scholarly journals Structural and biochemical characterization of the novel CTX-M-151 extended-spectrum β-lactamase and its inhibition by Avibactam

2021 ◽  
Author(s):  
Barbara Ghiglione ◽  
María Margarita Rodríguez ◽  
Florencia Brunetti ◽  
Krisztina M. Papp-Wallace ◽  
Ayumi Yoshizumi ◽  
...  
Keyword(s):  
Amino Acid ◽  
Biochemical Characterization ◽  
Sequence Divergence ◽  
Chair Conformation ◽  
Amino Acid Identity ◽  
The Novel ◽  
Proton Relay ◽  
Low Degree

The diazabicyclooctane (DBO) inhibitor, avibactam (AVI), reversibly inactivates most serine-β-lactamases including the CTX-M β-lactamases. Currently, more than 230 CTX-M unique members distributed in five clusters with less than 5% amino acid sequence divergence within each group are described. Recently, a variant named as CTX-M-151 was isolated from a Salmonella Choleraesuis strain in Japan. This variant possesses a low degree amino acid identity with the other CTX-Ms (63.2-69.7% with respect to the mature proteins), and thus it may represent a new sub-group within the family. CTX-M-151 hydrolyzes ceftriaxone better than ceftazidime (kcat/Km values 6,000-fold higher), as observed with CTX-Ms. CTX-M-151 is well inhibited by mechanism-based inhibitors like clavulanic acid (kinact/KI = 0.15 μM−1.s−1). For AVI, Ki app (0.4 μM) was comparable to KPC-2; k2/K (37,000 M−1s−1) was lower than for CTX-M-15, while the koff (0.0015 s−1) was 2-14-fold faster than other class A β-lactamases. The structure of the CTX-M-151/AVI complex (1.32 Å) reveals that AVI adopts a chair conformation with hydrogen bonds between the AVI carbamate and Ser70 and Ser237 at the oxyanion hole. Upon acylation, the side chain of Lys73 points towards Ser130 which is associated with the protonation of Glu166, supporting the role of Lys73 in the proton-relay pathway and Glu166 as the general base in deacylation. To our knowledge, this is the first chromosomally-encoded CTX-M in Salmonella Choleraesuis that shows similar hydrolytic preference towards cefotaxime/ceftriaxone when compared to ceftazidime.

Genetic and biochemical characterization of FRI-3, a novel variant of the Ambler class A carbapenemase FRI-1

2019 ◽  
Vol 74 (10) ◽  
pp. 2891-2894 ◽  
Author(s):  
Jennifer Schauer ◽  
Sören G Gatermann ◽  
Matthias Marschal ◽  
Niels Pfennigwerth
Keyword(s):  
Amino Acid ◽  
Biochemical Characterization ◽  
Amino Acid Identity ◽  
Class A ◽  
Carbapenem Resistant ◽  
New Variant ◽  
Novel Variant ◽  
Almost All ◽  
Phenotypic Tests

Abstract Objectives To characterize a new variant of the FRI class A carbapenemase isolated from an MDR clinical Enterobacter cloacae isolate. Methods A carbapenem-resistant E. cloacae was isolated from a rectal swab from a patient in an ICU in Southern Germany. Various phenotypic tests confirmed production of a putative class A carbapenemase. The new bla gene variant, blaFRI-3, and its genetic environment were characterized by WGS. Biochemical characterization was performed by heterologous expression in Escherichia coli TOP10 and by purification of the enzyme with subsequent determination of its kinetic parameters. Results PCR and sequencing carried out for different class A carbapenemase genes confirmed the presence of a novel variant of blaFRI-1. The novel variant was named FRI-3 and exhibited 91%, 96% and 92% amino acid identity to FRI-1, FRI-2 and FRI-4, respectively. E. coli TOP10 expressing blaFRI-3 showed increased resistance to almost all β-lactams. Comparing the catalytic behaviour of FRI-3 and FRI-1, it was shown that FRI-3 had the same substrate spectrum, but basically hydrolysed β-lactams less efficiently than FRI-1. WGS data revealed that blaFRI-3 was located on a 111 kb plasmid. Conclusions The biochemical characterization of FRI-3 illustrates that even a few differences in the amino acid sequence can lead to altered catalytic activities of β-lactamases belonging to the same family.

scholarly journals Molecular and Biochemical Characterization of a Novel Class A β-Lactamase (HER-1) from Escherichia hermannii

