in Resistance to Carbapenems in Acinetobacter baumannii

2005 ◽  
Vol 49 (1) ◽  
pp. 202-208 ◽  
Author(s):  
Laurent Poirel ◽  
Sophie Marqué ◽  
Claire Héritier ◽  
Christine Segonds ◽  
Gérard Chabanon ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Acinetobacter Baumannii ◽  
Reference Strain ◽  
Carbapenem Resistance ◽  
Amino Acid Identity ◽  
Cloning And Expression ◽  
Acid Identity ◽  
Carbapenem Resistant ◽  
Class D ◽  
Insertion Elements

ABSTRACT A carbapenem-resistant Acinetobacter baumannii strain was isolated in Toulouse, France, in 2003. Cloning and expression in Escherichia coli identified the carbapenem-hydrolyzing β-lactamase OXA-58, which is weakly related (less than 50% amino acid identity) to other oxacillinases. It hydrolyzed penicillins, oxacillin, and imipenem but not expanded-spectrum cephalosporins. The bla OXA-58 gene was located on a ca. 30-kb non-self-transferable plasmid. After electrotransformation in the A. baumannii CIP7010T reference strain, it conferred reduced susceptibility to carbapenems. The bla OXA-58 gene was bracketed by two novel ISAba3-like insertion elements. This study describes a newly characterized β-lactamase that may contribute to carbapenem resistance in A. baumannii.

scholarly journals Characterization of OXA-181, a Carbapenem-Hydrolyzing Class D β-Lactamase from Klebsiella pneumoniae

2011 ◽  
Vol 55 (10) ◽  
pp. 4896-4899 ◽  
Author(s):  
Anaïs Potron ◽  
Patrice Nordmann ◽  
Emilie Lafeuille ◽  
Zaina Al Maskari ◽  
Fatma Al Rashdi ◽  
...  
Keyword(s):  
Amino Acid ◽  
Klebsiella Pneumoniae ◽  
Insertion Sequence ◽  
Amino Acid Substitutions ◽  
Sultanate Of Oman ◽  
Content Type ◽  
Class D

ABSTRACTKlebsiella pneumoniaeKP3 was isolated from a patient transferred from India to the Sultanate of Oman.K. pneumoniaeKP3 was resistant to all β-lactams, including carbapenems, and expressed the carbapenem-hydrolyzing β-lactamase OXA-181, which differs from OXA-48 by four amino acid substitutions. Compared to OXA-48, OXA-181 possessed a very similar hydrolytic profile. TheblaOXA-181gene was located on a 7.6-kb ColE-type plasmid and was linked to the insertion sequence ISEcp1. The ISEcp1-mediated one-ended transposition ofblaOXA-181was also demonstrated.

scholarly journals High-Level Carbapenem Resistance in OXA-232-Producing Raoultella ornithinolytica Triggered by Ertapenem Therapy

2019 ◽  
Vol 64 (1) ◽  
Author(s):  
Alina Iovleva ◽  
Roberta T. Mettus ◽  
Christi L. McElheny ◽  
Marissa P. Griffith ◽  
Mustapha M. Mustapha ◽  
...  
Keyword(s):  
Rapid Development ◽  
Carbapenem Resistance ◽  
Resistant Isolate ◽  
Clinical Microbiology Laboratory ◽  
Loss Of Function ◽  
Content Type ◽  
Carbapenem Resistant ◽  
Class D ◽  
Fold Reduction ◽  
High Level

ABSTRACT OXA-232 is an OXA-48-group class D β-lactamase that hydrolyzes expanded-spectrum cephalosporins and carbapenems at low levels. Clinical strains producing OXA-232 are sometimes susceptible to carbapenems, making it difficult to identify them in the clinical microbiology laboratory. We describe the development of carbapenem resistance in sequential clinical isolates of Raoultella ornithinolytica carrying blaOXA-232 in a hospitalized patient, where the ertapenem MIC increased from 0.5 μg/ml to 512 μg/ml and the meropenem MIC increased from 0.125 μg/ml to 32 μg/ml during the course of ertapenem therapy. Whole-genome sequencing (WGS) analysis identified loss-of-function mutations in ompC and ompF in carbapenem-resistant isolates that were not present in the initial carbapenem-susceptible isolate. Complementation of a carbapenem-resistant isolate with an intact ompF gene resulted in 16- to 32-fold reductions in carbapenem MICs, whereas complementation with intact ompC resulted in a 2-fold reduction in carbapenem MICs. Additionally, blaOXA-232 expression increased 2.9-fold in a carbapenem-resistant isolate. Rapid development of high-level carbapenem resistance in initially carbapenem-susceptible OXA-232-producing R. ornithinolytica under selective pressure from carbapenem therapy highlights the diagnostic challenges in detecting Enterobacteriaceae strains producing this inefficient carbapenemase.

scholarly journals Horizontal Transfer of the OXA-24 Carbapenemase Gene via Outer Membrane Vesicles: a New Mechanism of Dissemination of Carbapenem Resistance Genes in Acinetobacter baumannii

