Chromosomal beta-lactamase of Klebsiella oxytoca, a new class A enzyme that hydrolyzes broad-spectrum beta-lactam antibiotics.

QPX7728 is a cyclic boronate ultra-broad-spectrum beta-lactamase inhibitor, with potent activity against both serine and metallo beta-lactamases. QPX7728 can be delivered systemically by the IV or oral route of administration. Oral β-lactam antibiotics alone or in combination with QPX7728 were evaluated for 1) sensitivity to hydrolysis by various common beta-lactamases and inhibition of hydrolysis by QPX7728; 2) the impact of non-beta-lactamase-mediated resistance mechanisms on potency of beta-lactams; and 3) in vitro activity against a panel of clinical strains producing diverse beta-lactamases. The carbapenem tebipenem had stability for many serine beta-lactamases from all molecular classes followed by cephalosporin ceftibuten. Addition of QPX7728 to tebipenem, ceftibuten and mecillinam completely reversed beta-lactamase-mediated resistance in cloned beta-lactamases from serine and metallo enzyme classes; the degree of potentiation of other beta-lactams varied according to the beta-lactamase produced. Tebipenem, ceftibuten and cefixime had the lowest MICs against laboratory strains with various combinations of beta-lactamases and the intrinsic drug-resistance mechanisms of porin and efflux mutations. There was a high degree of correlation between potency of various combinations against cloned beta-lactamases and efflux/porin mutants and the activity against clinical isolates, showing the importance of both inhibition of beta-lactamase along with minimal impact of general intrinsic resistance mechanisms affecting the beta-lactam. Tebipenem and ceftibuten appeared to be the best beta-lactam antibiotics when combined with QPX7728 for activity against Enterobacterales that produce serine or metallo beta-lactamases.

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Chromosomal -Lactamase of Klebsiella oxytoca, a New Class A Enzyme That Hydrolyzes Broad-Spectrum -Lactam Antibiotics

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.33.5.790 ◽

1989 ◽

Vol 33 (5) ◽

pp. 790-790 ◽

Cited By ~ 2

Author(s):

Y. Arakawa ◽

M. Ohta ◽

N. Kido ◽

M. Mori ◽

H. Ito ◽

...

Keyword(s):

Broad Spectrum ◽

Klebsiella Oxytoca ◽

New Class ◽

Class A

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Characterization of the penA and penR genes of Burkholderia cepacia 249 which encode the chromosomal class A penicillinase and its LysR-type transcriptional regulator.

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.41.11.2399 ◽

1997 ◽

Vol 41 (11) ◽

pp. 2399-2405 ◽

Cited By ~ 56

Author(s):

S Trépanier ◽

A Prince ◽

A Huletsky

Keyword(s):

Burkholderia Cepacia ◽

Klebsiella Oxytoca ◽

Molecular Size ◽

Beta Lactamase ◽

Beta Lactam ◽

Reading Frame ◽

Beta Lactamases ◽

Class A ◽

High Degree ◽

Degree Of Similarity

Burkholderia cepacia is recognized as an important pathogen in the lung infections of patients with cystic fibrosis. An inducible beta-lactamase activity has been associated with increased resistance to beta-lactam antibiotics in clinical isolates of B. cepacia. In this study, we report the revised sequence of the penA gene, which encodes the inducible penicillinase of B. cepacia, and show that it belongs to the molecular class A beta-lactamases and exhibits a high degree of similarity to the chromosomal beta-lactamase of Klebsiella oxytoca. Analysis of the nucleotide sequence of the DNA region directly upstream of the penA coding sequence revealed an open reading frame (penR), the transcription of which was oriented opposite to that of penA and whose initiation was 130 bp away from that of penA. Two potential ribosome-binding sites and two overlapping -10 and -35 promoter sequences were identified in the intercistronic region. The predicted translation product of penR was a polypeptide of 301 amino acids with an estimated molecular size of 33.2 kDa. The deduced polypeptide of penR showed a high degree of similarity with AmpR-like transcriptional activators of class A and C beta-lactamases, with identities of 59 and 58.7% with Pseudomonas aeruginosa PAO1 AmpR and Proteus vulgaris B317 CumR, respectively. The N-terminal portion of B. cepacia PenR was predicted to include a helix-turn-helix motif, which may bind the LysR motif identified in the intercistronic region. Induction of PenA by imipenem was shown to be dependent upon the presence of PenR. Expression of the cloned B. cepacia penA and penR genes in Escherichia coli SNO302 (ampD) resulted in a high basal and hyperinducible PenA activity. These results suggest that the regulation of the PenA penicillinase of B. cepacia 249 is similar to that observed in other class A and class C beta-lactamases that are under the control of a divergently transcribed AmpR-like regulator.