2003 ◽  
Vol 47 (8) ◽  
pp. 2669-2673 ◽  
Author(s):  
Anne Beauchef-Havard ◽  
Guillaume Arlet ◽  
Valerie Gautier ◽  
Roger Labia ◽  
Patrick Grimont ◽  
...  
Keyword(s):  
Amino Acid ◽  
Broad Spectrum ◽  
Aeromonas Hydrophila ◽  
Biochemical Characterization ◽  
Amino Acid Identity ◽  
Acid Identity ◽  
Low Level ◽  
Class A ◽  
Moderate Susceptibility

ABSTRACT Escherichia hermannii showed a low level of resistance to amoxicillin and ticarcillin, reversed by clavulanate, and a moderate susceptibility to piperacillin but was susceptible to all cephalosporins. A bla gene was cloned and encoded a typical class A β-lactamase (HER-1, pI 7.5), which shares 45, 44, 41, and 40% amino acid identity with other β-lactamases, AER-1 from Aeromonas hydrophila, MAL-1/Cko-1 from Citrobacter koseri, and TEM-1 and LEN-1, respectively. No ampR gene was detected. Only penicillins were efficiently hydrolyzed, and no hydrolysis was observed for cefuroxime and broad-spectrum cephalosporins. Sequencing of the bla gene in 12 other strains showed 98 to 100% identity with bla HER-1.

scholarly journals Molecular and Biochemical Characterization of Ambler Class A Extended-Spectrum β-Lactamase CGA-1 from Chryseobacterium gleum

2002 ◽  
Vol 46 (4) ◽  
pp. 966-970 ◽  
Author(s):  
Samuel Bellais ◽  
Thierry Naas ◽  
Patrice Nordmann
Keyword(s):  
Amino Acid ◽  
Functional Group ◽  
Susceptibility Testing ◽  
Biochemical Characterization ◽  
Amino Acid Identity ◽  
Antibiotic Susceptibility Testing ◽  
Acid Identity ◽  
Class A ◽  
Chryseobacterium Gleum

ABSTRACT Antibiotic susceptibility testing by disk diffusion of a Chryseobacterium gleum isolate, strain CIP 103039, showed a typical synergy image between clavulanic acid and expanded-spectrum cephalosporins. Shotgun cloning gave a recombinant plasmid in Escherichia coli that produced a β-lactamase, CGA-1, with a pI value of 8.9 that conferred resistance to most penicillins (except ureidopenicillins) and narrow-spectrum cephalosporins and an intermediate susceptibility to expanded-spectrum cephalosporins and aztreonam. The CGA-1 amino acid sequence shared only 60% amino acid identity with CME-1 and CME-2 from Chryseobacterium meningosepticum, the most closely related β-lactamases. CGA-1 was very likely chromosome encoded. It is a novel member of the PER subgroup of Ambler class A β-lactamases (Bush functional group 2be).

scholarly journals Biochemical Characterization of a Prokaryotic Phenylalanine Ammonia Lyase

2005 ◽  
Vol 187 (12) ◽  
pp. 4286-4289 ◽  
Author(s):  
Longkuan Xiang ◽  
Bradley S. Moore
Keyword(s):  
Amino Acid ◽  
Cinnamic Acid ◽  
Marine Bacterium ◽  
Phenylalanine Ammonia Lyase ◽  
Biochemical Characterization ◽  
The Novel ◽  
Amino Acid Residues ◽  
Primary Amino ◽  
Biochemical Features

ABSTRACT The committed biosynthetic reaction to benzoyl-coenzyme A in the marine bacterium “Streptomyces maritimus” is carried out by the novel prokaryotic phenylalanine ammonia lyase (PAL) EncP, which converts the primary amino acid l-phenylalanine to trans-cinnamic acid. Recombinant EncP is specific for l-phenylalanine and shares many biochemical features with eukaryotic PALs, which are substantially larger proteins by ∼200 amino acid residues.

scholarly journals Characterization of the Naturally Occurring Oxacillinase of Acinetobacter baumannii

2005 ◽  
Vol 49 (10) ◽  
pp. 4174-4179 ◽  
Author(s):  
Claire Héritier ◽  
Laurent Poirel ◽  
Pierre-Edouard Fournier ◽  
Jean-Michel Claverie ◽  
Didier Raoult ◽  
...  
Keyword(s):  
Amino Acid ◽  
Acinetobacter Baumannii ◽  
Biochemical Characterization ◽  
Amino Acid Identity ◽  
Acid Identity ◽  
Low Level ◽  
Narrow Spectrum ◽  
Naturally Occurring