2011 ◽  
Vol 55 (7) ◽  
pp. 3084-3090 ◽  
Author(s):  
Carlos Rumbo ◽  
Esteban Fernández-Moreira ◽  
María Merino ◽  
Margarita Poza ◽  
Jose Antonio Mendez ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Outer Membrane ◽  
Resistance Genes ◽  
Carbapenem Resistance ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Content Type ◽  
Class D ◽  
Carbapenemase Gene

ABSTRACTThe resistance ofAcinetobacter baumanniistrains to carbapenems is a worrying problem in hospital settings. The main mechanism of carbapenem resistance is the expression of β-lactamases (metalloenzymes or class D enzymes). The mechanisms of the dissemination of these genes amongA. baumanniistrains are not fully understood. In this study we used two carbapenem-resistant clinical strains ofA. baumannii(AbH12O-A2 and AbH12O-CU3) expressing the plasmid-borneblaOXA-24gene (plasmids pMMA2 and pMMCU3, respectively) to demonstrate thatA. baumanniireleases outer membrane vesicles (OMVs) duringin vitrogrowth. The use of hybridization studies enabled us to show that these OMVs harbored theblaOXA-24gene. The incubation of these OMVs with the carbapenem-susceptibleA. baumanniiATCC 17978 host strain yielded full resistance to carbapenems. The presence of the original plasmids harboring theblaOXA-24gene was detected in strain ATCC 17978 after the transformation of OMVs. New OMVs harboringblaOXA-24were released byA. baumanniiATCC 17978 after it was transformed with the original OMV-mediated plasmids, indicating the universality of the process. We present the first experimental evidence that clinical isolates ofA. baumanniimay release OMVs as a mechanism of horizontal gene transfer whereby carbapenem resistance genes are delivered to surroundingA. baumanniibacterial isolates.

scholarly journals Characterization of BRPMBL, the Bleomycin Resistance Protein Associated with the Carbapenemase NDM

2017 ◽  
Vol 61 (3) ◽  
Author(s):  
Laurent Dortet ◽  
Delphine Girlich ◽  
Anne-Laure Virlouvet ◽  
Laurent Poirel ◽  
Patrice Nordmann ◽  
...  
Keyword(s):  
Specific Activity ◽  
Cloning And Expression ◽  
Cross Resistance ◽  
Dna Protection ◽  
Mobility Shift ◽  
Content Type ◽  
Reverse Transcription Pcr ◽  
Resistance Protein

ABSTRACT The metallo-β-lactamase NDM-1 is among the most worrisome resistance determinants and is spreading worldwide among Gram-negative bacilli. A bleomycin resistance gene, ble MBL, downstream of the bla NDM-1 gene has been associated with resistance almost systematically. Here, we characterized the corresponding protein, BRPMBL, conferring resistance to bleomycin, an antitumoral glycopeptide molecule. We have determined whether the expression of the bla NDM-1-ble MBL operon is inducible in the presence of carbapenems and/or bleomycin-like molecules using quantitative reverse transcription-PCR (qRT-PCR), determination of imipenem and zeocin MICs, and carbapenemase-specific activity assays. We showed that the bla NDM-1-ble MBL operon is constitutively expressed. Using electrophoretic mobility shift and DNA protection assays performed with purified glutathione S-transferase (GST)-BRPMBL, we demonstrated that BRPMBL is able to bind and sequester bleomycin-like molecules, thus preventing bleomycin-dependent DNA degradation. In silico modeling confirmed that the mechanism of action required the dimerization of the BRPMBL protein in order to sequester bleomycin and prevent DNA damage. BRPMBL acts specifically on bleomycin-like molecules since cloning and expression of ble MBL in Staphyloccoccus aureus did not confer cross-resistance to any other antimicrobial glycopeptides such as vancomycin and teicoplanin.

scholarly journals Genetic and Biochemical Characterization of an Acquired Subgroup B3 Metallo-β-Lactamase Gene,blaAIM-1, and Its Unique Genetic Context in Pseudomonas aeruginosa from Australia

2012 ◽  
Vol 56 (12) ◽  
pp. 6154-6159 ◽  
Author(s):  
Dongeun Yong ◽  
Mark A. Toleman ◽  
Jan Bell ◽  
Brett Ritchie ◽  
Rachael Pratt ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Southern Hybridization ◽  
Biochemical Characterization ◽  
Gene Bank ◽  
Clinical Isolates ◽  
Gram Negative ◽  
Content Type ◽  
Lactamase Gene ◽  
Genetic Context

ABSTRACTThree clinicalPseudomonas aeruginosaisolates (WCH2677, WCH2813, and WCH2837) isolated from the Women's and Children's Hospital, Adelaide, Australia, produced a metallo-β-lactamase (MBL)-positive Etest result. All isolates were PCR negative for known MBL genes. A gene bank was created, and an MBL gene, designatedblaAIM-1, was cloned and fully characterized. The encoded enzyme, AIM-1, is a group B3 MBL that has the highest level of identity to THIN-B and L1. It is chromosomal and flanked by two copies (one intact and one truncated) of an ISCRelement, ISCR15. Southern hybridization studies indicated the movement of both ISCR15andblaAIM-1within the three different clinical isolates. AIM-1 hydrolyzes most β-lactams, with the exception of aztreonam and, to a lesser extent, ceftazidime; however, it possesses significantly higherkcatvalues for cefepime and carbapenems than most other MBLs. AIM-1 was the first mobile group B3 enzyme detected and signals further problems for already beleaguered antimicrobial regimes to treat seriousP. aeruginosaand other Gram-negative infections.

