The bla gene of the cephamycin cluster of Streptomyces clavuligerus encodes a class A beta-lactamase of low enzymatic activity.

Four beta-lactamases excreted by Gram-positive bacteria exhibited microheterogeneity when analysed by chromatofocusing or ion-exchange chromatography. Ragged N-termini were in part responsible for the charge variants, but deamidation of an asparagine residue was also involved, at least for the Bacillus licheniformis enzyme. The activity of a contaminating proteinase could also be demonstrated in the case of Actinomadura R39 beta-lactamase. With that enzyme, proteolysis resulted in partial inactivation, but the inactivated fragments were easily separated from the active forms. With these, as with the other enzymes, the kinetic parameters of the major variants were identical with those of the mixture within the limits of experimental error, so that the catalytic properties of these enzymes can be determined with the ‘heterogeneous’ preparations.

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Characterization of beta-lactamase gene blaPER-2, which encodes an extended-spectrum class A beta-lactamase.

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.40.3.616 ◽

1996 ◽

Vol 40 (3) ◽

pp. 616-620 ◽

Cited By ~ 82

Author(s):

A Bauernfeind ◽

I Stemplinger ◽

R Jungwirth ◽

P Mangold ◽

S Amann ◽

...

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Beta Lactamase ◽

Reading Frame ◽

Beta Lactamases ◽

Class A ◽

Extended Spectrum ◽

The Family ◽

Extended Spectrum Beta Lactamases ◽

Lactamase Gene

Plasmidic extended-spectrum beta-lactamases of Ambler class A are mostly inactive against ceftibuten. Salmonella typhimurium JMC isolated in Argentina harbors a bla gene located on a plasmid (pMVP-5) which confers transferable resistance to oxyiminocephalosporins, aztreonam, and ceftibuten. The beta-lactamase PER-2 (formerly ceftibutenase-1; CTI-1) is highly susceptible to inhibition by clavulanate and is located at a pI of 5.4 after isoelectric focusing. The blaPER-2 gene was cloned and sequenced. The nucleotide sequence of a 2.2-kb insert in vector pBluescript includes an open reading frame of 927 bp. Comparison of the deduced amino acid sequence of PER-2 with those of other beta-lactamases indicates that PER-2 is not closely related to TEM or SHV enzymes (25 to 26% homology). PER-2 is most closely related to PER-1 (86.4% homology), an Ambler class A enzyme first detected in Pseudomonas aeruginosa. An enzyme with an amino acid sequence identical to that of PER-1, meanwhile, was found in various members of the family Enterobacteriaceae isolated from patients in Turkey. Our data indicate that PER-2 and PER-1 represent a new group of Ambler class A extended-spectrum beta-lactamases. PER-2 so far has been detected only in pathogens (S. typhimurium, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis) isolated from patients in South America, while the incidence of PER-1-producing strains so far has been restricted to Turkey, where it occurs both in members of the family Enterobacteriaceae and in P. aeruginosa.

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In vivo functional analysis of a class A β-lactamase-related protein essential for clavulanic acid biosynthesis in Streptomyces clavuligerus

PLoS ONE ◽

10.1371/journal.pone.0215960 ◽

2019 ◽

Vol 14 (4) ◽

pp. e0215960 ◽

Cited By ~ 2

Author(s):

Santosh K. Srivastava ◽

Kelcey S. King ◽

Nader F. AbuSara ◽

Chelsea J. Malayny ◽

Brandon M. Piercey ◽

...

Keyword(s):

Functional Analysis ◽

Clavulanic Acid ◽

Streptomyces Clavuligerus ◽

Related Protein ◽

Acid Biosynthesis ◽

Class A

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4,7-Dichloro benzothien-2-yl sulfonylaminomethyl boronic acid: First boronic acid-derived beta-lactamase inhibitor with class A, C, and D activity

Bioorganic & Medicinal Chemistry Letters ◽

10.1016/j.bmcl.2010.02.065 ◽

2010 ◽

Vol 20 (8) ◽

pp. 2622-2624 ◽

Cited By ~ 18

Author(s):

Qiang Tan ◽

Aimie M. Ogawa ◽

Ronald E. Painter ◽

Young-Whan Park ◽

Katherine Young ◽

...

Keyword(s):

Boronic Acid ◽

Beta Lactamase ◽

Class A ◽

Lactamase Inhibitor

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Sequence analysis and enzyme kinetics of the L2 serine beta-lactamase from Stenotrophomonas maltophilia.

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.41.7.1460 ◽

1997 ◽

Vol 41 (7) ◽

pp. 1460-1464 ◽

Cited By ~ 74

Author(s):

T R Walsh ◽

A P MacGowan ◽

P M Bennett

Keyword(s):