ABSTRACT A chromosomally encoded oxacillinase, OXA-69, was characterized from Acinetobacter baumannii AYE. β-Lactamase OXA-69 shared 97% amino acid identity with the recently described OXA-51 enzyme of A. baumannii and 62 and 56% amino acid identity with the carbapenem-hydrolyzing oxacillinases OXA-24 and OXA-23, respectively. Biochemical characterization of the purified OXA-69 revealed a narrow-spectrum hydrolysis profile but including, at a low level, imipenem and meropenem. By PCR and sequencing bla OXA-69-like genes were identified in all A. baumannii strains tested (n = 12), suggesting that this oxacillinase is naturally occurring in that species.

scholarly journals Biochemical Characterization of CPS-1, a Subclass B3 Metallo-β-Lactamase from a Chryseobacterium piscium Soil Isolate

2015 ◽  
Vol 60 (3) ◽  
pp. 1869-1873 ◽  
Author(s):  
Dereje Dadi Gudeta ◽  
Simona Pollini ◽  
Jean-Denis Docquier ◽  
Valeria Bortolaia ◽  
Gian Maria Rossolini ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Broad Spectrum ◽  
Biochemical Characterization ◽  
Amino Acid Identity ◽  
Content Type ◽  
Acid Identity ◽  
Soil Isolate

CPS-1 is a subclass B3 metallo-β-lactamase from aChryseobacteriumpisciumisolate collected from soil, showing 68% amino acid identity to the GOB-1 enzyme. CPS-1 was overproduced inEscherichia coliRosetta (DE3), purified by chromatography, and biochemically characterized. This enzyme exhibits a broad-spectrum substrate profile, including penicillins, cephalosporins, and carbapenems, which overall resembles those of L1, GOB-1, and acquired subclass B3 enzymes AIM-1 and SMB-1.

Biochemical characterization of a glucoamylase from Saccharomycopsis fibuligera R64

Biologia ◽  
2011 ◽  
Vol 66 (1) ◽  
Author(s):  
Dessy Natalia ◽  
Keni Vidilaseris ◽  
Pasjan Satrimafitrah ◽  
Wangsa Ismaya ◽  
Purkan ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Amino Acid ◽  
Amino Acid Sequence Identity ◽  
Biochemical Characterization ◽  
Saccharomycopsis Fibuligera ◽  
Sequence Identity ◽  
Ic50 Value ◽  
High Amino Acid ◽  
Ph Range

AbstractGlucoamylase from the yeast Saccharomycopsis fibuligera R64 (GLL1) has successfully been purified and characterized. The molecular mass of the enzyme was 56,583 Da as determined by mass spectrometry. The purified enzyme demonstrated optimum activity in the pH range of 5.6–6.4 and at 50°C. The activity of the enzyme was inhibited by acarbose with the IC50 value of 5 μM. GLL1 shares high amino acid sequence identity with GLU1 and GLA1, which are Saccharomycopsis fibuligera glucoamylases from the strains HUT7212 and KZ, respectively. The properties of GLL1, however, resemble that of GLU1. The elucidation of the primary structure of GLL1 contributes to the explanation of this finding.

Functional and Structural Characterization of Acquired Pan-Aminoglycoside Resistance 16S rRNA Methyltransferase NpmB1

2021 ◽  
Author(s):  
Akito Kawai ◽  
Masahiro Suzuki ◽  
Kentaro Tsukamoto ◽  
Yusuke Minato ◽  
Yohei Doi
Keyword(s):  
Amino Acid ◽  
16S Rrna ◽  
Amino Acid Identity ◽  
Single Amino Acid ◽  
Aminoglycoside Resistance ◽  
E Coli ◽  
The United Kingdom ◽  
Amino Acid Variant ◽  
A Site

Post-translational methylation of the A site of 16S rRNA at position A1408 leads to pan-aminoglycoside resistance encompassing both 4,5- and 4,6-disubstituted 2-deoxystreptamine (DOS) aminoglycosides. To date, NpmA is the only acquired enzyme with such function. Here, we present function and structure of NpmB1 whose sequence was identified in Escherichia coli genomes registered from the United Kingdom. NpmB1 possesses 40% amino acid identity with NpmA1 and confers resistance to all clinically relevant aminoglycosides including 4,5-DOS agents. Phylogenetic analysis of NpmB1 and NpmB2, its single amino acid variant, revealed that the encoding gene was likely acquired by E. coli from a soil bacterium. The structure of NpmB1 suggests that it requires a structural change of the β6/7 linker in order to bind to 16S rRNA. These findings establish NpmB1 and NpmB2 as the second group of acquired pan-aminoglycoside resistance 16S rRNA methyltransferases.

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