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Beta-lactamase inhibitors from laboratory to clinic.

Clinical Microbiology Reviews ◽

10.1128/cmr.1.1.109 ◽

1988 ◽

Vol 1 (1) ◽

pp. 109-123 ◽

Cited By ~ 109

Author(s):

K Bush

Keyword(s):

Side Effects ◽

Clavulanic Acid ◽

Broad Spectrum ◽

Pathogenic Bacteria ◽

Defense Mechanism ◽

Beta Lactamase ◽

Beta Lactam ◽

Beta Lactamases ◽

Beta Lactam Antibiotics ◽

Tract Infections

beta-Lactamases constitute the major defense mechanism of pathogenic bacteria against beta-lactam antibiotics. When the beta-lactam ring of this antibiotic class is hydrolyzed, antimicrobial activity is destroyed. Although beta-lactamases have been identified with clinical failures for over 40 years, enzymes with various abilities to hydrolyze specific penicillins or cephalosporins are appearing more frequently in clinical isolates. One approach to counteracting this resistance mechanism has been through the development of beta-lactamase inactivators. beta-Lactamase inhibitors include clavulanic acid and sulbactam, molecules with minimal antibiotic activity. However, when combined with safe and efficacious penicillins or cephalosporins, these inhibitors can serve to protect the familiar beta-lactam antibiotics from hydrolysis by penicillinases or broad-spectrum beta-lactamases. Both of these molecules eventually inactivate the target enzymes permanently. Although clavulanic acid exhibits more potent inhibitory activity than sulbactam, especially against the TEM-type broad-spectrum beta-lactamases, the spectrum of inhibitory activities are very similar. Neither of these inhibitors acts as a good inhibitor of the cephalosporinases. Clavulanic acid has been most frequently combined with amoxicillin in the orally active Augmentin and with ticarcillin in the parenteral beta-lactam combination Timentin. Sulbactam has been used primarily to protect ampicillin from enzymatic hydrolysis. Sulbactam has been used either in the orally absorbed prodrug form as sultamicillin or as the injectable combination ampicillin-sulbactam. Synergy has been demonstrated for these combinations for most members of the Enterobacteriaceae, although those organisms that produce cephalosporinases are not well inhibited. Synergy has also been observed for Neisseria gonorrhoeae, Haemophilus influenzae, penicillinase-producing Staphylococcus aureus, and anaerobic organisms. These antibiotic combinations have been used clinically to treat urinary tract infections, bone and soft-tissue infections, gonorrhea, respiratory infections, and otitis media. Gastrointestinal side effects have been reported for Augmentin and sultamicillin; most side effects with these agents have been mild. Although combination therapy with beta-lactamase inactivators has been used successfully, the problem of resistance development to two agents must be considered. Induction of cephalosporinases can occur with clavulanic acid. Permeability mutants could arise, especially with added pressure from a second beta-lactam.(ABSTRACT TRUNCATED AT 250 WORDS)

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Evolution of an enzyme activity: crystallographic structure at 2-A resolution of cephalosporinase from the ampC gene of Enterobacter cloacae P99 and comparison with a class A penicillinase

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.90.23.11257 ◽

1993 ◽

Vol 90 (23) ◽

pp. 11257-11261 ◽

Cited By ~ 140

Author(s):

E Lobkovsky ◽

P C Moews ◽

H Liu ◽

H Zhao ◽

J M Frere ◽

...