Amino Acids ◽

Sequence Analysis ◽

Clavulanic Acid ◽

Active Site ◽

Stenotrophomonas Maltophilia ◽

Leader Peptide ◽

Beta Lactamase ◽

Class A ◽

Cloned Gene ◽

Kinetics Of

The L2 serine active-site beta-lactamase from Stenotrophomonas maltophilia has been classified as a clavulanic acid-sensitive cephalosporinase. The gene encoding this enzyme from S. maltophilia 1275 IID has been cloned on a 3.3-kb fragment into pK18 under the control of a Ptac promoter to generate recombinant plasmid pUB5840; when expressed in Escherichia coli, this gene confers resistance to cephalosporins and penicillins. Sequence analysis has revealed an open reading frame (ORF) of 909 bp with a GC content of 71.6%, comparable to that of the L1 metallo-beta-lactamase gene (68.4%) from the same bacterium. The ORF encodes an unmodified protein of 303 amino acids with a predicted molecular mass of 31.5 kDa, accommodating a putative leader peptide of 27 amino acids. Comparison of the amino acid sequence with those of other beta-lactamases showed it to be most closely related (54% identity) to the BLA-A beta-lactamase from Yersinia enterocolitica. Sequence identity is most obvious near the STXK active-site motif and the SDN loop motif common to all serine active-site penicillinases. Sequences outside the conserved regions display low homology with comparable regions of other class A penicillinases. Kinetics of the enzyme from the cloned gene demonstrated an increase in activity with cefotaxime but markedly less activity with imipenem than previously reported. Hence, the S. maltophilia L2 beta-lactamase is an inducible Ambler class A beta-lactamase which would account for the sensitivity to clavulanic acid.

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The K1 β-lactamase of Klebsiella pneumoniae

Biochemical Journal ◽

10.1042/bj2430561 ◽

1987 ◽

Vol 243 (2) ◽

pp. 561-567 ◽

Cited By ~ 12

Author(s):

B Joris ◽

F De Meester ◽

M Galleni ◽

J M Frère ◽

J Van Beeumen

Keyword(s):

Klebsiella Pneumoniae ◽

Active Site ◽

Cysteine Residue ◽

Klebsiella Aerogenes ◽

Beta Lactamase ◽

Serine Residue ◽

Class A

beta-Lactamase K1 was purified from Klebsiella pneumoniae SC10436. It is very similar to the enzyme produced by Klebsiella aerogenes 1082E and described by Emanuel, Gagnon & Waley [Biochem. J. (1986) 234, 343-347]. An active-site peptide was isolated after labelling of the enzyme with tritiated beta-iodopenicillanate. A cysteine residue was found just before the active-site serine residue. This result could explain the properties of the enzyme after modification by thiol-blocking reagents. The sequence of the active-site peptide clearly established the enzyme as a class A beta-lactamase.

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Imipenem as substrate and inhibitor of β-lactamases

Biochemical Journal ◽

10.1042/bj2530323 ◽

1988 ◽

Vol 253 (2) ◽

pp. 323-328 ◽

Cited By ~ 18

Author(s):

J Monks ◽

S G Waley

Keyword(s):

Pseudomonas Aeruginosa ◽

Bacillus Cereus ◽

Reversible Reaction ◽

Clinical Use ◽

Beta Lactamase ◽

Beta Lactamases ◽

Class A ◽

Class C ◽

Kinetics Of ◽

Carbapenem Antibiotic

The interaction between imipenem, a carbapenem antibiotic, and two representative beta-lactamases has been studied. The first enzyme was beta-lactamase I, a class-A beta-lactamase from Bacillus cereus; imipenem behaved as a slow substrate (kcat. 6.7 min-1, Km 0.4 mM at 30 degrees C and at pH 7) that reacted by a branched pathway. There was transient formation of an altered species formed in a reversible reaction; this species was probably an acyl-enzyme in a slightly altered, but considerably more labile, conformation. The kinetics of the reaction were investigated by measuring both the concentration of the substrate and the activity of the enzyme, which fell and then rose again more slowly. The second enzyme was the chromosomal class-C beta-lactamase from Pseudomonas aeruginosa; imipenem was a substrate with a low kcat. (0.8 min-1) and a low Km (0.7 microM). Possible implications for the clinical use of imipenem are considered.

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Identification of a metagenomic gene cluster containing a new class A beta‐lactamase and toxin‐antitoxin systems

MicrobiologyOpen ◽

10.1002/mbo3.104 ◽

2013 ◽

Vol 2 (4) ◽

pp. 674-683 ◽

Cited By ~ 5

Author(s):

Ken Vercammen ◽

Tamara Garcia‐Armisen ◽

Nathalie Goeders ◽

Laurence Melderen ◽

Josselin Bodilis ◽

...

Keyword(s):

Gene Cluster ◽

Beta Lactamase ◽

New Class ◽

Class A

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Phosphonate monoester inhibitors of class A β-lactamases

Biochemical Journal ◽

10.1042/bj2750793 ◽

1991 ◽

Vol 275 (3) ◽

pp. 793-795 ◽

Cited By ~ 27

Author(s):

J Rahil ◽

R F Pratt

Keyword(s):

Enzyme Inhibition ◽

Mechanism Of Action ◽

Active Site ◽

General Structure ◽

Beta Lactamase ◽

Beta Lactamases ◽

Inhibitory Ability ◽

Class A ◽

Structure Formula ◽

Slow Turnover

Phosphonate monoesters with the general structure: [formula: see text] are inhibitors of representative class A and class C beta-lactamases. This result extends the range of this type of inhibitor to the class A enzymes. Compounds where X is an electron-withdrawing substituent are better inhibitors than the unsubstituted analogue (X = H), and enzyme inhibition is concerted with stoichiometric release of the substituted phenol. Slow turnover of the phosphonates also occurs. These observations support the proposition that the mechanism of action of these inhibitors involves phosphorylation of the beta-lactamase active site. The inhibitory ability of these phosphonates suggests that the beta-lactamase active site is very effective at stabilizing negatively charged transition states. One of the compounds described also inactivated the Streptomyces R61 D-alanyl-D-alanine carboxypeptidase/transpeptidase.

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