scholarly journals 4-Amino-2-Sulfanylbenzoic Acid as a Potent Subclass B3 Metallo-β-Lactamase-Specific Inhibitor Applicable for Distinguishing Metallo-β-Lactamase Subclasses

2019 ◽  
Vol 63 (10) ◽  
Author(s):  
Jun-ichi Wachino ◽  
Reo Kanechi ◽  
Erina Nishino ◽  
Marie Mochizuki ◽  
Wanchun Jin ◽  
...  
Keyword(s):  
Specific Inhibitor ◽  
Carbapenem Resistance ◽  
Resistance Mechanisms ◽  
Health Concern ◽  
Conserved Residues ◽  
Content Type ◽  
Carbapenem Resistant ◽  
Promising Strategy ◽  
Starting Point

ABSTRACT The number of cases of infection with carbapenem-resistant Enterobacteriaceae (CRE) has been increasing and has become a major clinical and public health concern. Production of metallo-β-lactamases (MBLs) is one of the principal carbapenem resistance mechanisms in CRE. Therefore, developing MBL inhibitors is a promising strategy to overcome the problems of carbapenem resistance conferred by MBLs. To date, the development and evaluation of MBL inhibitors have focused on subclass B1 MBLs but not on B3 MBLs. In the present study, we searched for B3 MBL (specifically, SMB-1) inhibitors and found thiosalicylic acid (TSA) to be a potent inhibitor of B3 SMB-1 MBL (50% inhibitory concentration [IC50], 0.95 μM). TSA inhibited the purified SMB-1 to a considerable degree but was not active against Escherichia coli cells producing SMB-1, as the meropenem (MEM) MIC for the SMB-1 producer was only slightly reduced with TSA. We then introduced a primary amine to TSA and synthesized 4-amino-2-sulfanylbenzoic acid (ASB), which substantially reduced the MEM MICs for SMB-1 producers. X-ray crystallographic analyses revealed that ASB binds to the two zinc ions, Ser221, and Thr223 at the active site of SMB-1. These are ubiquitously conserved residues across clinically relevant B3 MBLs. ASB also significantly inhibited other B3 MBLs, including AIM-1, LMB-1, and L1. Therefore, the characterization of ASB provides a starting point for the development of optimum B3 MBL inhibitors.

scholarly journals Molecular Characterization of OXA-20, a Novel Class D β-Lactamase, and Its Integron from Pseudomonas aeruginosa

1998 ◽  
Vol 42 (8) ◽  
pp. 2074-2083 ◽  
Author(s):  
Thierry Naas ◽  
Wladimir Sougakoff ◽  
Anne Casetta ◽  
Patrice Nordmann
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Amino Acid ◽  
Clavulanic Acid ◽  
Antibiotic Resistance Gene ◽  
Amino Acid Identity ◽  
Acid Identity ◽  
Class D ◽  
Acid Protein ◽  
Amino Acid Protein

ABSTRACT The Pseudomonas aeruginosa Mus clinical isolate produces OXA-18, a pI 5.5 class D extended-spectrum β-lactamase totally inhibited by clavulanic acid (L. N. Philippon, T. Naas, A.-T. Bouthors, V. Barakett, and P. Nordmann, Antimicrob. Agents Chemother. 41:2188–2195, 1997). A second β-lactamase was cloned, and the recombinant Escherichia coli clone pPL10 expressed a pI 7.4 β-lactamase which conferred high levels of amoxicillin and ticarcillin resistance and which was partially inhibited by clavulanic acid. The 2.5-kb insert from pPL10 was sequenced, and a 266-amino-acid protein (OXA-20) was deduced; this protein has low amino acid identity with most of the class D β-lactamases except OXA-2, OXA-15, and OXA-3 (75% amino acid identity with each). OXA-20 is a restricted-spectrum oxacillinase and is unusually inhibited by clavulanic acid. OXA-20 is a peculiar β-lactamase because its translation initiates with a TTG (leucine) codon, which is rarely used as a translational origin in bacteria. Exploration of the genetic environment of oxa20revealed the presence of the following integron features: (i) a second antibiotic resistance gene, aacA4; (ii) anintI1 gene; and (iii) two 59-base elements, each associated with either oxa20 or aacA4. This integron is peculiar because it lacks the 3′ conserved region, and therefore is not a sul1-associated integron like most of them, and because its 3′ end is located within tnpR, a gene involved in the transposition of Tn5393, a gram-negative transposon.P. aeruginosa Mus produces two novel and unrelated oxacillinases, OXA-18 and OXA-20, both of which are inhibited by clavulanic acid.

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