Keyword(s):

Binding Site ◽

Enterobacter Cloacae ◽

Crystallographic Structure ◽

Beta Lactamase ◽

Beta Lactam ◽

Beta Lactamases ◽

X Ray Crystallography ◽

Class A ◽

Beta Lactam Antibiotics ◽

Class C

The structure of the class C ampC beta-lactamase (cephalosporinase) from Enterobacter cloacae strain P99 has been established by x-ray crystallography to 2-A resolution and compared to a class A beta-lactamase (penicillinase) structure. The binding site for beta-lactam (penicillinase) structure. The binding site for beta-lactam antibiotics is generally more open than that in penicillinases, in agreement with the ability of the class C beta-lactamases to better bind third-generation cephalosporins. Four corresponding catalytic residues (Ser-64/70, Lys-67/73, Lys-315/234, and Tyr-150/Ser-130 in class C/A) lie in equivalent positions within 0.4 A. Significant differences in positions and accessibilities of Arg-349/244 may explain the inability of clavulanate-type inhibitors to effectively inactivate the class C beta-lactamases. Glu-166, required for deacylation of the beta-lactamoyl intermediate in class A penicillinases, has no counterpart in this cephalosporinase; the nearest candidate, Asp-217, is 10 A from the reactive Ser-64. A comparison of overall tertiary folding shows that the cephalosporinase, more than the penicillinase, is broadly similar to the ancestral beta-lactam-inhibited enzymes of bacterial cell wall synthesis. On this basis, it is proposed that the cephalosporinase is the older of the two beta-lactamases, and, therefore, that a local refolding in the active site, rather than a simple point mutation, was required for the primordial class C beta-lactamase to evolve to the class A beta-lactamase having an improved ability to catalyze the deacylation step of beta-lactam hydrolysis.

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839. Epidemiology of Extended-Spectrum Beta-lactamase (ESBL) Producing Enterobacteriaceae in the South East Tennessee, October-December 2017

Open Forum Infectious Diseases ◽

10.1093/ofid/ofaa439.1028 ◽

2020 ◽

Vol 7 (Supplement_1) ◽

pp. S460-S461

Author(s):

Daniel Muleta ◽

Cullen Adre ◽

Benji-Byrd Warner

Keyword(s):

Klebsiella Oxytoca ◽

Health Department ◽

Population Based ◽

Klebsiella Pneumonia ◽

Drug Resistant ◽

Beta Lactam ◽

Extended Spectrum Beta Lactamase ◽

Catchment Population ◽

Beta Lactam Antibiotics ◽

Extended Spectrum

Abstract Background The increasing spread of drug resistant gram-negative organisms is one of the major public health challenges. ESBL-producing Enterobacteriaceae has become the most common multi drug resistant pathogen in the last three decades. These organisms confer resistance to most beta-lactam antibiotics, including penicillins, third generation cephalosporins, monobactams and tazobactam. Methods The Tennessee Health Department (TDH) collaborated with CDC to pilot population based surveillance of ESBL producing organisms in Maury, Wayne, Lewis and Marshall Counties during October to December 2017. A case was defined as isolation of Escherichia coli, Klebsiella pneumoniae, or Klebsiella oxytoca resistant to at least one extended-spectrum cephalosporin (ceftazidime, cefotaxime or ceftriaxone) and non-resistant to all carbapenem antibiotics from urine or normally sterile body sites from a resident of the surveillance catchment area. A line list of ESBL-producing organisms was received from the labs that serve the catchment population. Case report forms were completed for the first ESBL culture collected from a single patient in a 30 day-period. Results A total of 154 cases were identified during the study period. E.coli constitutes 92.2% of the ESBL producing organisms followed by Klebsiella pneumonia (5.2%) and K. oxytoca (2.6%). The estimated annual incidence rate was 400.7 per 100,000 population which is more than twice of the average rates of other sites that conducted similar studies. The most common isolate source was urine (97%), and 81.2% of all cases were female. Patient ages ranged from 3-99 years, with average of 67 years. Thirty-two isolates underwent additional sequence typing and 76.7% (23) of the isolates were ST 131. 21 (91.3%) of ST-131 isolates were resistant to ciprofloxacin. Conclusion The study revealed that the incidence of ESBL producing organisms is very high in the Tennessee study area compared to other sites. The most common ESBL-producing pathogen was found to be ST 131 and most of these were resistant to ciprofloxacin suggesting that resistance to fluoroquinolone may be co-transmitted in ESBL producing pathogens through plasmids. Continued surveillance of molecular epidemiology is important to guide the prevention of the spread of drug resistant pathogens. Disclosures All Authors: No reported disclosures

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Carbapenems, a new class of beta-lactam antibiotics

The American Journal of Medicine ◽

10.1016/0002-9343(85)90097-x ◽

1985 ◽

Vol 78 (6) ◽

pp. 3-21 ◽

Cited By ~ 152

Author(s):

Jerome Birnbaum ◽

Frederick M. Kahan ◽

Helmut Kropp ◽

James S. Macdonald

Keyword(s):

Beta Lactam ◽

New Class ◽

Beta Lactam Antibiotics

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Comparison of a new cephalosporin, BMY 28142, with other broad-spectrum beta-lactam antibiotics.

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.27.2.207 ◽

1985 ◽

Vol 27 (2) ◽

pp. 207-216 ◽

Cited By ~ 73

Author(s):

R E Kessler ◽

M Bies ◽

R E Buck ◽

D R Chisholm ◽

T A Pursiano ◽

...

Keyword(s):

Broad Spectrum ◽

Beta Lactam ◽

Beta Lactam Antibiotics

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Bacterial resistance to beta-lactam antibiotics: crystal structure of beta-lactamase from Staphylococcus aureus PC1 at 2.5 A resolution

Science ◽

10.1126/science.3107125 ◽

1987 ◽

Vol 236 (4802) ◽

pp. 694-701 ◽

Cited By ~ 222

Author(s):

O Herzberg ◽

J Moult

Keyword(s):

Crystal Structure ◽

Staphylococcus Aureus ◽

Bacterial Resistance ◽

Beta Lactamase ◽

Beta Lactam ◽

Beta Lactam Antibiotics

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Growing Menace of Antibacterial Resistance in Clinical Isolates ofPseudomonas aeruginosain Nepal: An Insight of Beta-Lactamase Production

BioMed Research International ◽

10.1155/2016/6437208 ◽

2016 ◽

Vol 2016 ◽

pp. 1-8 ◽

Cited By ~ 2

Author(s):

Shamshul Ansari ◽

Rabindra Dhital ◽

Sony Shrestha ◽

Sangita Thapa ◽

Ram Puri ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Antibiotic Resistance ◽

Opportunistic Pathogen ◽

Resistance Rate ◽

Diffusion Method ◽

Beta Lactamase ◽

Beta Lactam ◽

Beta Lactam Antibiotics ◽

Microbiological Methods ◽

Immunosuppressed Patients

Introduction. Pseudomonas aeruginosais the most frequently isolated organism as it acts as the opportunistic pathogen and can cause infections in immunosuppressed patients. The production of different types of beta-lactamases renders this organism resistant to many commonly used antimicrobials. Therefore, the aim of this study was to document the antibiotic resistance rate inPseudomonas aeruginosaisolated from different clinical specimens.Methods. Pseudomonas aeruginosarecovered was identified by standard microbiological methods. Antibiotic susceptibility testing was performed by modified Kirby-Bauer disc diffusion method following Clinical and Laboratory Standard Institute (CLSI) guidelines and all the suspected isolates were tested for the production of ESBLs, MBLs, and AmpC.Results.Out of total (178) isolates, 83.1% were recovered from the inpatient department (IPD). Majority of the isolates mediated resistance towards the beta-lactam antibiotics, while nearly half of the isolates were resistant to ciprofloxacin. Most of the aminoglycosides used showed resistance rate up to 75% but amikacin proved to be better option. No resistance to polymyxin was observed. ESBLs, MBLs, and AmpC mediated resistance was seen in 33.1%, 30.9%, and 15.7% isolates, respectively.Conclusions. Antibiotic resistance rate and beta-lactamase mediated resistance were high. Thus, regular surveillance of drug resistance is of utmost importance.

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